Objective To identify the availability of video-assisted thoracoscopic thymectomy. Methods Retrospectively analyzed 68 patients with myasthenia gravis who underwent thymectomy including 34 cases of video-assisted thoracoscopic thymectomy and 34 cases of transsternal thymectomy,and the patients were followed up for 16 years. Results Patients of the VATS group were given video-assisted thora-coscopic thymectomy. The mean operative time was (90. 1 ± 15. 0) min,mean blood loss was (45. 0 ± 5. 5) mL,mean chest tube drainage time was (2. 5 ± 1. 2) days and mean postoperative hospital stay was (7. 0 ± 1. 2) days in VATS group,and there was no serious complica-tions and surgical death. The mean operative time was (98. 0 ± 12. 5) min,mean blood loss was (118. 5 ± 17. 5) mL,mean chest tube drain-age time was (4. 5 ± 1. 3) days and mean postoperative hospital stay was (11. 0 ± 2. 5) days in transsternal thymectomy group. 3 patients de-veloped MG crisis. There was no significant difference in mid-and long-term effects between the two groups(P>0. 05). ConclusionVideo-as-sisted thoracoscopic thymectomy for MG is safe and feasible with the advantage of less invasion,less surgical trauma,lower rate of complica-tion,and good curative effect compared with transsternal thymectomy.

Objective To investigate the xenotransplantation model of fetal pig skin precursor tissue and its development after transplantaion. Methods Porcine skin precursor tissue was obtained from the embryo of gestation day 56 (E56), and made into microskin or stamp skin graft. The microskin was transplanted to the dorsal wound in BALB/c nude mice, then covered with human corpse skin. The stamp skin graft was imbedded subcutaneously into the back of nude mice, and microskin was injected subcutaneously into the auricles of nude mice. Their growth and development were observed and they were examined by HE staining at 6th and 12th week after transplantation respectively. Two-sample t test was used to analyze the size of newly grown skin tissue. Results Porcine skin precursor tissue graft in three models above survived and continued growth after transplantation, and growth ability of the dorsal wound transplantation model was significantly stronger than that of the auricle model. Epidermis and hypodermis were detected in newly grown skin tissues. Hair follicles, a few of sebaceous glands, but no sweat glands were observed in auricle model, while many sebaceous glands and sweat glands were observed in the dorsal wound model. Conclusion Transplantation of microskin to dorsal wound is the optimal model of investigating the xenotransplantation of fetal pig skin precursor tissue and its development after transplantion.

Objective Non-invasive detection is the focus of intense research in diagnosis of acute renal allograft rejection currently. Urine protein is considered the cue to reflect the pathological changes in kidney disease. In this study, we explored the urine markers for early acute renal allograft rejection. Methods The urine protein of two patients with acute renal allograft rejection were examined by 2D gel electrophoresis and bioinformatics. We adopted pH 4-7 ready strip IPG and stained the gel with Sypro-Ruby. The digitized 2D maps of urine protein were quantitatively analyzed using 2D-analysis software packages. By analyzing the differential expressions of proteome between different time points (1, 2, 3 days before acute rejection and 7, 14, 21 days after acute rejection), 30 protein spots were selected and analyzed by MALDI-TOF-MS/MS. Results We obtained 2D gel electrophoresis maps of urine protein of the patients with acute renal allograft rejection, which are of good reproducibility and resolution. Sixteen protein spots were identified, resulting in thirteen corresponding proteins. Out of these proteins, we screened three proteins (alpha-1-antichymotrypsin, tumor rejection antigen gp96, Zn-Alpha-2-Glycoprotein) closely related to acute rejection. Conclusion The urine protein spots on 2D gel electrophoresis maps for the patients with acute renal allograft rejection were of obvious difference when detected at different time points of acute rejection. Alpha-1-antichymotrypsin, tumor rejection antigen gp96 and Zn-Alpha-2-Glycoprotein might be the candidate protein markers to diagnose acute renal allograft rejection after renal transplantation.

