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    18 April 2021, Volume 25 Issue 11 Previous Issue    Next Issue
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    Interaction proteomics of long noncoding RNA uc431+ gene in postmenopausal osteoporosis
    Li Shengqiang, Xie Bingying, Chen Juan, Xie Lihua, Huang Jingwen, Ge Jirong
    2021, 25 (11):  1641-1646.  doi: 10.3969/j.issn.2095-4344.3107
    Abstract ( 78 )   PDF (1308KB) ( 18 )   Save
    BACKGROUND: Overexpression of LncRNA TUG1 promotes osteoclast proliferation and inhibits apoptosis. Knockdown with siRNA has achieved the opposite result, indicating that knockdown of LncRNA TUG1 may become a potential target for osteoporosis treatment.
    OBJECTIVE: To study the proteins interacting with the non-coding RNA uc431+ associated with postmenopausal osteoporosis and carry out bioinformatics analysis. 
    METHODS: Human monocytic leukemia cell line (THP-1) cells crosslinked with formaldehyde were broken by ultrasound. The experimental group was hybridized with magnetic beads combined with biotin labeled specific probes. The control group was hybridized with the magnetic beads of non-specific probes. The obtained peptides were identified using mass spectrometry. The data were searched and quantified by MaxQuant. The quantitative results of the two sets of samples were statistically analyzed, and the corresponding enrichment proteins were obtained. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, protein-protein interaction analysis and display were performed, and the protein-protein interaction network was constructed. 
    RESULTS AND CONCLUSION: The total number of peptides obtained was 918, with 271 proteins in total, and the total number of proteins after filtration was 241. Compared with the control group, there were 10 differential proteins in the experimental group, including DDOST, DMBT1, HPD, IGLL5, IGK, LTF, LYZ, MUC5AC, PIGR, and RPL23. GO enrichment analysis showed that they were involved in biological processes such as defense reaction, leukocyte activation, ribosomal rRNA binding, lysozyme activity and other molecular functions. KEGG pathway analysis predicted that they were involved in ubiquinone and other terpenoid-quinone biosynthesis, phenylalanine metabolism, ribosome and salivary secretion. Combined with the analysis of proteomics and bioinformatics, it is predicted that uc431+, a gene related to postmenopausal osteoporosis, may be involved in immune regulation and bone metabolism by interacting proteins.
    Key words: osteoporosis; menopause; non-coding RNA; proteomics; mass spectrum; lactoferrin; bone metabolism; bioinformatics

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    Clinical application of enhanced recovery after surgery in the perioperative period of anterior cruciate ligament reconstruction
    Sun Kai, Chen Lei, Mai Yao, Hu Hua, Chen Liang, Zhong Jun, Hu Yong, Qiu Bo
    2021, 25 (11):  1647-1651.  doi: 10.3969/j.issn.2095-4344.3086
    Abstract ( 106 )   PDF (665KB) ( 19 )   Save
    BACKGROUND: Anterior cruciate ligament injury, with an increasing incidence in recent years, will seriously impact the motor function of the knee joint. Rehabilitation exercise after anterior cruciate ligament reconstruction is directly related to whether the patient can obtain ideal joint function. It is of important clinical significance to introduce the concept of enhanced recovery after surgery (ERAS) into the perioperative management of arthroscopic anterior cruciate ligament reconstruction.
    OBJECTIVE: To explore the clinical effect of ERAS in the perioperative period of anterior cruciate ligament reconstruction. 
    METHODS: Sixty patients with anterior cruciate ligament injury, who were treated in the Department of Orthopedics, Renmin Hospital of Wuhan University from July 2016 to July 2018, were randomly divided into ERAS group (n=30) and traditional treatment group (n=30). Postoperative Visual Analogue Scale score, recovery time of knee range of motion, Lysholm knee scoring scale score, hospitalization duration, patients’ satisfaction rating were compared between the two groups. The implementation of the study complied with the Declaration of Helsinki and the relevant ethical requirements of Wuhan University Renmin Hospital, and all the enrolled patients were fully informed of the trial process.
    RESULTS AND CONCLUSION: The Visual Analogue Scale scores in the ERAS group were significantly lower than those in the traditional treatment group at 1, 24 hours, 2, 3, and 7 days after surgery (P < 0.05). The time that the range of motion returned to 30°, 60°, 90° and 120° after surgery in the ERAS group was significantly less than that in the traditional treatment group (P < 0.05). The Lysholm knee scoring scale scores in 3 months and 6 months after surgery in the ERAS group were significantly higher than those in the traditional treatment group (P < 0.05). The length of hospitalization in the ERAS group was significantly lower than that in the traditional treatment group (P < 0.05). The patients’ satisfaction rating in the ERAS group was significantly higher than that in the traditional treatment group (P < 0.05). To conclude, the introduction of ERAS concept into the perioperative management of anterior cruciate ligament reconstruction has obvious merits in reducing anxiety, relieving postoperative pain, reducing postoperative complications, promoting early recovery of patients and accelerating the recovery of knee joint function.

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    Mechanism of DAIa2GIP inhibiting mitochondrial apoptosis in chondrocytes
    Wang Yue, Wang Xinjun, Yuan Yinpeng, Wang Yuze
    2021, 25 (11):  1652-1657.  doi: 10.3969/j.issn.2095-4344.3093
    Abstract ( 71 )   PDF (1089KB) ( 18 )   Save
    BACKGROUND: The previous research of our group has shown that DAIa2GIP, a new analogue of glucose dependent insulin stimulating polypeptide, has a protective effect on chondrocytes, but its mechanism is not clear.
    OBJECTIVE: To observe the effect of DAIa2GIP on interleukin-1β (IL-1β) induced apoptosis of chondrocytes, and to explore its molecular biological mechanism.
    METHODS: The costal chondrocytes of Sprague-Dawley rats were extracted and identified by SABC immunohistochemistry. The third generation cells were divided into six groups: (1) normal control group; (2) IL-1β induction group; (3) DAIa2GIP+IL-1β induction group; (4) DAIa2GIP+IL-1β+pro3GIP (GIPR antagonists) induction group; (5) DAIa2GIP induction group; (6) pro3GIP induction group. After 48 hours of drug treatment, mitochondrial membrane potential was tested, cell apoptosis was detected using Annexin-V FITC Kit (phosphatidylserine eversion analysis), calcium overload of cells was determined under laser confocal microscope, and cytochrome C content were measured by ELISA. The study protocol was approved by the Animal Ethics Committee of Shanxi Medical University. 
    RESULTS AND CONCLUSION: SABC immunohistochemical staining showed that collagen II could be developed as chondrocytes; the maintenance degree of mitochondrial membrane potential was significantly higher in the IL-1β+DAIa2GIP incubation group than the IL-1β induction group, but the potential in both groups was lower than that in the normal control group. The apoptotic rate of chondrocytes in the IL-1β+DAIa2GIP incubation group was significantly lower than that in the IL-1β induction group, but the apoptotic rate in both groups was higher than that in the normal control group. The degree of calcium overload was compared with fluorescence intensity, and the result showed that the IL-1β+DAIa2GIP incubation group had a significantly higher intensity than the IL-1β induction group, and the fluorescence intensity in both groups was higher than that in the normal control group. The release of chondrocyte mitochondrial cytochrome C in the IL-1β+DAIa2GIP incubation group was significantly lower than that in the IL-1β induction group (P < 0.01), and the release amount in both groups was significantly higher than that in the normal control group (P < 0.01). The above four indicators showed no significant difference between the IL-1β induction group and the DAIa2GIP+IL-1β+pro3GIP group, but the values in both groups were lower than those in the control group. There was no significant difference among the normal control group, DAIa2GIP induction group, and pro3GIP induction group. To conclude, DALa2GIP can effectively antagonize IL-1β induced mitochondrial dysfunction of rat chondrocytes, thus antagonizing chondrocyte apoptosis. In this process, DALa2GIP can effectively reduce the degree of calcium overload and the release of cytochrome C.

