Abstract:Objective: It has been widely accepted that lymphatic helper T cells in the lymph system have plasticity. Treg cells, as one of the helper T cell phenotypes, can be induced in different cytokine environment and reveal the existence of plasticity. In addition, lymphocyte function associated antigen 1 (LFA-1) play an important role in the function and migration of lymphocyte differentiation. This study was aimed to investigate the effect of LFA-1 on Treg cells plasticity and the possibility mechanism in vitro. Methods: The high purity Treg cells in wild mice and LFA–1-/- mice pre in vitro, and then cultured in completely medium containing IL-6, IL-23, TGF–beta. On the eighth day, the co-expressing of Foxp3 and IL-17 was measured by flow cytometer, and the expression of STAT3 and ROR gamma t was detected by PCR. The levels of IL-10 and IL-17 in supernatant were measured by ELSIA. Results: Both of the wild mice Treg cells and LFA–1-/- mice Treg cells showed the plasticity in inflammatory conditions, but LFA–1-/- mice Treg cells revealed a higher percentage of Foxp3/IL-17 co-expressing, and the expression of STAT3 and ROR gamma t in LFA–1-/- mice Treg was also increased than that in the wild mice Treg cells(P<0.001). Furthermore, both of the two types of cells supernatants showed that IL-17 is higher with IL-10 decreasing than that of treatment，and the change is obvious in LFA–1-/- mice Treg cells(P<0.001) . Conclusion: LFA-1 has the effect on the plasticity of Treg by regulating the expression of STAT3 and ROR gamma t.

Abstract:Objective: To investigate the effects of methamphetamine (Meth) on the protein expression of β-amyloid precursor protein（APP） and p-tau. Methods: Thirty C57BL/6J male mice were randomly divided into control group, saline treatment group and Meth treated group, then the mice were administrated intraperitoneally with 10 mg/kg Meth for 7 days and two times a day. The silver staining of Golgi apparatus was performed to investigate the number of neurons and the dendritic spines, and the pathological protein APP and p-tau were detected by western blot. Results: With the silver staining of Golgi apparatus, it showed that Meth significantly decreased the number of the neurons as well as the dendritic spines. With the Western blot assay, it suggested that Meth obviously up-regulated the pathological protein APP and p-tau expression in a dose- and time-dependent manner（P<0.05）. Intriguingly, pre-incubated with the neural cells with nifedipine, a blocker of the L-type Ca2+ channel, substantially attenuated Meth-mediated up-regulation of APP and p-tau. Conclusion: Meth exposure contributes Alzheimer's disease-like pathological changes, therefore, the present work will provide some supports for Meth-induced cognitive decline.

Abstract:Objective: To explore the effects of FBXO22 silencing by small interfering RNA (siRNA) on invasion and migration of colon cancer cells, and futher to investigate its related molecular mechanism. Methods: The siRNA-Ctrl and siRNA-FBXO22 fragments were transfected into SW620 and HCT116 cells, and the level of FBXO22 was detected by Western bolt. The effects of interfering FBXO22 on cell invasion and migration were analyzed by Transwell assay, and the expression levels of FBXO22, MMP-2, MMP-9 and MDM2 were detected by Western bolt. Results: After transfection with FBXO22 siRNA, the levels of FBXO22 in SW620 and HCT116 cells were significantly decreased, and the abilities of cell invasion and migration were decreased. Meanwhile, silencing of FBXO22 decreased expression of MMP-2, MMP-9, and increased expression of MDM2. Conclusion: FBXO22 can affect cell invasion and migration in colon cancer cells, and the mechanism may be related to the expression of MMP-2 and MMP-9.

