Objective Laboratory animal practitionres need further management through technical and capacity training.Methods Relying on the Laboratory Animal Center of University and its highly qualified faculty, a high-quality training base for laboratory animal practitioners has been set up.Results Based on the relevant laws and regulations about experimental animals, appropriate training outlines, different training books and courses were prepared and organized for different trainees.Focusing on the detailed and exact experiments, training base has put emphasis on the practical ability and operational skills.Conclusions How to regulate the training for laboratory animal practitioner by means of base training is discussed and tried out in this paper.

Recent studies found that B cell subsets and their factors have double effects on anti-and aiding schistosome infec-tion. This article summarizes the research progress of positive and negative immunoregulation of schistosome infection involving B lymphocytes antibody and regulatory B cells Bregs relating cytokines IL-10 IL-7 and TGF-β .

Objective To explore the anti-inflammatory and anti-angiogenic effect of miR-15a in diabetic retinopathy(DR).Methods High glucose model(HG)was established by treatment with 25 mmol/L glucose in human retinal pigment epithelial cell line HARPE-19,and HARPE-19 cells treated with 5 mmol/L glucose served as control(LG).HG group and LG group were divided into 4 subgroups:miR-15a mimic group,NC-mimic group,miR-15a inhibitor group and NC-inhibitor group.Cell proliferation ability was detected by MTT proliferation assay.Cell migration ability was detected by cell scratch assay.The relationship between miR-15a and ASM was identified by 3′UTR luciferase reporter assay.miR-15a,ASM,VEGF,IL-1β,IL-6,TNF-α mRNA expression was detected by RT-PCR method.Results In NC-mimic cells and NC-inhibitor cells,the expression of miR-15a in HG group was significantly lower than that in LG group(all P<0.05).The expression level of ASM mRNA in miR-15a mimic group was significantly lower than that in NC-mimic group (P<0.05),the expression level of ASM mRNA in miR-15a inhibitor group was significantly higher than that in NC-inhibitor group (P<0.05).The cell scratch width in miR-15a mimic group was significantly higher than that in NC-mimic group(P<0.05).The cell scratch width in miR-15a inhibitor group was significantly lower than that in NC-inhibitor group(P<0.05).The absorbance value in miR-15a mimic group was significantly higher than that in NC-mimic group(P<0.05),the absorbance value in miR-15a inhibitor group was significantly lower than that in NC-inhibitor group(P<0.05).In NC-mimic cells and NC-inhibitor cells,the mRNA expression levels of VEGF,IL-1β,IL-6 and TNF-α in HG group was significantly higher than those in LG group(all P<0.05).Conclusion miR-15a downexpression induces inflammatory reaction and angiogenesis in retinal cells.miR-15a is involved in the pathogenesis of DR by dual regulation of inflammation and angiogenesis.

Objective To investigate the effect of the increased pulse pressure on cerebrovascular hemodynamic index.Methods From January 2014 to October 2016,the application of the method of cross-sectional study,6 006 consecutive health subjects examined and detected with ultrasound noninvasive cerebral hemodynamic index (CVHI) at the Physical Examination Center,Pu′er People′s Hospital,Yunnan Province were enrolled.In order to control the influence of age and gender on the results,according to age and gender in the increased pulse pressure group (≥60 mmHg),and the same sex and age difference of not more than 1 years old were selected as the inclusion criteria in the normal pulse pressure group (60 mmHg).Finally,1 115 subjects were enrolled as a control group for comparative analysis.The study population was divided into an increased pulse pressure group (n=590) and a normal pulse pressure group (n=1 115) in a mixed manner.The differences of CVHI index and its integral score of both groups were compared.The correlation of the increased pulse pressure level and changes of cerebrovascular hemodynamic accumulative scores were analyzed.Results The minimum velocity,mean velocity,and critical pressure in the increased pulse pressure group were significantly lower than those in the normal pulse pressure group (P<0.05),while the differences of the peripheral vascular resistance,characteristic impedance,pulse wave velocity,dynamic resistance,diastolic blood pressure,and critical pressure were significant higher than those in the normal pulse pressure group.There were significant differences (P<0.01).The integral values of cerebral hemodynamics in the increased pulse pressure group and the normal pulse pressure group were 76±22 and 88±19 respectively.There was significant difference (P<0.01).The proportion of the integral value <75 was 34.4% (203/590) and 15.6% respectively (174/1 115,P<0.01).With the increase of pulse pressure level,the integral value of cerebral hemodynamics showed a significant downward trend,there was a significant negative correlation between the two (r=-0.312,P<0.01).Conclusion The increased pulse pressure may result in the changes of cerebral hemodynamic indexes and its integral values.There is a significant negative correlation between the pulse pressure level and the integral value of cerebral hemodynamics.

