Background and purpose:Rabs are members of Ras-related small GTPase superfamily. Rab27A is a unique member in the Rab family and has specific implications in human genetic diseases. We studied the potential role of Rab27A in proliferation, distribution of cell cycle, apoptosis and invasion of breast cancer cells and its mechanism(s). Methods:The eukaryotic expression vector containing Rab27A open reading frame (ORF) pcDNA3.1(+) - Rab27A was constructed and transfected into MDA-MB-231 breast cancer cells. Then we detected the changes in terms of cell growth, cell cycle distribution, apoptosis and in vitro invasion capability before and after transfection. We also applied RT-PCR to investigate the molecular basis.Results:① The expression of Rab27A was increased as invasive and metastatic ability increased in four human breast cancer cell lines. ② Overexpression of Rab27A can promote breast cancer cells to grow faster, increase the proportion of S phase cells, avoid apoptosis and invade in vitro. ③ Rab27A transfectants constitutively enhanced the expression of Cyclin D1, MMP-7 and MMP-9 in MDA-MB-231 cell lines, on the contrary, that of p16 were down-regulated constitutively. Reduced Rab27A expression by RNAi down-regulated the expression of Cyclin D1, MMP-7 and MMP-9, and up-regulated p16 expression.Conclusions:Rab27A can stimulate breast cancer cells to proliferate, increase the proportion of cells in S phase,avoid apoptosis and invade in vitro by regulating the expression of Cyclin D1, MMP-7, MMP-9 and p16.

Objective To determine the clinical characteristics in elderly patients with Guillain-Barré syndrome ( GBS) and explore the relationship between glucose metabolism and severity and prognosis of disease. Methods Records of patients with GBS admitted between January 2004 and February 2017 from Affiliated Hospital of Jiangsu University were evaluated, including antecedent infection, initial symp-toms, cranial nerve palsy, subgroup analysis, Hughes score and Medical Research Council ( MRC) score at nadir and when discharged. Results The incidence of ocular palsy (6. 4％ vs 27. 0％, P=0. 02) and of facial palsy (18. 8％ vs 45. 8％, P =0. 016 ) were both lower in older group, compared to non-elderly group. MRC score at nadir (32 vs 44, P=0. 020), rate of severe type (80. 6％ vs 47. 9％, P=0. 004) and rate of poor prognosis when discharged (67. 7％ vs 29. 2％,P=0. 001) in elderly group were higher than non-elderly group. As to the distribution of subtype in these two groups, no significant difference was found (P=0. 691). Hyperglycemia wasn't prognostic factor of severe type (OR =0. 531,P =0. 321) or poor short-term prognosis (OR=0. 519,P=0. 261). Conclusions The clinical characteristics of elderly patients with GBS are distinct from non-elderly patients. Hyperglycemia wasn 't predictor of severe type or poor short-term prognosis of GBS.