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    Hyperbaric oxygen alleviates scar adhesion at the base and shortens the time of revascularization of delayed skin flap in rats
    Hu Yigao, Li Gaofeng
    2021, 25 (11):  1658-1663.  doi: 10.3969/j.issn.2095-4344.3085
    Abstract ( 62 )   PDF (1574KB) ( 22 )   Save
    BACKGROUND: Hyperbaric oxygen can improve the survival rate of the flap, but whether hyperbaric oxygen therapy can reduce the scar adhesion between the delayed flap and the base, and shorten the time of delayed flap revascularization have not been documented.
    OBJECTIVE: To observe the changes of the delayed skin flap and the scar tissue at the base on the back of the rat after hyperbaric oxygen therapy, and to seek a safer and more time-saving method of delayed flap formation.
    METHODS: This experiment simulated the delayed flap model of the human reconstructed ear on the back of Sprague-Dawley rats. Healthy male Sprague-Dawley rats were randomly divided into two groups: a control group and a hyperbaric oxygen treatment group. In the latter group, the rats were placed in the infant hyperbaric oxygen chamber for hyperbaric oxygen treatment for 5 days after surgery. The blood flow of the flap was recorded preoperatively and daily after surgery. Scar tissue thickness, type I/III collagen ratio, vascular number, vascular diameter and the number of vascular endothelial growth factor-positive cells in the flap were recorded preoperatively and on the 1st, 7th, 14th and 21st days postoperatively. 
    RESULTS AND CONCLUSION: The blood flow of the flaps in both groups decreased significantly on the 1st postoperative day , and then began to increase. The blood flow in the hyperbaric oxygen treatment group was significantly higher than that in the control group on the 1st, 7th, 14th and 21st days postoperatively (P < 0.05). The subcutaneous tissue edema in the hyperbaric oxygen treatment group was slighter than that in the control group, and the tissue inflammatory cell infiltration was less. On the 21st postoperatively day, the thickness of scar tissue adhesion at the base in the hyperbaric oxygen treatment group was smaller than that in the control group (P < 0.05). The ratio of type I/III collagen in the hyperbaric oxygen treatment group was lower than that in the control group (P < 0.05). On the 7th, 14th, and 21st days postoperatively, the number of blood vessels in the hyperbaric oxygen treatment group was higher than that in the control group (P < 0.05). The vascular diameter in the two groups was the smallest on the 1st postoperative day, and the largest on the 14th postoperative day. The vascular diameter in the hyperbaric oxygen treatment group was significantly larger than that in control group on the 1st postoperative day (P < 0.05). The number of vascular endothelial growth factor positive cells in both groups began to increase on the 1st postoperativel day, and reached the maximum value on the 14th postoperative day. The number of vascular endothelial growth factor positive cells in the hyperbaric oxygen treatment group was higher than that in the control group at each observation time point (P < 0.05). To conclude, hyperbaric oxygen can reduce the scar adhesion between delayed skin flap and the basement and increase the blood flow of the delayed flaps in rats. Its mechanism may be related to the promotion of revascularization.

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    Salidroside inhibits apoptosis of retinal Müller cells induced by high glucose in rats
    Zhao Ning, Yu Hongdan, Feng Zhen, Ding Jiayuan, Liu Xuezheng
    2021, 25 (11):  1664-1669.  doi: 10.3969/j.issn.2095-4344.3108
    Abstract ( 74 )   PDF (863KB) ( 21 )   Save
    BACKGROUND: It has been found that salidroside can improve diabetic retinopathy. However, it is not clear whether salidroside can protect retinal Müller cells (rMC-1) against damage induced by high glucose.
    OBJECTIVE: To investigate the effect and mechanism of salidroside on oxidative stress and apoptosis induced by high glucose in rat rMC-1 cells.  
    METHODS: The rMC-1 cells were divided into four groups: normal control group, high glucose group (culture medium with a final glucose concentration of 35.5 mmol/L), salidroside group (treatment with salidroside for 4 hours followed by co-culture with high glucose) and PI3K inhibitor  group (treatment with salidroside and PI3K inhibitor for 4 hours followed by co-culture with high glucose). The viability of the rMC-1 cells was measured by cell counting kit-8 assay. Reactive oxygen species production was detected by DCFH-DA. The superoxide dismutase and catalase activities were tested by specific kits. Cell apoptosis was detected by flow cytometry. Western blot was used to detect the expression levels of Cleaved caspase-3, PI3K, AKT and phosphorylated AKT. 
    RESULTS AND CONCLUSION: Compared with the normal control group , the cell viability and the activities of antioxidant enzymes (superoxide dismutase, catalase) in the high glucose group were significantly decreased, the production of reactive oxygen species, apoptotic rate and the level of Cleaved caspase-3 were significantly increased, and the phosphorylated AKT/AKT ratio was down-regulated (P < 0.05). Compared with the high glucose group, salidroside significantly enhanced the cell viability and increased the activities of antioxidant enzymes (superoxide dismutase, catalase), decreased the production of reactive oxygen species, reduced the apoptotic rate of the cells and downregulated the expression level of Cleaved caspase-3 (P < 0.05). Salidroside also activated the phosphorylation of Akt (P < 0.05). Whereas, the addition of LY294002, a pharmacological inhibitor of PI3K, showed similar results in the high glucose group (P > 0.05). To conclude, salidroside can protect rMC-1 cells through inhibiting apoptosis and oxidative stress induced by high glucose. The main mechanism responsible for the inhibition of oxidative stress is the activation of the PI3K/Akt pathway.

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    Ultrasound-guided injection of hyaluronic acid and corticosteroid for treating plantar fasciitis: evaluation of pain, fascia thickness and ankle-foot function
    Zhang Yicen, Wang Peixin, Liu Zhicheng
    2021, 25 (11):  1670-1674.  doi: 10.3969/j.issn.2095-4344.3040
    Abstract ( 126 )   PDF (621KB) ( 35 )   Save
    BACKGROUND: Sodium hyaluronate injection has been used in many musculoskeletal diseases, including knee osteoarthritis, frozen shoulder, rheumatoid arthritis, and other tendon and soft tissue pain.
    OBJECTIVE: To evaluate the effect of sodium hyaluronate injection and corticosteroid injection in reducing the symptoms of plantar fasciitis.
    METHODS: Sixty-five patients with plantar fasciitis were randomly divided into two groups: sodium hyaluronate group (32 cases) and corticosteroid group (33 cases). Two groups of patients were injected with 2.5 mL of sodium hyaluronate (25 mg/2.5 mL) and compound betamethasone (5 mg of diprosone and 2 mg of betamethasone sodium phosphate) under the guidance of ultrasound, respectively. The treatment interval was 7 days, twice in total. The Visual Analogue Scale (VAS) score, plantar fascia thickness and the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score were measured before treatment, 1 month and 3 months after treatment.
    RESULTS AND CONCLUSION: At 1 month after treatment, compared with the baseline data, the VAS and plantar fascia thickness of the two groups decreased, and the AOFAS ankle-hindfoot score increased (P < 0.05). Moreover, the VAS score of the corticosteroid group was significantly lower than that of the sodium hyaluronate group (P < 0.05). At 3 months after treatment, compared with the data at 1 month after treatment, the treatment effect was significantly improved in both two groups (P < 0.05), and there was no significant difference between the two groups (P > 0.05). To conclude, corticosteroids and sodium hyaluronate are both effective to treat plantar fasciitis, but the effect of corticosteroids is better within 1 month. To avoid the potential risk of corticosteroids, sodium hyaluronate can be a substitute of corticosteroids.