Abstract:Objective: To observe the effect of exogenous gastrin-17 on the expression of E-Cadherin and N-Cadherin in human gastric cancer cell line SGC-7901, and discuss the related mechanisms. Methods: SGC-7901 cells were pre-treated with or without YM022 for 1 h and then incubated with G-17 for 30 min to detect the activation of JAK2/STAT3 signaling pathway, for 24 h to detect the expression of E-Cadherin and N-Cadherin, respectively by western blot analysis. SGC-7901 cells were transfected with CCK2R-siRNA or full-length cDNA of human CCK2R (pCMV6-CCK2R), followed by G-17 treatment to detect the expression of p-STAT3, E-Cadherin and N-Cadherin. Cells were pre-treated with AG490 for 1 h or knockdown of STAT3 with siRNA, then incubated with G-17 for 24 h to evaluate the expression of E-Cadherin and N-Cadherin. Results: Western blot assay showed the exogenous G-17 significantly decreased the expression of E-Cadherin and increased the expression of N-Cadherin, meanwhile, activated the JAK2/STAT3 signaling pathway. Specific antagonist or siRNA against CCK2R partly blocked gastrin-induced activation of STAT3 and the expression of E-Cadherin and N-Cadherin, which suggested the effect of gastrin was CCK2R dependent. Specific inhibitor of JAK2/STAT3 signaling pathway AG490 and siRNA against STAT3 partly attenuated the effect of gastrin on the expression of E-Cadherin and N-Cadherin. Conclusion: Gastrin acting on the cholecystokinin2 receptor, down-regulates expression of E-Cadherin and up-regulates the expression of N-Cadherin, via activation of JAK2/STAT3 signaling pathway induced EMT in human gastric cancer cells.

Abstract:Objective: To investigate the effect of Pioglitazone on genes involved in bile acid enterohepatic circulation in C57bl mice. Methods: C57bl mice were randomly divided into two groups, named NC (normal control, n=8) group and M (medicine, n=8) group, respectively. Mice in NC group were fed with chow diet, while mice in M group were fed with both Pioglitazone and chow diet, and the feeding last for 10 weeks. At the end of our experiment, all mice were sacrificed and blood samples,liver and intestine tissues were collected. Fully automatic biochemical analyzer was used for the detection of blood metabolic parameters, and qRT-PCR was used for the detection of gene expression，which were involved in bile acid enterohepatic circulation. Results: Mice in group M displayed an increased biliary BA content. FXR, CYP7A1 and CYP27A1，which play important roles in bile acid synthesis were detected in an induced expression in group M mice. Hepatic bile acid transporters BSEP and NTCP were also detected in an increased expression in group M compared with group NC. Immunohistochemical examination testified an increased protein expression of BSEP in mice liver tissue. ASBT, a gene regulates intestinal bile acid reabsorption, were detected in an increased expression in group M mice. Conclusion: Pioglitazone increased the expression of genes involved in bile acid synthesis, bile acid transport,as well as intestinal reabsorption,which indicates the drug，may play a great role in regulating genes involved in enterohepatic circulation.

Abstract:Objective: To explore the new biomarkers for predicting the side effects of UGT1A1 genotyping directed chemotherapy. Methods: A total of 269 patients were enrolled in the present study. Each patient was determined polymorphism of UGT1A1 gene for genotyping. Serum biochemical markers including alanine aminotransferase (ALT), aspartate aminotransferase(AST), creatinine (Scr) and plasma albumin (ALB) were also analyzed routinely prior to treatment. According to the NCCN(2017V1) guidelines, the differential doses of irinotecan were set according to the genotyping of the patients. The relationships between the side effects and blood biochemical markers were analyzed by Wilcoxon's rank-sum test. Results: The overall toxic effects in patients was significantly reduced under the genotyping directed irinotecan treatment in patients. However, 31 cases (11.5%) were found with severe irinotecan specific toxic effects. Furher analysis indicated that serum AST was associated with toxic effects of irinotecan but with lower predicted value (OR=0.98, 95% confidence interval: 0.97 ~ 1.00, P<0.05; Area under the ROC curve, AUC was 0.6797). Interestingly, low ALB level before treatment was indicated significant correlation with irinotecan induced toxic effects (OR=1.51, 95% confidence interval: 1.34 ~ 1.71, P<0.05). ROC analysis showed that the area under the curve of albumin was reached up to 0.9266. However, pathological type of tumor, gender, age, and other serum biomarker were without obvious correlations with irinotecan induced toxic effects. Conclusion: Low ALB before treatment was significantly associated with irinotecan induced toxic effects in patients and it might be a potential predicting biomarker for UGT1A1 genotyping directed chemotherapy in clinic.