Objective:To investigate and compare the effects of water extracts of Whitmania pigra Whitman and Hirudinaria ma-nillensis Lesson on the angiogenesis of Tg (kdrl:mCherry) zebrafish. Methods:The zebrafish embryos 6-8 hours after fertilization (6-8hpf) were transferred to the culture medium containing Whitmania pigra Whitman or Hirudinaria manillensis Lesson extract at different concentrations, and the culture medium containing the drugs was replaced every 24 h. And then, at 72 hpf, the larvalmorphology and intersegmental vessels were observed under a microscope. The hatchability of 48-and 72-hpf embryos, and the number of intersegmen-tal vessels and the heart rate of 72-hpf juveniles were measured. Results:Compared with the control group, when the concentration of Whitmania pigra Whitmanis was higher than 30μg· ml-1 , and the concentration of Hirudinaria manillensis Lesson was higher than 20μg· ml-1, the number of intersegmental vessels was significantly reduced (P<0. 01). Compared with the control group, at 48 hpf, when the concentration of the drug groups was higher than 40 μg· ml-1 , the hatchability of the two groups significantly decreased ( P<0. 01);at 72 hpf, the hatchability of Whitmania pigra Whitman decreased significantly at the concentration of 100 μg·ml-1 (P<0.01), while the hatchability of Hirudinaria manillensis Lesson decreased significantly at the concentration of 80 μg· ml-1(P <0. 01). There was no obvious yolk sac edema, pericardial edema and spine curvature in the two groups. The heart rate decreased sig-nificantly (P<0. 01), while was still within the normal range. Conclusion:Both Whitmania pigra Whitman and Hirudinaria manillen-sis Lesson have notable anti-angiogenic activity, and the anti-angiogenesis activity of Hirudinaria manillensis Lesson is stronger. They both have effects on the development of zebrafish embryos, while the toxicity is not obvious.

Objective To investigate the effect and molecular mechanism of the combination of mechanical strain stimulation and icariin (ICA) on inhibiting the differentiation of osteoclasts induced by fatigue load stimulation. Methods The mouse mononuclear macrophage cell line RAW264.7 was cultured in vitro, and the blank control group was α-MEM complete medium. In the fatigue load group, RAW264.7 cells were treated with 5000 μεmechanical stretch strain, and then cultured in an osteoclast culture medium that was an α-MEM complete medium containing 40 ng/ml macrophage colony-stimulating factor and 40 ng/ml osteoclast differentiation factor. In the mechanical stimulation + ICA group, RAW264.7 cells were treated as the same procedure in the fatigue load group, and then cultured in an α-MEM complete medium containing 1 ×10 -5 mol/L ICA simultaneously with a 1000 μεtensile strain on the substrate. The activity of tartrate-resistant acid phosphatase (TRAP) was detected using a TRAP assay kit. The mRNA expression of the osteoclast marker genes, i.e. TRAP, cathepsin K(CTSK) and matrix metalloproteinase 9 (MMP-9) was detected by real-time RT-PCR. The nuclear translocation of nuclear factor kappa B (NF-κB) was analyzed by Western Blot. Results Compared with the fatigue load group, the combination of mechanical stimulation (1000 με substrate stretching) and ICA (1×10-5 mol/L) could significantly inhibit the activity of TRAP in osteoclasts (P<0.01) and reduce osteoclastosis. Moreover, that combination not only could down-regulate the mRNA expression of TRAP, CTSK and MMP-9 and the differences were statistically significant (all P<0.01), but also could inhibit the formation of osteoclasts by inhibiting the phosphorylation of P65, P50 and IκB-α in NF-κB signaling pathway. Conclusions The coupling of mechanical stimulation and ICA can effectively inhibit the osteoclast differentiation and the bone resorption induced by fatigue load, and the mechanism may involve regulating NF-κB signaling pathway.