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    Aerobic exercise protects the rat brain against senile dementia induced by amyloid beta protein 1-42
    Cao Haixin, Wang Xiaomei
    2021, 25 (11):  1675-1681.  doi: 10.3969/j.issn.2095-4344.3106
    Abstract ( 68 )   PDF (1240KB) ( 24 )   Save
    BACKGROUND: Aerobic exercise has been experimentally proved to improve the blood supply of the brain after brain injury. In clinical practice, aerobic exercise is effective in treating Alzheimer’s disease, but the research on its mechanism is still at a blank stage.
    OBJECTIVE: To explore the mechanism by which aerobic exercise protects the brain of amyloid β-protein (Aβ) 1-42-induced dementia rats through the adenosine monophosphate activated protein kinase (AMPK)/endothelial nitric oxide synthase (eNOS)/nuclear factor κB (NF-κB) signaling pathway.
    METHODS: A total of 80 SPF Sprague-Dawley male rats were randomly divided into 4 groups (n=20 per group): sham operation group, model group, aerobic exercise group, and inhibitor group. Animal model of Aβ1-42-induced Alzheimer’s disease was made in each rat except for the sham operation group. In the sham operation group, 2 μL of normal saline was injected after cutting the dura mater. On the 3rd day after modeling, the inhibitor group was intraperitoneally injected with 2 mL of 50 mg/L AMPK Compound C, and the other groups were injected with the same amount of normal saline. After the injection, the rats in the aerobic exercise group and inhibitor group received 12 weeks of aerobic exercise training, and those in the sham operation group and model group received no treatment. After exercise, the changes in the spatial learning and memory ability of the rats were detected by water maze test and step down test; laser speckle imaging, hematoxylin-eosin staining, ELISA, and immunofluorescence were used to detect the changes in cerebral cortex blood flow, cerebral infarction volume, neuronal damage in brain tissue, expression levels of interleukin 1β, interleukin 6, and tumor necrosis factor α in brain tissue, nuclear translocation of NF-κB; western blot assay was used to detect the expression levels of p-AMPK, p-eNOS and NF-κB proteins. 
    RESULTS AND CONCLUSION: Compared with the sham operation group, the rats in the model group had significantly prolonged escape latency, reduced number of times passing through the original quadrant where the original platform was located, reduced blood perfusion volume in the cerebral cortex, increased expression levels of interleukin 6 and tumor necrosis factor α, enhanced fluorescence intensity of nuclear translocation of NF-κB in brain tissue, reduced expression levels of p-AMPK and p-eNOS in brain tissue, and increased expression of NF-κB protein (all P < 0.05). Compared with the model group, in the aerobic exercise group, the escape latency was significantly decreased, the number of times passing through the quadrant where the original platform was located was significantly increased, the cerebral cortex blood perfusion was significantly increased, the apoptosis rate in the brain tissue and expression levels of interleukin 1β, interleukin 6 and tumor necrosis factor α decreased, the fluorescence intensity of NF-κB nuclear translocation in the brain tissue was significantly weakened, and the expression levels of p-AMPK and p-eNOS in the brain tissue were significantly increased, and NF-κB protein expression was significantly decreased (all P < 0.05). Compared with the aerobic exercise group, the above indicators in the inhibitor group were significantly reversed (P < 0.05). To conclude, aerobic exercise may inhibit the activation of NF-κB by activating AMPK/eNOS/NF-κB signals, and thereby reduce inflammatory response, which plays a protective effect against Aβ1-42 induced senile dementia.

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    Identification and survival analysis of tTA/tetO-CCKR-2 double transgenic mice
    Gao Ziqing, Ruan Sibei, Li Li, Ling Feng, Tang Xiaoqin, Kang Qingmei, Luo Siyi, Luo Jing, Tang Yaping, Tang Mingxi
    2021, 25 (11):  1682-1687.  doi: 10.3969/j.issn.2095-4344.3079
    Abstract ( 73 )   PDF (873KB) ( 12 )   Save
    BACKGROUND: The inducible forebrain-specific cholecystokinin receptor-2 (CCKR-2) double transgenic (tTA/tetO-CCKR-2 tg, abbreviated as dtg) mice are an ideal model of anxiety-related diseases. However, there is still a lack of model identification and life related data
    OBJECTIVE: To identify the genomic DNA of the offspring and the specific expression of CCKR-2 transgene in the forebrain, and to analyze the survival probability of dtg mice. 
    METHODS: α-CaMKII/tTA single transgenic mice and tetO-CCKR-2 single transgenic mice were cross-fertilized to construct a dtg mouse model. The genomic DNA was extracted from the tail of the offspring, and the genotypes were detected by PCR and agarose gel electrophoresis. Wild-type (WT) mice were used as controls. In situ hybridization was used to detect the expression of CCKR-2. Survival of dtg mice and WT mice (30 females and 30 males) was observed and recorded within 2 years. The study protocol was approved by the Experimental Animal Ethics Committee of Southwest Medical University, with an approval No. 20150068.
    RESULTS AND CONCLUSION: Agarose gel electrophoresis results showed the molecular weight of the PCR products of dtg mice was consistent with the expected target gene fragment. In situ hybridization results showed a strong signal of CCKR-2 was detected in the forebrain of dtg mice, but hardly present in the WT mice. The median survival time of dtg mice was 76 weeks in females and 77 weeks in males. The survival probability was decreased with age in dtg mice. The survival probability of WT mice was significantly better than that of dtg mice (P < 0.001). There was no significant sex difference between males and females of dtg mice (P=0.577). Therefore, the specific expression of CCKR-2 transgene in the forebrain can be identified using PCR amplification, genomic DNA extraction, agarose gel electrophoresis, and in situ hybridization. tTA/tetO-CCKR-2 double transgenic induction may shorten the survival time of mice, but no significant difference is observed between the females and males of dtg mice.
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    A Guangxi Bama minipig model of Trimeresurus stejnegeri snakebite: modeling and evaluation
    Mo Caifeng, Cheng Xiaoyang, Liao Ming, He Dongling, Huang Zhi
    2021, 25 (11):  1688-1692.  doi: 10.3969/j.issn.2095-4344.3087
    Abstract ( 74 )   PDF (1070KB) ( 30 )   Save
    BACKGROUND:  At present, there are not many research methods on the pathological mechanism of Trimeresurus stejnegeri snakebite, and there are few methods for constructing animal models.
    OBJECTIVE: To establish and evaluate the Guangxi Bama minipig model for research on the pathological mechanism, diagnosis and treatment of Trimeresurus stejnegeri snakebite. 
    METHODS: Based on the 50% lethal dose of intramuscularly injected Trimeresurus stejnegeri venom to mice, the theoretical 50% lethal venom dose for Bama minipigs was calculated by the equivalent dose coefficient conversion and reduction algorithm, and the body surface area conversion algorithm. Twelve Bama minipigs were randomly divided into a normal group (n=6) and a model group (n=6). The model group was injected intramuscularly with 0.2 mL/kg snake venom 1/3 of the theoretical 50% lethal dose (0.643 mg/kg). The control group was injected with the same amount of normal saline. After snake venom injection, the poisoning symptoms of Bama minipigs were observed. Two groups of animal blood samples were collected before, 6 hours and 24 hours after snake venom injection. Blood routine test, four coagulation items, blood biochemistry and electrolyte were detected. Histopathological changes of the heart, brain, lung, liver, and kidney as well as the injection site were observed by hematoxylin-eosin staining. The study protocol was approved by the Laboratory Animal Ethics Committee (approval No. 201909013). 
    RESULTS AND CONCLUSION: There was swelling at the wound of the piglet accompanied with blood blisters after snake venom injection. The affected limbs and the surrounding area were swollen and spread rapidly to the proximal end. The piglets walked all the time because of the pain, and no animal died during the experiment. Compared with the control group pig, the model group had higher red blood cell count, white blood cell count, alanine aminotransferase, D-dimer count, longer prothrombin time, and lower fibrinogen and platelet count. Hematoxylin-eosin staining showed that, compared with the control group, capillary permeability of piglet lung tissues in the model group was increased with hyperemia and edema. Edema, bleeding, degeneration and necrosis were seen in the muscle tissue on the injection site. No obvious abnormalities in other organs and tissues were observed. To conclude, this method can be used to establish a pig model of Trimeresurus stejnegeri snakebite that can reflect the pathophysiological process of Trimeresurus stejnegeri snakebite. It has operability and repeatability that can be used to study the pathophysiological mechanism of Trimeresurus stejnegeri snakebite.