Abstract:Objective：This study aimed to investigate whether acupuncture at ST36 can make effects on the special weapons and tactics (SWAT) trainees’ recovery from special training load. Methods: We recruited 131 SWAT trainees, divided into 5 groups randomly, and then gave acupuncture or Sham needle, give positive language introduction or negative language introduction, thus form four experimental groups: Sham needle high expectations group(n=26), Sham needle low expectations group (n=26), acupuncture high expectations group(n=26), acupuncture low expectations group(n=26) , and we set blank control group (n=27). With the help of Swedish Monark839E power vehicle, progressive load motion test was carried out to increase the gradient 15 W/min until exhaustion.The heart rate(HR), oxygen volume (VO2), respiration rate(RR), rating of perceived exercise (RPE)， systolic blood pressure(SBP）and diastolic blood pressure (DBP） were tested during the training and recovery process. In the six minutes after the exercise, the experienced clinical acupuncture doctor performed Sham needle or acupuncture, leaving the needle for 20 minutes. The acupuncture point was ST36, the position of the Sham acupuncture point is to away from the non-meridian point 1~2 cm.The control group recovered naturally. According to the true or false of acupuncture and expectations of high or low, it formed ABC three contrasts: four experimental groups together versus blank control group, two acupuncture group together versus two sham needle group, the two lower expectations groups together versus two higher expectations.Compare the level of fatigue recovery of above contrasts. Results: The two-factor variance analysis in the three control groups on HR, RPE, RR, VO2 were showed statistical significance (P< 0.05) except SBP and DBP. Conclusion: Acupuncture has three and high psychological expectations to promote the movement fatigue recovery process.

Abstract:Objective: To investigate the correlation between thyroid function within the normal range and bone mineral density as well as trabecular bone score in men of different age groups. Methods: Total of 191 men <60 years old as well as 97 men ≥60 years old, conformed to the inclusion criteria, were recruited. Free triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH) and ordinary biochemistry indexes were measured. Dual energy X-ray absorptiometry (DXA) was used to measure bone mineral density (BMD) of femoral neck, total hip and lumbar spine. Trabecular bone score (TBS) was calculated with TBS iNsight software. Results: In both groups, TBS was positively correlated with BMD of each part tested. FT3 was negatively related with total hip BMD, lumbar spine BMD and TBS in young and middle-aged men. However, in elderly men, femoral neck BMD, total hip BMD and TBS were negatively related to age, which were not associated with thyroid function. Conclusion: There is a good correlation between TBS and BMD. Elevated FT3 level within the normal range was associated with bone loss and bone microstructure destruction in young and middle-aged men. In elderly men with normal thyroid function, age may have more adverse effect on bone mineral density and bone microstructure compared to thyroid function.

Abstract:Objective: To compare the clinical outcomes of patients with normal ovarian function in vitro fertilization and embryo transfer therapy, using clomiphene citrate combined with low dose gonadotropin regimen and conventional antagonist regimen. Methods: A prospective randomized controlled trial was conducted. One hundrend and tweenty couples were selected from September 2014 to December 2015 at the First Affiliated Hospital of NJMU, the normal ovarian function of the woman and the in vitro fertilization embryo transfer therapy. The women were divided into experimental group (clomiphene citrate combination low dose gonadotropin regimen) and control group (conventional antagonist regimen) according to the randomized digital envelope method. Both of the two groups All embryo were frozen and frozen-thawed embryos were transfered, followed-up to live or running out of all embryos of this cycle in two groups, compare the clinical and laboratory results and clinical outcomes of the two groups were compared. Results: The dosage of gonadotropin in the experimental group was lower than that in the control group. The stimulate time of gonadotropin was shorter than that of the control group. There was no significant difference in clinical pregnancy rate and cumulative live birth rate between two groups. Conclusion: The combination of clomiphene citrate and low dose gonadotropin program can achieve the clinical outcome as good as the conventional antagonist regimen in the case of reduced dosage of gonadotropin and shortened stimulation days. It is a new economical, effective and mild ovulation induction program.

Abstract:Objective: The main purpose is to investigate the effect and mechanism of sodium valprate on the transcriptional expression of the human orosomucoid 1-like protein3(ORMDL3). Methods: The effect of sodium valprate on the mRNA expression of ORMDL3 was determined by using semi-quantitative RT-PCR. The effect of sodium valprate on ORMDL3 promoter activity and its mechanism were detected by point mutations and Dual-Luciferase reporter assays. Results: By adding sodium valprate(final concentration of 2 mmol/L) in human embryonic kidney 293 cells (HEK293 cells), it can significantly reduce the expression amount of the human ORMDL3 gene. The luciferase analysis showed that sodium valprate can significantly inhibit the promoter activity of ORMDL3 gene. P300 and CREB binding to the promoter region can drive the ORMDL3 transcription. When sodium valprate act on the plasmids without the binding region of p300 or CREB, there was no significant difference compared with the wild-type plasmid. Conclusion: Sodium valprate can reduce the mRNA level of the human ORMDL3 in HEK293 cells independently of binding with the p300 and CREB in core promoter area of ORMDL3 gene.