Objective:To study the effectiveness and safety of programmed cell death receptor 1(PD-1) monoclonal antibody combined with chemotherapy in the preoperative neoadjuvant treatment of stage ⅢA non-small cell lung cancer(NSCLC).Methods:A total of 65 patients with stage ⅢA NSCLC who underwent preoperative neoadjuvant treatment in our hospital from January 2019 to October 2020 were selected. According to the preoperative neoadjuvant treatment plan, they were divided into control group(31 cases) and observation group(34 cases). Patients in the control group were treated with albumin-bound paclitaxel and cisplatin for injection, and the patients in the observation group were treated with immunotherapy(carrelizumab/sintilizumab) on the basis of the control group, all underwent 2 cycles of preoperative neoadjuvant treatment. Compared the clinical efficacy of imaging, T lymphocyte subsets, drug side effects, surgical resection rate, major pathological remission(MPR), complete pathological remission(pCR) and postoperative complications of the two groups of patients, and analyzed the factors those affected MPR.Results:The clinical efficacy of PR and ORR of imaging in the observation group was better than that of the control group( P<0.05). The positive rate of CD3 + cells, the positive rate of CD4 + cells, the positive rate of CD8 + cells and the ratio of CD4 + /CD8 + cells in the observation group after treatment were higher than those in the control group( P<0.05). The drug toxicity of the observation group was higher than that of the control group in RCCEP/rash, abnormal thyroid function, and abnormal myocardial enzymes( P<0.05). Compared among the observation group(carrelizumab group/sintilizumab group), the toxicity of carrelizumab group was higher than that of sintilizumab group in RCCEP/skin rash, bone marrow suppression and abnormal myocardial enzymes( P<0.05). The MPR and pCR of the observation group were higher than those of the control group( P<0.05). There was no significant difference in surgical resection rate, surgical methods and postoperative complications between the two groups( P>0.05). The results of univariate analysis showed that ECOG score, pathological type, neoadjuvant treatment plan were related to MPR( P<0.05). The results of binary logistic regression analysis showed that ECOG score and neoadjuvant treatment plan were independent risk factors affecting MPR( P<0.05). Conclusion:PD-1 monoclonal antibody combined with chemotherapy can enable patients to obtain better MPR and pCR, and can improve the immune function of patients. But the side effects caused by immunotherapy drugs are worthy of attention, and the side effects are different between different immune drugs.

Objective:To investigate the preventive and therapeutic effects of resveratrol on lens opacification in diabetic rats and its biological mechanism.Methods:Fifty 8-week-old healthy male SPF grade SD rats were selected and randomly divided into blank control group, model group, gliclazide group, low-dose resveratrol group and high-dose resveratrol group according to their body weight, with 10 rats in each group.The diabetes model was established by intraperitoneal injection of streptozotocin in model group, gliclazide group, low-dose resveratrol group and high-dose resveratrol group.On the third day after modeling, rats in gliclazide group was gavaged with 2 mg/(kg·d) gliclazide suspension, and rats in low-dose and high-dose resveratrol groups were gavaged with 20 and 40 mg/(kg·d) resveratrol, respectively, for four weeks.Rats in blank control group and model group were gavaged with the same volume of normal saline once a day, also for four weeks.After the diabetes model was established, there were 10 rats in blank control group and 9 rats in the other four groups.The fasting blood glucose concentration of the rats was measured with a blood glucose meter.The concentrations of fasting insulin, superoxide dismutase (SOD) 1, SOD2, SOD3, and glutathione peroxidase (GPX1) were determined by enzyme-linked immunosorbent assay.Lens opacification after treatment was observed by slit lamp microscopy.Morphologic changes in lens cells were examined by hematoxylin-eosin staining.Apoptosis of lens epithelial cells (LECs) was detected using TUNEL.The relative expressions of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins in lens tissues were determined by Western blot.The study protocol was approved by the Welfare Ethics Committee of Experimental Animal of Zhengzhou University (No.IACYC2019-02).Results:Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were increased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were decreased in model group in comparison with blank control group, and the differences were statistically significant (all at P<0.05). Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were decreased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were increased in gliclazide group, low-dose resveratrol group, and high-dose resveratrol group compared with model group, and the differences were statistically significant (all at P<0.05). Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were decreased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were increased in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, and the differences were statistically significant (all at P<0.05). The proportions of grade 0, 1 and 2 lens opacities after treatment were 100.00%, 0.00% and 0.00% in blank control group, 0.00%, 66.67% and 33.33% in model group, 77.78%, 22.22% and 0.00% in gliclazide group, 22.22%, 44.44% and 33.33% in low-dose resveratrol group, and 66.67%, 33.33% and 0.00% in high-dose resveratrol group, respectively, with a statistically significant difference ( H=7.514, P<0.001). Compared with model group, lens opacification was less severe in blank control group, gliclazide group, low-dose resveratrol group, and high-dose resveratrol group, with statistically significant differences (all at P<0.05). Lens opacification was less severe in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, showing statistically significant differences (both at P<0.05). Compared with model group, there were fewer abnormal changes of lens cells and sub-organelles in gliclazide group, low-dose resveratrol group and high-dose resveratrol group, and the abnormalities in gliclazide group and high-dose resveratrol group were slighter.Compared with model group, the relative expression levels of Nrf2 and HO-1 were higher in blank control group, gliclazide group, low-dose resveratrol group, and high-dose resveratrol group, with statistically significant differences (all at P<0.05). The relative expression levels of Nrf2 and HO-1 were higher in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, showing statistically significant differences (both at P<0.05). Conclusions:Resveratrol can reduce lens opacification in diabetic rats and its mechanism may be related to the regulation of the Nrf2/HO-1 signaling pathway by exerting antioxidative stress effects.