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    Effects of aerobic and resistance exercises on endoplasmic reticulum stress-related proteins in diabetic peripheral neuropathy rats
    Shui Xiaoping, Li Chunying, Cao Yanxia, Su Quansheng
    2021, 25 (11):  1693-1698.  doi: 10.3969/j.issn.2095-4344.3075
    Abstract ( 71 )   PDF (860KB) ( 15 )   Save
    BACKGROUND: Studies have suggested that endoplasmic reticulum stress is involved in the process of diabetic peripheral neuropathy, and improving the state of endoplasmic reticulum stress can reduce the degree of diabetic peripheral neuropathy. Whether exercise has a positive effect on endoplasmic reticulum stress in diabetic peripheral nerves is rarely documented.
    OBJECTIVE: To observe the effects of aerobic and resistance exercise on the expression of Grp78, Chop and Caspase-12 in the endoplasmic reticulum stress signal pathway in the sciatic nerve of type 2 diabetic mellitus (T2DM) rats. 
    METHODS: Sixty Sprague-Dawley male rats were divided into normal control group (normal diet, n=24) and T2DM model group (n=36). Rats in the T2DM model group were fed with high-fat and high-sugar diet for 8 weeks to establish T2DM models. Rats in the normal control group were randomly subdivided into quiet control group, aerobic exercise group and resistance exercise group, while those in the T2DM model group were randomized into quiet T2DM group, T2DM aerobic exercise group, and T2DM resistance exercise group. Aerobic and resistance exercises in corresponding groups lasted for 8 weeks. After 8 weeks exercise, fasting blood glucose, insulin level, insulin resistance index, mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured. The structural changes of the sciatic nerve were observed by transmission electron microscope. The expression levels of Grp78, Chop and Caspase-12 in the sciatic nerve were detected by western blot. An ethic approval was obtained from the Animal Experimental Ethics Committee of Chengdu Sport University (approval No. 2018024A).
    RESULTS AND CONCLUSION:  After 8 weeks of exercise, compared with the quiet control group, fasting blood glucose and HOMA-IRI in all T2DM rats increased significantly (P < 0.01). Compared with the quiet T2DM group, fasting blood glucose and insulin resistance index in the two T2DM exercise groups were significantly reduced (P < 0.01), while fasting blood glucose in the T2DM resistance exercise group was significantly lower than that of T2DM aerobic exercise group (P < 0.05). Compared with the quiet control group, the structure of the sciatic nerve in all T2DM rats showed typical demyelination changes, including disordered and loose myelin sheath arrangement, lamellar separation and shrinkage. Compared with the quiet T2DM group, the two T2DM groups had tighter myelin structure of the sciatic nerve and milder lamellar separation. Compared with the quiet control group, the MWT and TWL in the quiet T2DM group and T2DM resistance exercise group declined significantly (P < 0.01), whereas the TWL in the T2DM aerobic exercise group was significantly higher than that in the T2DM resistance exercise group (P < 0.05). Compared with the quiet control group, the expression of Grp78, Chop, and Caspase-12 in the quiet T2DM group was significantly increased (P < 0.01). Compared with the quiet T2DM group, the expression of Grp78, Chop, and Caspase-12 in the T2DM aerobic exercise group was significantly decreased (P < 0.01; P < 0.05). The expression of Grp78 and Caspase-12 in the T2DM resistance exercise group was significantly higher than that in the T2DM aerobic exercise group (P < 0.05). To conclude, endoplasmic reticulum stress in the peripheral nerve of diabetic rats results in demyelination of peripheral nerve and decrease of MWT and TWL in diabetic rats. Both aerobic exercise and resistance exercise can reduce the endoplasmic reticulum stress in the peripheral nerve of diabetic rats, and improve the structure and function of the sciatic nerve. Moreover, the effect of aerobic exercise on the endoplasmic reticulum stress is better than that of resistance exercise.

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    Effect of Capparis spinosa total alkaloid on proliferation and apoptosis of nucleus pulposus cells in an intervertebral disc degeneration rat model
    Liu Zhigang, Guo Qinggong, Chen Jingtao
    2021, 25 (11):  1699-1704.  doi: 10.3969/j.issn.2095-4344.3082
    Abstract ( 63 )   PDF (838KB) ( 12 )   Save
    BACKGROUND: Capparis spinosa total alkaloids (CSTA) have certain effects on cell growth and extracellular matrix synthesis. The aging and apoptosis of nucleus pulposus cells are one of the main pathologies of intervertebral disc degeneration. Therefore, it is assumed that CSTA may have certain effect on the degeneration of the intervertebral disc.
    OBJECTIVE: To study the effect of CSTA on intervertebral disc degeneration rat model and nucleus pulposus cells. 
    METHODS: Thirty-two Sprague-Dawley rats were randomly divided into sham, model, CSTA-H, and CSTA-L groups with eight in each group. Rat models of intervertebral disc degeneration were made in the model group, CSTA-L group and CSTA-H group. The CSTA-L and CSTA-H groups were given intragastric administration of CSTA 225 mg/kg/d and 450 mg/kg/d for 4 weeks respectively. Hematoxylin-eosin staining was used to observe the pathological changes of the intervertebral disc. Immunohistochemistry and western blot were used to detect the expression of type II collagen and aggrecan. Nucleus pulposus cells from the intervertebral disc of another two Sprague-Dawley rats were separated, cultured and divided into a control group, an oxygen-glucose deprivation (OGD) group and an administration group (OGD+CSTA 10 mg/L). After being cultured for 24 hours, the morphology of nucleus pulposus cells was observed, the cell proliferation ability was detected by cell counting kit-8, the cell apoptosis was detected by flow cytometry, and the expression of type II collagen and aggrecan were detected by western blot. 
    RESULTS AND CONCLUSION: (1) Compared with the sham group, the intervertebral disc tissue of the model group showed fiber ring fissures, aggregation and shrinking of nucleus pulposus cells, and different improvements were found in the CSTA-L and CSTA-H groups. (2) The expression levels of type II collagen and aggrecan in the model group were significantly lower than those in the sham, CSTA-L and CSTA-H groups (P < 0.05). (3) Compared with the control group, the cells in the OGD group showed irregular morphology and death status, whereas the cell morphology in the administration group was improved. (4) Compared with the control group, nucleus pulposus cells in the OGD group showed lower proliferation, higher apoptotic rate, and lower levels of type II collagen and aggrecan (P < 0.05). Compared with the OGD group, nucleus pulposus cells in the administration group showed faster proliferation, lower apoptotic rate, and higher levels of type II collagen and aggrecan. To conclude, CSTA can improve intervertebral disc degeneration by promoting proliferation and inhibiting apoptosis of nucleus pulposus cells as well as inhibiting the degradation of extracellular matrix.

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    Effect of cassia seed aqueous extract on blood pressure level in N-nitro-L-arginine-methyl ester induced hypertensive rats
    Ye Quanying, Chen Qisheng, Li Yanwen, Wang Ting, Chen Xiaoyan, Yue Yun
    2021, 25 (11):  1705-1711.  doi: 10.3969/j.issn.2095-4344.3105
    Abstract ( 85 )   PDF (1016KB) ( 33 )   Save
    BACKGROUND: The concept of modern diagnosis and treatment of hypertension is to protect target organs, improve clinical symptoms and minimize clinical events through antihypertensive treatment. Cassia seed has an antihypertensive effect that has been confirmed in animal experiment and clinical practice, but whether it can improve the vascular dysfunction, oxidative stress and end organ damage of hypertension remains to be further studied. 
    OBJECTIVE: To evaluate the antihypertensive effect of cassia seed water extract on hypertensive rats induced by N-nitro-L-arginine methyl ester. 
    METHODS: A rat model of hypertension was established by intragastric administration of N-nitro-L-arginine methyl ester to observe the changes of blood pressure and heart rate after the intervention of cassia seed aqueous extract (500 mg/kg per day), once a day, for 4 continuous weeks. Blood pressure and heart rate were measured once a week. At 24 hours after the final administration, serum levels of aspartate aminotransferase, alanine aminotransferase, urea and serum creatinine, and lipid mass spectrometry were detected by an automatic biochemical analyzer. Malondialdehyde, nitric oxide, reduced glutathione levels and angiotensin converting enzyme activity in rat liver and kidney were determined by corresponding assay kits. The expression levels of endothelial nitric oxide synthase and inducible nitric oxide synthase protein and gene in rat kidney tissues were examined by immunohistochemistry as well as real-time quantitative PCR. 
    RESULTS AND CONCLUSION: Compared with the model group, cassia seed aqueous extract treatment significantly reduced mean arterial pressure, diastolic pressure and systolic pressure (P < 0.05) in hypertensive rats, and improved liver and renal markers, lipid distribution and oxidative status. In addition, cassia seed aqueous extract significantly reduced the activity of angiotensin-converting enzyme in the liver and kidney of hypertensive rats (P < 0.05). The protein and mRNA expression levels of endothelial nitric oxide synthase in the kidney of rats treated with cassia seed aqueous extract were significantly higher than those of the model group. In conclusion, cassia seed water extract has shown considerable potential for antihypertension, and its anti-hypertension mechanism includes up-regulation of endothelial nitric oxide synthase expression, antioxidants and inhibition of angiotensin-converting enzymes.