Abstract:Objective: To evaluate clinical results and changes in cervic sagittal parameters after anterior cervical discectomy and fusion (ACDF) and to analyze the difference on clinical prognoses among treatment of different surgical segments. Methods: A retrospective study was performed in 52 cases of cervical disc herniation from February 2013 to January 2016 in the First Affiliated Hospital of Nanjing Medical University, who were undertaken by ACDF. Before and after the operations, cervical sagittal parameters were measured in MRI, including C0C2 angle, C2C7 angle, T1 slope, neck tilting and thoracic inlet angle. Therapeutic effects were evaluated by pain visual analogue scale (VAS) and Japanese Orthopaedic Association Scores (JOA). Results: The C2C7 angle and T1 slope were increased with the decreased neck tilting postoperatively. Also,the scores of VAS and JOA were improved after ACDF with a significant difference (P<0.05). Compared preoperative and postoperative results among single-level segment, double-levels segments and multiple-levels segments treatment, there were significant differences in the duration of operation and the amount of bleeding (P value =0.001). However, no statistical difference was found in cervical sagittal parameters changes and clinical improvement rate. Conclusion: Anterior cervical discectomy and fusion for cervical spondylosis comes out with a good therapeutic effect, which could improve the cervical curvature and increase the T1 slope to restore cervical sagittal balance. No significant difference was found in the changes of cervical sagittal parameters for different segements as well as clinical prognoses postoperatively in the treatment of cervical disc herniation.

Abstract:Objective：To investigate the relationship between CD3+ T cells and CD8+ T cells infiltration and atypical hyperplasia in oral lichen planus (OLP) , which will provide a new direction for the clinical diagnosis and immunotherapy of OLP. Methods：Immunohistochemical semi-quantitative method was used to detect the CD3+ T cells and CD8+ T cells infiltration in 79 cases of OLP patients, meanwhile 10 cases of normal oral mucosa as a control. The relationship between these T cell infiltration and atypical hyperplasia in OLP was analyzed by statistical methods. Results：CD3+ T cells and CD8+ T cells were negatively expressed in normal oral mucosa, but were positively expressed in 79 cases of OLP. The infiltration of CD3+ T cells and CD8+ T cells had no relationship with gender, age or the location of OLP, but they were closely correlated with the atypical hyperplasia of OLP（P<0.001 and P=0.002, respectively）, the infiltration of CD3+ T cells were positively associated with the infiltration of CD8+ T cells（r=0.498，P<0.001）. Conclusions：The infiltration of CD3+ T cells and CD8+ T cells is closely related to epithelial dysplasia, which affects the occurrence and development of OLP.

Abstract:Objective: To use of modified Politzer device in the management of chronic otitis media with effusion(OME) in children and evaluate the clinical efficacy. Methods: Forty-three Children diagnosed with bilateral chronic OME and hearing loss were randomly divided into A and B groups. Tweenty-two Children in group A, aged 4~11 years started as the treatment group underwent modified Politzer device for inflation. Another 21 Children in group B, aged 5~12 years were included as the control group underwent careful observation. After 4 weeks, a cross-over was performed. Both groups underwent otoendoscopy, tympanometry and audiometry at inclusion and after 4 and 8 weeks for evaluating the treatment efficacy. Results: In group A, the middle ear pressure and pure tone threshold were normalized in 20(45% )and improved in 14(32%) ears after 4 weeks treatment. While in B group, 6(14%) ears were normalized and 4 ears were improved(10%) . Statistically significant differences were demonstrated between children with modified Politzer device for inflation and children with observation. After cross-over , 17(40%) normalization and 13(31%) improvement of ears were achieved in B group, and 24(55%) normalization and 14(32%) improvement of ears were achieved in A group. Statistically differences were demonstrated between two groups (P＜0.05). There were 8(19%) children were submitted to grommet surgery after 8 weeks treatment. in two groups. Conclusion: Efficiency were demonstrated in children with chronic OME inflated with Modified Politzer. The earlier treatments were given, the better effects could be achieved. It can reduce the need for surgery.