Objective To investigate the differences in expression of Mohawk (MKX) transcription factors and collagen of types Ⅰ and Ⅲ in anterior cruciate ligament (ACL) gratis between 2 remodeling outcomes under arthroscopy.Methods Enrolled for this study were 17 patients who had undergone arthroscopic single-bundle ACL reconstruction with autogenous hamstring tendons and secondary arthroscopic exploration 48 to 131 months (average,83.1 months) after removal of tibial internal fixator at Department of Sports Medicine,The First Affiliated Hospital to Shenzhen University from March 2017 to December 2017.They were divided into a good remodeling group (11 cases) and a fair remodeling group (6 cases) according to the graft quality under arthroscopy (synovial and vascular coverage,and apparent tension,thickness and retear of the grafts).During the secondary arthroscopic procedures,biopsy of the central ACL grafts was performed.Moreover,normal ACL tissues were harvested from 8 contemporary controls of ＜ 60 years old who underwent total knee replacement.Immunohistochemical assay and quantificational real-time polymerase chain reaction were conducted to detect the expression of transcription factors and collagen of types Ⅰ and Ⅲ in all the samples.Results In the samples from good remodeling and control groups,there were abundant well-arranged collagen fibers of types Ⅰ and Ⅲ and MKX-positive cells;in the fair remolding group,the collagen fibers of types Ⅰ and Ⅲ and MKX-positive cells were much decreased in number and the fibers were not well arranged.The former 2 groups scored in immunohistochemical assay significantly higher than the latter one (P ＜ 0.05).qRT-PCR showed that the expression levels of MKX gene (0.44 ± 0.30),COL1A1 gene (0.52 ± 0.27) and COL3A1 gene (0.60 ± 0.22) in the fair remolding group were significantly lower than in the control group (1.00 ± 0.00,1.00 ± 0.00 and 1.00 ± 0.00) and than in the good remolding group (0.97 ± 0.67,0.99 ±0.38 and 1.00 ± 0.35) (P ＜ 0.05).Conclusion Good remodeling ACL grafts with histological maturation under arthroscopy are more similar to normal ACL than fair remodeling ACL grafts in expression of MKX transcription factors and collagen of types Ⅰ and Ⅲ.

Objective:To summarize the clinical characteristics and therapeutic drug selection of post-transplantation diabetes mellitus(PTDM)after kidney transplantation in children.Methods:From May 2014 to March 2021, a total of 5 cases(5.38%)of 93 paediatric kidney transplant recipients with a median follow-up period of 34 months were diagnosed with PTDM in our centre.Retrospective data analysis was performed for these 5 paediatric recipients.The characteristics of the disease, treatment data and outcomes were summarized.Among the five paediatric recipients, one was male and four patients were female, ranging the age from 12 to 17 years.All recipients received a tacrolimus-based immunosuppressive regimen with prednisone discontinued no later than 3 months after kidney transplant.Results:The onset of PTDM ranged from 1 month to 46 months(median: 17 months)after transplantation.The blood glucose of two children returned to normal gradually after tacrolimus conversion to cyclosporine, with one of them was given insulin temporarily.Three children received oral hypoglycaemic agents, including one received acarbose, one received metformin, and one received metformin combined with acarbose.After a median follow-up of 6 months, the levels of blood glucose in five children were stable, and there was no significant change in serum creatinine and urine protein.Conclusions:The treatment of PTDM in children should be individualized with considering of age, gender and immunosuppressive regimen. Switch from tacrolimus to cyclosporine is effective. Metformin or other hypoglycemic agentsis helpful when tacrolimus is maintained.