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    Transgenic tomato vaccine expressing chimeric protein PAcA/CTB can immunize rats through gastrointestinal absorption
    Guan Weiwei, Gu Yu, Guan Xiaoyan, Wu Jiayuan, Bai Guohui, Tian Yuan, Liu Jianguo
    2021, 25 (11):  1712-1716.  doi: 10.3969/j.issn.2095-4344.3089
    Abstract ( 82 )   PDF (751KB) ( 69 )   Save
    BACKGROUND: The transgenic plant anti-caries vaccine has gradually demonstrated its unique advantages due to its simple production process, low price, good immunogenicity, safety and effectiveness, and has become a research focus in recent years.
    OBJECTIVE: To immunize the Sprague-Dawley rats with transgenic tomatoes carrying chimeric protein of PAcA/CTB by gavage and to observe the immunogenicity and immunoreactivity in the rats.
    METHODS: Male Sprague-Dawley rats were randomly divided into a positive control group, an experimental group and a negative control group, in which the rats were given deactivated Streptococcus mutans, transgenic tomatoes carrying PAcA/CTB, and normal tomatoes by gavage, respectively, at 1 week after establishment of dental caries models in animals. The intragastric administration was done once a week for 4 continuous weeks. Blood and saliva samples of rats were collected before immunization and 1, 2, 3, and 4 weeks after immunization to detect the levels of IgG and sIgA antibodies against Streptococcus mutans PAcA by ELISA. The upper and lower molars of the rats at the age of 57 days were scored for dental caries. The study protocol was approved by the Laboratory Animal Ethics Committee of Zunyi Medical University. 
    RESULTS AND CONCLUSION: Compared with the negative control group, positive control and experimental groups had a significant increase in the levels of anti-PAcA sIgA in saliva at 1, 2, 3, and 4 weeks after immunization (P < 0.01). Compared with the negative control group, the positive control group showed a significant increased level of anti-PAcA IgG in serum at 1-4 weeks after immunization (P < 0.05), while the experimental group had a significantly increased level of anti-PAcA IgG in serum at 2-4 weeks after immunization (P < 0.01). Tooth damage was minimized, only a few of caries reached the superficial dentin layer, and the number of caries was the least in the positive control group, followed by the experimental group and the negative control group in turn. To conclude, the transgenic tomatoes expressing PAcA/CTB has certain immunogenicity and immunoreactivity which can effectively immunize the rats by gavage and reduce the incidence of dental caries.

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    Oscillating field stimulation inhibits neuroinflammatory response and promotes functional recovery after spinal cord injury in rats
    Fang Chao, Sun Jian, Wei Laifu, Gao Fei, Qian Jun
    2021, 25 (11):  1717-1722.  doi: 10.3969/j.issn.2095-4344.3088
    Abstract ( 56 )   PDF (1299KB) ( 19 )   Save
    BACKGROUND: Oscillating field stimulation (OFS) is an emerging technology that has been used to treat spinal cord injury (SCI) with promising therapeutic effects. The application of electric current to the injured spinal cord is known to promote healing and tissue regeneration. However, the mechanisms by which OFS leads to SCI recovery are not yet fully understood.
    OBJECTIVE: To investigate whether OFS can promote the recovery of neurological function by reducing neuroinflammatory responses after SCI in rats.
    METHODS: Sixty-three female Sprague-Dawley rats were randomized into an experimental group, a SCI group and a sham operation group. Rats in the sham operation group only underwent laminectomy without damaging the spinal cord, and animal models of thoracic (T9-T10) SCI were established in the SCI and experimental groups. The OFS device was implanted into the injured site in the experimental group, followed by continuous electrical stimulation. Basso-Bettie-Bresnahan score was used to evaluate the recovery of motor function after SCI in rats. In addition, hematoxylin-eosin staining was used to test tissue repair and inflammatory cell recruitment. The number of astrocytes and microglia was examined by immunofluorescence staining. The study protocol was approved by the Animal Ethics Committee of Anhui Medical University, with an approval No. LLSC20190736.
    RESULTS AND CONCLUSION: The Basso-Bettie-Bresnahan score of the experimental group was significantly higher than that of the SCI group. Hematoxylin-eosin staining results showed that the number of inflammatory cells in the injured site was considerably reduced in the experimental group compared with the SCI group. The results of immunofluorescence detection showed that the number of glial fibrillary acidic protein positive cells and microglia in the experimental group was dramatically lower than that in the SCI group at different time after surgery. These findings indicate that OFS can improve the motor function of lower limbs, promote tissue repair, reduce inflammatory cell recruitment, and inhibit the activation of astrocytes and microglia after spinal cord injury in rats. OFS can promote the recovery of neurological function by inhibiting inflammatory response after SCI.  

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    Effects of moderate treadmill exercise on alpha-smooth muscle actin and type IV collagen in the liver of type 2 diabetic rats
    Hu Sheng, Yuan Haiyan, Hu Meng, Jin Shanhu
    2021, 25 (11):  1723-1727.  doi: 10.3969/j.issn.2095-4344.3074
    Abstract ( 48 )   PDF (758KB) ( 14 )   Save
    BACKGROUND: Diabetes-induced liver damage is easy to be ignored in the early stage. Exercise therapy can increase the sensitivity of insulin, which is an important means of prevention and treatment of diabetes.
    OBJECTIVE: To explore the effect of moderate intensity exercise intervention on liver injury during the occurrence of diabetes mellitus.
    METHODS: The experimental protocol was approved by the Laboratory Animal Care Ethics Committee of Wuhan Sports University. Thirty SPF Sprague-Dawley rats were divided into three groups: normal control group, diabetic control group and diabetic exercise intervention group. Normal control group was fed with normal diet, with no exercise. Diabetes control group was fed with high sugar and high fat diet for 8 weeks, followed by a small dose of streptozotocin (30 mg/kg) injected intraperitoneally, with o exercise. Diabetes exercise intervention group was fed and injected in the same way as diabetes control group, and at the same time carried out moderate intensity treadmill training. After 7 days of streptozotocin injection, hematoxylin-eosin and Masson staining were used to observe the cell morphology, the expression of α-smooth muscle actin was observed by immunohistochemistry, and orbital blood samples were collected to detect the concentration of serum type IV collagen using ELISA.
    RESULTS AND CONCLUSION: Compared with the normal control group, hematoxylin-eosin staining showed that the structure of liver lobule in the diabetic control group was disordered, a large number of fat vacuoles were seen, and the bile duct in the portal area was obviously proliferated; Masson staining that there were showed fat vacuoles, the structure of liver lobule was seriously damaged, and blue stained collagen fibers were seen in the portal area, and light blue stained collagen fibers were seen between liver cells. The above pathological changes were alleviated in the diabetic exercise intervention group, and the fatty degeneration of liver cells was obviously reduced. The expression of α-smooth muscle actin in the diabetic control group and diabetic exercise intervention group was significantly higher than that in the normal control group (P < 0.05), and that in the diabetic exercise intervention group was significantly lower than that in the diabetic control group (P < 0.05). The level of type IV collagen in the diabetic exercise intervention group was significantly lower than that in the diabetic control group (P < 0.05), to slow down the progress of fibrosis. To conclude, moderate intensity exercise has a good effect on streptozotocin induced liver fibrosis in diabetic rats.