Objective To investigate the histopathological characteristics of choroidal melanoma. Methods The histopathological data from 64 patients with choroidal melanoma were analyzed retrospectively. The tumor size and the cytological types were observed and detected. The locations of the tumor were classified according to the involved part invaded by the anterior margine of the tumor, and the degrees of the development of the tumor were graded according to the extent of the outward infiltration of the tumor cells. Results In 64 patients with choroidal melanoma, There were large, medium, and small tumors in 25(39.1%), 31(48.4%), and 8(12.5%) respectively. The spindle cell type was found in 42 patients (65.6%) including spindle cell A and B type in 15(23.4%) and 27(42.3%) respectively; epithelioid and mixed cell type was found in 7(10.9%) and 10(15.6%) respectively; the other types were found in 5(7.8%). Twenty-five cases(39.1%)had no invasion with sclera, 22(34.4%)had but limited to sclera, 12(18.8%)penetrated through sclera and 5(7.8%)had intra-orbit infiltration. Conclusion The histopathological characteristics of choroidal melanoma are multiple, and spindle cell type is the most common one. The choroidal melanoma can easily invade the sclera.

Objective To investigate the effect of dexmedetomidine on postoperative patient-controlled intravenous analgesia (PCIA) with sufentanil in patients with essential hypertension. Methods Sixty ASA Ⅱ or Ⅲ patients with essential hypertension aged 42-63 yr weighing 48-72 kg undergoing hysterectomy were randomly divided into 3 groups ( n = 20 each): control group ( group C) and different doses of dexrmedetomidine groups ( group D1.2 ). PCIA was performed with sufentanil 1 μg/ml + tropisetron 5 μg/ml in 100 ml of normal saline within 24 h after operation (background infusion at 2 ml/h with a bolus dose of 0.5 ml and a 15 min lockout interval). Dexmein group C. Ramsay score was recorded. The number of attempts, consumption of sufentanil, the number of patients who needed nifedipine or ephedrine and side effects such as vomiting and respiratory depression were recoded within 24 h after operation. The level of sedation was evaluated with Ramsay sedation score at 24 h after operation.Results Compared with group C, the number of attempts, consumption of sufentanil, the number of patients who needed nifedipine and incidences of vomiting and respiratory depression were significantly decreased, while Ramsay score was significantly increased in D1 and D2 groups, and the number of patients who needed ephedrine was significanlly increased in group D2 ( P ＜ 0.05). The number of attempts and consumption of sufentanil were significantly decreased, and Ramsay score and the number of patients who needed ephedrine were significantly increased in group D2 compared with group D1 ( P ＜ 0.05). Conclusion Dexmedetomidine can not only reduce the consumption of sufentanil for postoperative PCIA, but also prevent postoperative hypertension from deteriorating in patients with essential hypertension.