Objective:To summarize the clinical characteristics and treatment of cytomegalovirus(CMV)infection in pediatric kidney transplant patients.Methods:From May 2014 to July 2021, a total of 9 cases(8.65%)of 104 pediatric kidney transplant recipients were diagnosed with CMV infection in our centre.Retrospective data was collected for these 9 paediatric recipients.The clinical characteristics of the disease, treatment data and outcomes were summarized.Results:The median age of the 9 children was 10 years(0.25-15 years), 6 of whom were treated with polyclonal antibody for immunity induction.CMV IgG was negative in 4 children before renal transplantation.Only one patient received anti-CMV prophylaxis.The median time from transplant to the diagnosis of CMV infection was 22(7-15)days.Among the 9 children, 7 had fever, pneumonia and diarrhea, 2 had no typical symptoms, three patients were complicated with viral, bacterial or fungal infections.Acute rejection occurred in 3 patients at the same time as CMV infection or after CMV DNA turned negative.Nine patients were cured and discharged after ganciclovir or valganciclovir treatment.Median time of CMV DNA negative transformation was 32(17-90)days.Conclusions:Pediatric transplant recipients are at particularly elevated risk of CMV disease.Antiviral prophylaxis should be initiated early after transplantation.

Objective    To evaluate the efficacy and safety of programmed cell death receptor 1 (PD-1) inhibitor combined with chemotherapy in the preoperative neoadjuvant treatment of stage Ⅲ non-small cell lung cancer (NSCLC). Methods    The clinical data of 68 patients with stage Ⅲ NSCLC who underwent preoperative neoadjuvant treatment in our hospital from June 2019 to October 2020 were analyzed and divided into two groups according to a random number table. There were 34 patients in the control group including 19 males and 15 females with an average age of 59.41±4.77 years. In the observation group, there were 34 patients including 21 males and 13 females with an average age of 61.15±6.24 years. The patients in the control group were treated with albumin-bound paclitaxel and cisplatin for injection, and the patients in the observation group were treated with carrelizumab on the basis of the control group, and both groups received 2 cycles of preoperative neoadjuvant therapy. We compared the clinical efficacy of imaging, T lymphocyte subsets, drug side effects, surgical resection rate, major pathological remission (MPR), complete pathological remission (pCR) and postoperative complications of the two groups of patients, and analyzed the influencing factors for MPR. Results    The objective response rate (ORR) of imaging in the observation group (70.6%) was higher than that in the control group (38.2%, P<0.05). The positive rate of CD3+ cells, the positive rate of CD4+ cells, the positive rate of CD8+ cells and the ratio of CD4+/CD8+ cells in the observation group after treatment were higher than those in the control group (P<0.05). The drug toxicity of the observation group was higher than that of the control group in the reactive cutaneouscapillary endothelial proliferation (RCCEP)/rash, abnormal thyroid function, and abnormal myocardial enzymes (P<0.05). The MPR (66.7%) and pCR (51.9%) of the surgical observation group were higher than those of the surgical control group (MPR: 19.2%, pCR: 7.7%, P<0.05). There was no statistical difference in surgical resection rate and postoperative complications between the two groups (P>0.05). Univariate analysis showed that ECOG score, pathological type, neoadjuvant treatment plan and surgical resection were related to MPR (P<0.05). The results of binary logistic regression analysis showed that Eastern Cooperative Oncology Group (ECOG) score and neoadjuvant treatment plan were independent risk factors for MPR (P<0.05). Conclusion    The clinical efficacy of PD-1 inhibitor combined with chemotherapy in the preoperative neoadjuvant treatment of stage Ⅲ NSCLC patients is definite, and it can significantly improve the patients' MPR, pCR and cellular immune function, but the side effects caused by immunotherapy drugs need to be concerned.