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    Mechanism of hyperhomocysteinemia induced renal injury in Cbs+/- mice
    Wu Kai, Liu Kun, Xie Lin, Lu Guanjun, Ma Shengchao, Li Guizhong, Cao Jun, Jie Yuzhen, Jiang Yideng, Hao Yinju
    2021, 25 (11):  1728-1732.  doi: 10.3969/j.issn.2095-4344.3084
    Abstract ( 73 )   PDF (750KB) ( 32 )   Save
    BACKGROUND: In chronic kidney disease, there is often an increase in the level of homocysteine, which can lead to podocyte apoptosis, but the specific mechanism is not clear. 
    OBJECTIVE: To investigate the mechanism of hyperhomocysteinemia-induced renal injury in Cbs+/-  mice. 
    METHODS: Cbs+/+ mice (control group) and Cbs+/- mice (model group) with similar body weight were selected, with 10 mice in each group, and were fed with high methionine diet. After 8 weeks, the mice were killed, the serum was separated and the kidney tissue was obtained. The levels of serum homocysteine, urea nitrogen and creatinine were measured by automatic biochemical analyzer. The renal injury was observed by Periodic Acid-Schiff staining and transmission electron microscope. TUNEL staining was used to observe the apoptosis of glomeruli. The protein expression levels of Bax, Bcl-2 and caspase12 were detected by western blot. 
    RESULTS AND CONCLUSION: Compared with Cbs+/+ mice, the level of serum homocysteine, urea nitrogen and creatinine in Cbs+/-  mice were significantly increased (P < 0.01). The Periodic Acid-Schiff staining results showed that the glomerular basement membrane of Cbs+/+ mice was clear and the thickness was uniform, while the Cbs+/-  mouse glomerular basement membrane showed varying degrees of uneven thickness, widening of membrane area and thickening of matrix. Under the transmission electron microscope, the glomerular basement membrane of Cbs+/+ mice was clear and the foot process was regular, while the glomerular basement membrane of Cbs+/-  mice was locally thickened and the foot process was irregular fusion. TUNEL staining showed that the number of apoptotic cells in glomeruli of Cbs+/-  mice was significantly increased compared with Cbs+/+ mice; meanwhile, western blot detection showed that the protein levels of Bax/Bcl-2 and caspase12 were significantly increased (P < 0.05). To conclude, podocyte apoptosis plays an important role in hyperhomocysteinemia-induced renal injury in Cbs+/- mice.

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    Change of Ghrelin-GHSR pathway in 4-week intermittent hypoxic exposure improving obesity in mice
    Yang Luyao, Fu Pengyu, Tang Shuning, Zhu Rongxin, Gong Lijing
    2021, 25 (11):  1733-1739.  doi: 10.3969/j.issn.2095-4344.3098
    Abstract ( 99 )   PDF (923KB) ( 21 )   Save
    BACKGROUND: Weight loss under hypoxic exposure is related to a reduction in food intake. Decreased gastric Ghrelin levels can reduce food intake, but whether digestive nutritional absorption disorders and dyslipidemia caused by obesity can be regulated through Ghrelin-growth hormone secretagogue receptor (GHSR) pathway under intermittent hypoxia is unclear yet.
    OBJECTIVE: To explore the effect of intermittent hypoxia exposure on Ghrelin-GHSR pathway in gastric tissues of obese mice. 
    METHODS: Thirty C57BL/6J male mice were randomly divided into a control group (n=8) and a high fat diet group (n=22, fed with high fat diet). After 8 weeks high fat diet, obesity models were successfully established in 16 mice with high fat diet, and then were randomly divided into an obese control group (n=8) and an obese hypoxic group (n=8, 11.2% oxygen concentration, 8 hours per day, 6 times per week). The mice whose remaining body mass did not exceed the average body weight of the normal control group by 20% were discarded. After intervention, the serum biochemical indicators and Ghrelin level were tested; the Ghrelin and GHSR-1a mRNA expressions were tested by RT-PCR; gastric tissue morphology was observed by hematoxylin-eosin staining; the mean absorbance values of Ghrelin and Ghrelin O-acyltransferase (Goat) were measured by immunohistochemistry; the protein expressions of hypothalamic growth hormone secretagogue hormone receptor 1a (GHSR-1a), Ghrelin and Goat were detected by western blot. The study protocol was approved by the Sports Science Ethics Committee of Beijing Sport University, approval No. 2015021.
    RESULTS AND CONCLUSION: (1) The body mass of the obese control group was significantly higher than that of the control group and obese hypoxic group after 4-week hypoxic intervention. (2) The blood glucose, total cholesterol, and triacylglycerol levels were obviously higher in the obese control group than the control group. Compared with the obese control group, the blood glucose and total cholesterol levels were significantly lower in the obese hypoxic group, while the serum Ghrelin level was significantly higher in the obese hypoxic group. (3) The Ghrelin mRNA relative expression in gastric tissue of the obese control group was evidently lower than that of the obese hypoxic group, and the GHSR-1a mRNA expression in the hypothalamus was significantly lower in the obese control group than the control group. (4) The obese hypoxic group had a regular, dense and uniform distribution of gastric fundus gland main cells. (5) The mean absorbance values of Ghrelin and Goat as well as the protein contents of GHSR-1a, Ghrelin and Goat in the obese hypoxic group were evidently higher than those in the obese control group. These results indicate that 4-week hypoxic exposure can significantly increase the expression of Ghrelin and Goat in gastric tissue and GHSR-1a in the hypothalamus, decrease blood glucose level and body mass, and improve the adverse effects of obesity in obese mice.

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    Correlation of CY3A5 genetic polymorphism with concentration/dosage of tacrolimus and individualized administration of tacrolimus after kidney transplantation
    Liu Junchang, Gao Xiaolin, Jiang Taimao
    2021, 25 (11):  1740-1744.  doi: 10.3969/j.issn.2095-4344.3037
    Abstract ( 61 )   PDF (963KB) ( 36 )   Save
    BACKGROUND: Polymorphism of CYP3A5 gene can significantly affect the blood concentration of tacrolimus in the early period after kidney transplantation. Many studies in China are limited to the early 3 months after kidney transplantation with no concern on the long-term effect of tacrolimus in recipients.
    OBJECTIVE: To investigate the relationship between the polymorphism of CYP3A5 gene and tacrolimus concentration/dose (C0/D) in kidney transplant recipients, and to compare the differences among different genotypes, so as to provide an individualized drug regimen of tacrolimus after kidney transplantation. 
    METHODS: Sixty-five adult recipients who underwent kidney transplantation and postoperative administration of tacrolimus (FK506) + mycophenolate mofetil (MMF) + prednisone (Pred) immunosuppressive therapy were divided into three groups according to their CYP3A5 genotypes detected preoperatively: CYP3A5*1/*1, *1/*3, and *3/*3 groups.  The whole blood concentration of tacrolimus was monitored in all the recipients, and C0/D value was recorded in each group at different time points after surgery. The study protocol was in line with the ethical requirements of Air Force Hospital of Northern Theater Command. 
    RESULTS AND CONCLUSION: There were 6, 25 and 34 recipients of CYP3A5*1/*1, *1/*3 and *3/*3, respectively. The C0/D value of tacrolimus in the CYP3A5*1/*1 and *1/*3 groups was significantly lower than that in the CYP3A5*3/*3 group (P < 0.05). At 7 and 14 days after surgery, the C0/D value of tacrolimus in the CYP3A5*1/*1 group was lower than that in the CYP3A5*1/*3 group (P=0.028, P=0.034). In the CYP3A5*1/*1 group, the C0/D value of tacrolimus at 7 days after surgery was significantly lower than that at 6 months and 1 year after surgery (P=0.35, P=0.41). In the CYP3A5*1/*3 group, the C0/D value of tacrolimus at 7 days after surgery was significantly lower than that at 3 months, 6 months and 1 year after surgery (P=0.029, P=0.07, P < 0.01), and that at 14 days and 1 month after surgery was significantly lower than that at 6 months and 1 year after surgery (P=0.04, P=0.39). In the CYP3A5*3/*3 group, the C0/D value of tacrolimus at 7 days after surgery was significantly lower than that at 3 months, 6 months and 1 year after surgery (P=0.029, P=0.03), and that at 14 days after surgery was significantly lower than that at 6 months and 1 year after surgery (P=0.022). Overall findings indicate that the polymorphism of CYP3A5 gene has a significant effect on the C0/D value of tacrolimus in kidney transplant recipients, which can be maintained for a long-term stable period after transplantation. For CYP3A5*1/*1 and *1/*3 recipients, the metabolism of tacrolimus is faster in the early stage, and the dosage of tacrolimus should be increased to maintain the target blood concentration, whereas for CYP3A5*1/*3 recipients, the dosage of tacrolimus may be moderately less than the former and the drug reduction rate should be slowed down in the later stage. CYP3A5*3/*3 recipients have a slow metabolism of tacrolimus, and should be given a small dose in the early stage, and the reduction rate should be accelerated in the later stage.