Intestinal mucosal barrier is the first line of defense against exogenous substances. A complete mechanical barrier can separate the body from the contents of the intestines and prevent the translocation of bacteria and microorganisms in the intestines, playing an important role in maintaining the homeostasis of the human body. If the intestinal mechanical barrier function is damaged, it will cause the intestinal endotoxin and bacterial translocation, leading to systemic inflammatory response syndrome and multiple organ dysfunction diseases. The barrier function of intestinal mucosa is a key link in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Therefore, the maintenance of intestinal mucosal barrier function is of great value for the diagnosis and treatment of NAFLD. In this paper, the research progress in intestinal mechanical barrier function of NAFLD was reviewed, and the main factors maintaining intestinal permeability of NAFLD were introduced, such as apoptosis, autophagy, apical junctional complex, cytoskeleton, main signal pathway, to provide reference for the diagnosis and treatment of NAFLD.

Objective:To explore the clinical features, diagnosis and prognosis of renal transplant recipients with COVID-19.Methods:The clinical data were retrospectively analyzed for 2 kidney transplant recipients with COVID-19. Based upon clinical manifestations, blood routine, inflammatory factors, cell immunity, chest computed tomography(CT)and therapeutic efficacies, the diagnosis and treatment of COVID-19 in kidney transplant recipients(Interim Edition V)were compared to that of ordinary COVID-19 patients. Both recipients had an onset of low/moderate fever. There was no initial symptom of cough or fatigue. Blood routine indicated a normal count of leukocytes, a marked lymphocytopenia, elevated C-reactive protein(CRP)and slightly higher procalcitonin(PCT). Cellular immunity was extremely low and chest CT showed multiple patchy ground-glass opacities in both lungs.Results:After 1 week of onset, both patients had a marked disease progression. The pathogenesis and imaging changes were highly similar to those reported for ordinary COVID-19 patients. For preventing secondary infections, both received symptomatic supportive measures of antiviral agents, withdrawing immunosuppressants, tapering of hormone maintenance dose, intravenous drip of gamma globulin and respiratory supports. Currently the conditions of both patients obviously improved and renal function was stable. One case recovered and was discharged.Conclusions:The clinical manifestations of COVID-19 in renal transplant recipients are generally consistent with that of ordinary COVID-19 patients. Although there is no established treatment for COVID-19, withdrawing immunosuppressants, maintaining small and medium doses of hormones, actively restoring immunity and providing respiratory supports in a timely manner are effective.

Objective:To investigate the possible mechanism of rubbing abdomen to regulate intestinal homeostasis in irritable bowel syndrome with constipation (IBS-C) mouse models.Methods:IBS-C mouse models of intestinal immune dysfunction were established using trinitrobenzene sulfonic acid (TNBS)-induced C57BL/J6 male mice. Thirty C57BL/J6 mice were randomly divided into three groups: the control group, the model group, and the mogul group. After 7 days of modeling, mice in the mogul group were given a mogul mechanical stimulation intervention once per day for 2 weeks, while mice in the control and model groups were not given any intervention. Changes in serum levels of interleukin-6 (IL-6) and IL-17A were detected by enzyme-linked immune sorbent assay (ELISA) and immunohistochemistry. The gene expression and protein levels of IL-17A and IL-23 were detected by quantitative real-time fluorescence PCR (qRT-PCR) and Western Blot, respectively. The morphological changes were observed by HE staining. The CD44 and CD62L expression changes were observed by immunofluorescence staining.Results:Compared with the model group, the levels of IL-6 and IL-17A in the serum of mice in the mogul group were decreased, and the expression of IL-6 and IL-7A in the tissues was down-regulated (all P<0.001). In addition, the gene expression and protein expression levels of IL-17A and IL-13 in the tissues of mice in the mogul group were decreased (all P<0.001). HE staining results showed that the mogul mechanical stimulation intervention could repair colonic tissues and reduce the inflammatory response. Immunofluorescence staining results showed that mogul mechanical stimulation intervention could downregulate the expression of CD44 but had no modulating effect on the expression of CD62L. Conclusions:Rubbing abdomen can improve intestinal homeostasis in IBS-C model mice by regulating changes in Th17 cell function.