Objective:To explore the efficacy and safety of a novel Tripterygium preparation (Kunxian Capsules)in kidney transplant recipients developing refractory proteinuria after transplantation.Methods:A total of 59 kidney transplant recipients received regular follow-ups from August 2018 to July 2021.Severity of proteinuria, kidney graft function and adverse effects were retrospectively recorded before and at Month 1/2/3/6 months after Kunxian treatment.They were divided into two groups of effective and void to explore the potential effect-related factors.Results:Six-month treatment was completed in 57 patients except for 2 cases of discontinued treatment due to severe adverse effects.A significant reduction in amount of proteinuria was observed at Month 1 [1.09(0.42, 2.59 g/24 h)vs 1.82(1, 2.7 g/24 h, P<0.01)]and the trend continued during subsequent follow-ups.Twenty-nine patients(50.9%)responded remarkably.Eighteen patients(31.6%)showed no response.The overall effective rate was 68.4%.Inter-group comparison revealed strong correlations between treatment efficacy and baseline serum creatinine levels, early initiation of treatment and symptom duration from onset to treatment.Kidney graft IgA nephropathy demonstrated the highest effective rate(11/13, 84.6%). Furthermore, elevated blood concentration of tacrolimus hinted at a potential drug interaction. Conclusions:Kunxian Capsules is efficacious and safe for renal transplant recipients developing moderate-severe proteinuria and not responding to traditional medications.Early initiation of treatment before graft function decline is recommended.Reducing tacrolimus dose cautiously in advance and monitoring adverse events are also essential.

Objective:To summarize the incidence of acute rejection (AR) after pediatric kidney transplantation (KT) at a single center and examine its impact on graft/patient survival and risk factors for AR.Methods:This is a retrospective cohort study including pediatric recipients who underwent kidney transplantation in past 8 years.After excluding recipients of graft thrombosis within a week post-transplant and lost to follow-ups, a total of 143 cases were ultimately recruited and assigned into two groups of AR (n=29) and non-AR (n=114).Basic profiles of both donors and recipients and graft/patient survival rate were compared between two groups.Relative risk factors for AR episodes were also examined by Logistic regression.Results:Renal grafts for 130/143 cases (90.9%) were harvested from deceased donors and 120(83.9%) cases from children.Twenty-seven transplants (18.9%) were performed in infants and young recipients aged < 3 years.During a median follow-up of 33 months, 34 AR episodes occurred in 29(20.3%) patients.Rate of re-transplantation (27.6% vs. 7.9%), pediatric donor (96.5% vs. 80.7%) and rabbit anti-human thymocyte globulin (rATG) induction (79.3% vs. 36%) were significantly higher in AR group than non-AR group ( P=0.007, P=0.046, P<0.001).Multivariate regression analysis indicated that basiliximab induction caused a significant reduction in the risk of AR incidence as compared with rATG induction (odds ratio 0.13, 95% confidence interval 0.04-0.43, P<0.001).The median time of AR incidence was 1.3 months post-transplantation and 23 episodes (67.6%) were confirmed by biopsy.After anti-rejection treatment, 52.9%(n=18) of the cases achieved a full recovery and 38.3% (n=13) had improved graft function.However, 3 cases (8.8%) developed irreversible graft failure.The 1/3-year graft survival rates were significantly lower in AR group than those in non-AR group (75.3% vs. 95.2%, 68.4% vs. 90.4%, P=0.01), and there was no significant difference in 1-and 3-year patient survival rates between two groups. Conclusions:The incidence of AR is relatively high in pediatric renal transplantation, which has an impact on graft survival.Basiliximab induction can effectively reduce the risk of AR.