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    Knowledge network mapping of literature regarding platelet-rich plasma in recent 5 years: a visual analysis
    Tian Yanping, Li Juan, Liu Xiaobo, Zhang Huiling, Shi Lihong, Jin Rongjiang
    2021, 25 (11):  1745-1752.  doi: 10.3969/j.issn.2095-4344.3090
    Abstract ( 62 )   PDF (1239KB) ( 22 )   Save
    BACKGROUND: Platelet-rich plasma has been widely used in orthopedics, plastic surgery, maxillofacial surgery, sports medicine, and intractable wounds due to its advantages of simple preparation, autologous source, low cost, and good safety.
    OBJECTIVE: To analyze the existing research structure of platelet-rich plasma in recent 5 years worldwide by using CiteSpace software, and to analyze the research hotspots and the evolution trend of platelet-rich plasma by using the visual network map generated by the software, in order to explore the academic frontiers and provide the choice of research direction. 
    METHODS: The Web of Science Core Collection and CNKI database were searched for platelet-rich plasma related literature records published between 2015 and 2020. We visually analyzed the enrolled studies in terms of countries/regions, institutions, authors, journals, and keywords based on the knowledge network maps generated by the CiteSpace software. 
    RESULTS AND CONCLUSION: The average annual number of documents issued in China from 2015 to 2019 was 184.2; the high-yield institutions issued in cooperation were represented by the Department of Orthopaedics of the Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University; the high-yield authors issued in cooperation were represented by Shan Guiqiu; the keywords of hotspots were growth factors, bone marrow stem cells, osteoarthritis, etc. Internationally, the average annual number of documents issued in 2015-2019 was about 928.2; the countries with high yield of cooperative documents issued were the United States, China and Italy; the institutions with high yield of cooperative documents issued were New York Hospital for Special Surgery, University of Pittsburgh and Rush University; the author with high yield of cooperative documents issued was Anitua E; the keywords for hotspots were growth factors, stem cells and repair; and the influential journal was American Journal of Sports Medicine. According to the results of CiteSpace literature analysis, platelet-rich plasma has developed rapidly in the fields of orthopedics, sports medicine and tissue engineering, both at home and abroad. 

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    Thymosin alpha1 protects against liver injury in rats with zymosan-induced multiple organ failure
    Ma Minghe, Niu Yi
    2021, 25 (11):  1753-1758.  doi: 10.3969/j.issn.2095-4344.3076
    Abstract ( 42 )   PDF (1151KB) ( 25 )   Save
    BACKGROUND: Liver injury in a multiple organ failure model causes great troubles to clinicians’ medication. Thymosin α1 is used for treating chronic hepatitis and it has obvious protective effects against liver injury.
    OBJECTIVE: To investigate the protective mechanism of thymosin α1 on liver injury in a rat model of multiple organ failure, based on adiponectin (ADPN)/protein kinase B (Akt)/nuclear factor κB (NF-κB) signaling pathway. 
    METHODS: Male SPF Sprague-Dawley rats were randomly divided into four groups: normal group, model group, experimental group, and control group. Rats in the model group, experimental group, and control group were given intraperitoneal injection of 500 mg/kg zymosan (50 g/L) to construct the rat multiple organ failure model. Normal rats were injected intraperitoneally with equal doses of normal saline. Thirty minutes after the injection, the rats in the experimental group and the control group were injected intraperitoneally with 2 mL of thymosin α1 and ganlixin with the dose of 0.5 mg/kg daily, respectively. The normal group and the model group were injected intraperitoneally with the same dose of normal saline. After 7 days of continuous administration, liver function parameters were tested;  histopathological changes of rat liver tissues and cell apoptosis were detected using hematoxylin-eosin staining and TUNEL staining; immunohistochemistry and western blot were used to detect the expression of interleukin-10, tumor necrosis factor α (TNF-α), adiponectin (ADPN), adiponectin recepror 2, AdipoR2, p-AKT and NF-κB. 
    RESULTS AND CONCLUSION: Compared with the normal group, the levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin in the serum, the pathological scores of liver injury, the cell apoptotic rate, and the expression levek of TNF-α and NF-κB were significantly increased in the model group, while the serum levels of total protein, interleukin-10, ADPN, AdipoR2 and p-AKT were significantly reduced in the model group (all P < 0.05). Compared with the model group, the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, the pathological scores of liver injury, and cell apoptotic rate in the experimental group and control group were significantly reduced, and the serum levels of total protein, interleukin-10, ADPN, AdipoR2 and p-AKT were significantly increased (all P < 0.05). To conclude, thymosin α1 has a protective effect on the liver of rats with multiple organ failure induced by zymosan. The mechanism is related to the ADPN/Akt/NF-κB signaling pathway. ADPN/Akt is activated and the activation of NF-κB is inhibited, then reducing the inflammatory response. 

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    The role of endoplasmic reticulum stress in the pathogenesis of steroid-induced avascular necrosis of the femoral head 
    Zhang Xujian, Zhao Zhenqun, Liu Wanlin
    2021, 25 (11):  1759-1765.  doi: 10.3969/j.issn.2095-4344.3104
    Abstract ( 45 )   PDF (556KB) ( 10 )   Save
    BACKGROUND: The pathogenesis of steroid-induced avascular necrosis of the femoral head is not clear, which may be related to endoplasmic reticulum stress. Controlling endoplasmic reticulum stress signal pathway may regulate autophagy and apoptosis, which has a certain preventive and therapeutic effect on the disease.
    OBJECTIVE: To explore the relationship between endoplasmic reticulum stress and hormone-induced autophagy and apoptosis, and to summarize the research progress of endoplasmic reticulum stress in the pathogenesis of steroid-induced avascular necrosis of the femoral head.
    METHODS: Relevant articles published from 2000 to 2020 were retrieved from PubMed, Web of Science, CNKI, WanFang, and VIP databases. The keywords were “endoplasmic reticulum stress, femur head necrosis, unfolded protein response, glucocorticoid, autophagy, apoptosis, ischaemia” in English and Chinese, respectively. The old and duplicate articles were excluded, and 81 articles were included for analysis and discussion.
    RESULTS AND CONCLUSION: Unfolded protein response can induce autophagy and apoptosis in cells to alleviate the three downstream signal pathways caused by endoplasmic reticulum stress. Endoplasmic reticulum stress is closely related to hormone-induced autophagy and apoptosis and femoral head ischemia. Endoplasmic reticulum stress may be a pathological link in the process of steroid-induced avascular necrosis of the femoral head. Hormone induces autophagy and apoptosis of cells by activating endoplasmic reticulum during ischemia and hypoxia and finally leads to osteonecrosis of the femoral head.

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    Posterior cruciate ligament injury: diagnosis, treatments and rehabilitation strategies
    Xu Baoyun, Fu Weili
    2021, 25 (11):  1766-1772.  doi: 10.3969/j.issn.2095-4344.3109
    Abstract ( 98 )   PDF (759KB) ( 18 )   Save
    BACKGROUND: With the increasing cases of posterior cruciate ligament injury, there are endless protocols for diagnosing and treating posterior cruciate ligament injury. However, there are few reviews of the integrity of posterior cruciate ligament. 
    OBJECTIVE: To complete a comprehensive review of posterior cruciate ligament injury in terms of anatomy, clinical manifestations, auxiliary examination, treatment methods and rehabilitation strategies.
    METHODS: The PubMed, Ovid, CKNI, and WanFang databases were retrieved using the key words of “posterior cruciate ligament, PCL, anatom*, diagnos*, treatments, surger*, rehabilitation.” A total of 223 articles were searched. After removal of repetitive and ineligible literature, 65 articles were included for review.
    RESULTS AND CONCLUSION: Missed diagnosis and misdiagnosis of posterior cruciate ligament injury certainly exist. Posterior cruciate ligament injuries are mostly caused by traffic injuries and sports injuries, and are accompanied by other structural injuries to a certain degree. A full understanding of patient’s medical history, accurate physical examination methods, and sophisticated auxiliary examinations can help to correctly identify injuries to the posterior cruciate ligament and the surrounding structures, so as to formulate a reasonable diagnosis and treatment protocol. Research suggests that patients with posterior cruciate ligament injury should be subjected to reconstruction of the posterior cruciate ligament as soon as possible to reduce the risk of further degeneration. At present, there are still large disagreements on the diagnosis, treatment methods, and rehabilitation strategies of posterior cruciate ligament injury. So, a large number of rigorous randomized controlled trials are urgently needed to select the most suitable diagnosis and treatment methods.