Objective To explore the effect of peripheral blood genomic DNA methylation on the relationship between methyl donor status and risk of breast cancer.Methods A case-control study was conducted.Each three hundred breast cancer cases and controls were consecutively recruited.Food frequency questionnaire was used to collect dietary information.Amounts on folate,methionine,choline and betaine intake were calculated.Blood samples were collected for DNA extraction.Peripheral blood genomic DNA methylation was measured by using the Methyl FlashTM Methylated DNA Quantification Kit.Pathway analysis was used to examine the effect of genomic DNA methylation on the relations between methyl donor status and risk of breast cancer.Results The genome DNA methylation rates were 0.46％ ± 0.25％ and 0.53％ ± 0.34％,respectively on both cases and controls,with differences statistically significant (P＜0.01).Results from the pathway analysis,results showed that methionine consumption was related to genomic DNA methylation (β=0.065,P＜ 0.05) while genomic DNA methylation was related to the risk of breast cancerk (β =-0.027,P＜ 0.05),respectively.Conclusions The level of peripheral blood genomic DNA methylation in breast cancer cases was significantly lower than that in the controls.Genomic DNA methylation seemed to have played a mediated role between methionine and the risk of breast cancer.

Objective:To observe the structural and functional changes of each nucleus group in generalized anxiety disorder(GAD) rat models before and after the massage therapy, and to explore the central regulating mechanism of the massage therapy in GAD.Methods:60 Wistar male rats were randomly divided into blank group, model group and abdominal rubbing group, 20 rats for each group. GAD rat models were established by chronic emotional stress, and the models were evaluated by the elevated cross maze experiment. In the abdominal rubbing group, the GAD rats were treated with abdominal rubbing once a day for 10 min for 14 days. In the control group, the GAD rats were only bound to the experimental platform once a day for 10 min for 14 days. Horseradish peroxidase (HRP) retrograde tracking marker method was used to detect the changes of the structure of each nuclear cluster in the affective loop of the GAD rats in each group. The liquid-mass spectrometry quantitative analysis was used to determine the relative concentrations of N-acetyl aspartate (NAA), glutamic acid, choline and creatine in nuclear cluster of the GAD rats in each group.Results:Compared with the blank group, the HRP labeled cells in the rats of model group were less expressed and the activity of the emotional circuit in the brain was weak. The NAA/creatine ratio in the left hippocampal tissue of the rats in the model group was significantly increased than that of the blank group(0.94±0.08 vs 0.79±0.10, P<0.05), the glutamic acid/creatine ratio in the right hippocampal tissue was significantly decreased (0.95±0.10 vs 1.12±0.13, P<0.01), and the NAA/creatine andglutamic and acid/creatineratios in the left cortical tissue were all significantly decreased (1.04±0.05 vs 1.41±0.23, and 1.21±0.04 vs 1.57±0.11, all P<0.01). Compared with the model group, the hippocampal tissues of the GAD rats in the abdominal rubbing group had a large number of HRP aggregation and spread to the surrounding tissues, the NAA/creatine and glutamic acid/creatine ratios in the left hippocampal tissue of the abdominal rubbing group were significantly decreased (0.74±0.21 vs 0.94±0.09 and 0.92±0.20 vs 1.21±0.12, all P<0.01). The differences between the model group and the abdominal rubbing group in the ratio of glutamic acid/creatine in the right hippocampal tissue (1.01±0.23 vs 0.95±0.10), the ratio of NAA/creatine in the left cortical tissue (1.12±0.09 vs 1.04±0.05), and the ratio of glutamic acid/creatine in the left cortical tissue (1.22±0.12 vs 1.21±0.04) were not statistically significant ( P>0.05). Conclusions:Abdominal rubbing can effectively enhance the activity of affective loop and improve the relative concentration of neuron metabolites in hippocampal and cortical tissues in the affective loop, suggesting that the mechanism of GAD treatment by abdominal rubbing may be closely related to the structure and function of regulating affective loop.