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    Bioinformatics analysis of gene expression profile and key pathways related to fatty infiltration after rotator cuff injury
    Liu Peidong, Feng Jiangfeng, Xu Wenjie, Xu Xiaodong, Yang Ziquan
    2021, 25 (11):  1773-1778.  doi: 10.3969/j.issn.2095-4344.3083
    Abstract ( 77 )   PDF (2214KB) ( 13 )   Save
    BACKGROUND: Fat infiltration is a key factor in the failure of rotator cuff repair. However, the pathological mechanism of fatty infiltration after rotator cuff injury is unclear. 
    OBJECTIVE: To explore the differences in the expression of key genes after rotator cuff injury, to determine their functions and mechanism pathways, and to provide a theoretical basis for the pathological mechanism of fatty infiltration after rotator cuff injury. 
    METHODS: GSE93661 was obtained through GEO database to screen differentially expressed genes. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to analyze the underlying mechanism of fatty infiltration. The protein-protein interaction network was constructed to obtain the pivot genes and analyze the potential pathogenic targets. 
    RESULTS AND CONCLUSION: A total of 471 differentially expressed genes were identified. GO and KEGG analysis showed that neuroactive ligand-receptor interactions and cell adhesion molecular pathways were potential mechanisms of fat infiltration in rotator cuff tears. Leukotriene B4 receptor, as a pivot gene in the protein-protein interaction network, may be a key target for fat infiltration in rotator cuff tears. We have discovered potential key genes and pathways in the pathological development of fatty infiltration, providing a reference direction for future treatment.

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    MicroRNA-214 is involved in the regulation of bone metabolism
    Li Dongdong, Liao Hongbing
    2021, 25 (11):  1779-1784.  doi: 10.3969/j.issn.2095-4344.3080
    Abstract ( 78 )   PDF (1123KB) ( 24 )   Save
    BACKGROUND: MicroRNAs (miR) is an important endogenous non-coding small RNA that regulates the expression of genes by regulating the translation of mRNA. In recent years, it has been found that miR-214 plays an important role in related bone metabolic signaling pathways, which can regulate bone resorption and bone formation by targeting related genes. 
    OBJECTIVE: To review the new progress in the regulatory mechanism and possible application of miR-214 in bone metabolism.
    METHODS: The first author searched PubMed, Web of Science, and Medline with the keywords of “miRNA, miR-214, osteogenesis, osteoblast,” “miR-214, bone remodeling,” and “miR-214, osteoclast, tissue engineering” respectively for relevant literature published from 2005 to 2020. A total of 761 articles were preliminarily searched, and 63 articles that were related to the research purpose were selected and analyzed.
    RESULTS AND CONCLUSION: MiR-214 can promote osteoclast differentiation by targeting phosphatase-tensin homolog and tumor necrosis factor receptor associated factor 3, and inhibit osteoblast differentiation by targeting transforming growth factor β activated kinase 1 binding protein 2, cadherin protein β1, Osterix, activating transcription factor 4, α1 type IV collagen, baculovirus IAP repeat containing 7, fibroblast growth factor receptor 1, TAFA chemokine-like family member 5, and bone morphogenetic protein 2. Many studies have proved that silencing or overexpression of miR-214 can regulate bone metabolism. It is also found that miR-214 in serum or extracellular vesicles may be a marker for the diagnosis and prognosis of some diseases. It is the focus of its application research to combine miR-214 antagonists with bioscaffold materials to form a stable, efficient and safe sustained release system. Therefore, miR-214 may have great potential in the treatment of bone metabolic diseases in the future.

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    Extracellular matrix and tissue engineering regeneration and repair
    Luo Yaxin, Bi Haoran, Chen Xiaoxu, Yang Kun
    2021, 25 (11):  1785-1790.  doi: 10.3969/j.issn.2095-4344.3100
    Abstract ( 146 )   PDF (669KB) ( 27 )   Save
    BACKGROUND: Extracellular matrix can provide a living environment for different types of cells in the body and affect their biological activities, playing an important role in maintaining cell homeostasis and tissue development.
    OBJECTIVE: To review the research progress of extracellular matrix and its application in tissue engineering regeneration.
    METHODS: Databases of PubMed and CNKI were retrieved with the key words of “extracellular matrix, tissue engineering, regeneration” in English and Chinese search from 2010 to 2020, respectively.  
    RESULTS AND CONCLUSION: The extracellular matrix contains a variety of components, and its extraction methods are diverse. It has many functions, such as guiding cells, transmitting signals, diagnosing and treating diseases, regenerating and repairing. As a precise and orderly network structure, extracellular matrix has unique advantages in tissue regeneration and repair. Extracellular matrix provides a microenvironment closest to the growth of cells in vivo, and it is rich in various active molecules, which can provide the basis for cell activity, and has a good application prospect. The forms of extracellular matrix existing in tissue regeneration are varied, and it has good biodegradability and low immunogenicity, which can also improve the inflammatory response to a certain extent, can support tissue regeneration and guide tissue reconstruction effectively. With the continuous in-depth study to extracellular matrix, its application in tissue engineering will be better developed, and it will also bring emerging technologies and treatment methods for regenerative medicine and tissue engineering in the future. However, as for the problems faced by extracellular matrix in tissue repair applications, further research and discussion are needed.

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    Transcriptome sequencing technology and its application in oral diseases, dental implants and regeneration
    Long Qian, Guan Xiaoyan, Wang Qian, Hu Huan, Liu Jianguo
    2021, 25 (11):  1791-1798.  doi: 10.3969/j.issn.2095-4344.3077
    Abstract ( 62 )   PDF (781KB) ( 33 )   Save
    BACKGROUND:  Since the application of transcriptome sequencing technology has made remarkable achievements in the research of melanoma and breast cancer, transcriptome sequencing technology has become a hot research method in scientific research and widely used in the field of stomatology.
    OBJECTIVE: To review the development of transcriptome sequencing technology and its application in various disciplines of stomatology through searching, screening and reading literature.
    METHODS: A search of CNKI, CBM and PubMed was performed for relevant literature published from 2015 through 2020. The search terms were “transcriptome, sequencing technology, RNA-seq, microarray, oral cancers, OSCC, periodontal, caries, pulp disease, tooth development, DPSCs, PDLSCs, orthodontics, implant” in English and Chinese, respectively. According to the inclusion and exclusion criteria, 72 literatures were included to review the development of transcriptome sequencing technology and its application in the field of stomatology.
    RESULTS AND CONCLUSION: In the development of transcriptome sequencing technology, RNA-seq technology has the greatest advantage in scientific research, because of its high accuracy, high throughput and low price. In the field of stomatology, this technology has been used in the research of oral squamous cell carcinoma, periodontitis, dental pulp disease, tooth regeneration, orthodontics and dental implantation, and has achieved some achievements. However, the current research on the same kind of disease does not reflect the relationship between the various studies, and the research content is limited. It is believed that more discoveries can be yielded in stomatology by exploring the relationship between different studies and expanding the research content.

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    Application of functional near-infrared spectroscopy in the study of depression
    Wang Liqun, Li Yuxi, Jin Rongjiang, Wang Wenchun, Pang Richao, Zhang Anren
    2021, 25 (11):  1799-1804.  doi: 10.3969/j.issn.2095-4344.3078
    Abstract ( 108 )   PDF (699KB) ( 20 )   Save
    BACKGROUND: The current diagnosis and study of depression lacks objective biological indicators. As an emerging non-invasive brain functional imaging technology, functional near infrared spectroscopy (fNIRS) may be a potential biomarker for the identification and diagnosis of depression.
    OBJECTIVE: To review the application of fNIRS technique in depression research and provide guidance for clinical research.
    METHODS: The keywords were “functional near-infrared spectroscopy, oxyhemoglobins, prefrontal cortex, depressive disorder.” The first author searched CNKI, WanFang, VIP, PuMed and Web of Science database from 1996 to 2020 in Chinese and English. Finally, 50 literatures were selected for review.
    RESULTS AND CONCLUSION: fNIRS signal may be a potential biomarker for depression. fNIRS technology is mainly used to study the cognitive dysfunction of depression based on the frontal lobe hemodynamic changes, which can be used for the differential diagnosis of depression. However, the current research on fNIRS is still in the exploratory stage, and there are still some defects in the existing research, which need to be further optimized and deepened in the future research. It can increase the study sample size and monitoring indicators, and integrate fNIRS with other technologies to improve the temporal and spatial resolution, so as to study the neural mechanism of depression systematically, comprehensively and accurately.

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