Objective To obtain the soluble single-chain fragment V (ScFv)monoclonal antibodies (McAbs) against the SSA antigen epitopes.Methods Three octapeptides (60 000 SSA antigen residues 482-493 termed as P1 epitope, residues 310-323 termed as P2 epitope and residues 230-241 termed as P3 epitope) were synthesized on the lysine frame.The McAbs were panned by coating the corresponding as targets.The specificity, affinity and gene squences of the positive clones were assessed.Soluble single-chain fragment V antibodies special for SSA antigen epitopes were expressed and then identificated.Results After 5 rounds of panning, reactive scFv clones contained full-length scFv antibodies coding regions were obtained,with sufficient affinity and specificity for respective antigen peptides.The absorbance values at 410 nm of the fusion protein of anti-P1-P3 activity with the corresponding peptides were 1.43 ± 0.23, 0.82 ±0.31 and 0.80 ± 0.25, and there was also statistically significant difference in the cross reactions ( P ＜ 0.01 ).Three clones were successfully expressed and then purified by His-bind resin.The activity in vivo of soluble ScFv antibodies was identified to be positive by the indirect immune-fluorescence assay on Hep-2 cells.Conclusion Souble ScFv McAbs against corresponding SSA antigen peptides with high affinity,specificity and activity in vivo were obtained, which are to be competent enough for epitopes expression on the target organs.

Objective To analyze the differences of immune responses against Mycobacterium tu-berculosis antigens induced in two different nonhuman primates and to provide rationales for the selection of suitable animal models for vaccine efficacy evaluation.Methods Expression of functional surface markers including CD69 and HLA-DR, the activation markers on CD4+and CD8+T cells from in rhesus macaques and cynomolgus monkeys were measured by flow cytometry analysis.PBMCs were isolated from rhesus ma-caques and cynomolgus monkeys with Mycobacterium tuberculosis infection and stimulated with PPD and pep-tide pools ( ESAT-6/CFP-10) .Enzyme-linked imunospot ( ELISPOT) assay was performed to detect IFN-γproducing lymphocytes.Results The CD4+and CD8+T cells isolated from rhesus macaques without Myco-bacterium tuberculosis infection expressed higher levels of CD69 and HLA-DR than those from healthy cyno-molgus monkeys (P<0.01).The numbers of IFN-γspot forming cells/106 PBMCs in rhesus macaques with Mycobacterium tuberculosis infection for 10 and 11 months were respectively 3 and 3.5 times higher than that of cynomolgus monkeys upon after the stimulation of PBMCs with PPD.The levels of IFN-γproduction by the cells from rhesus macaque group were also higher than those from cynomolgus monkey group upon after the stimulation of PBMCs with ESAT-6 or CFP-10 peptide pools.Conclusion More IFN-γproducing cells were induced in rhesus macaques than that in cynimolgus monkeys after stimulation with Mycobacterium tu-berculosis antigens.Therefore, the rhesus macaques might be a better animal model for evaluating immune responses induced by Mycobacterium tuberculosis vaccines.

Objective To investigate the feasibility of reconstruction of ureter with tubularized peritoneal free grafts for the treatment of avulsion of ureteral mucosa using animal models.Methods Twelve adult dogs were randomly divided into the reconstruction group(n=6) and the control group(n=6).Firstly,the model of avulsion of ureteral mucosa about 3-5 cm long was made.In reconstruction group,tubularized peritoneal free grafts and ureteral stents were placed in the injured ureters;and in control group,no operation was performed but to place the stents into the ureter.The curative effect was observed by IVU and histological examination 10 weeks after operation.Results In reconstruction group,IVU showed normal size and morphology of the kidneys.There was no hydronephrosis,and no obvious stricture of the part of ureter in which peritoneal free grafts were used as mucosa substitutes.In control group,IVU showed no image or only obscure enlarged outlines of the kidneys,and no image of the ureters.Atresia or severe stricture of the ureters was observed in all dogs in control group;while in reconstruction group,the peritoneal membrane was replaced by integrate transitional epithelium,and no obvious stricture was observed.Subepithelial abundant neovascularization was also seen.Conclusions For avulsion of ureteral mucosa exceeding 3 cm,placing stents only will lead to ureteral stricture or atresia,reconstruction using tubularized peritoneal free grafts as mucosa substitutes is an effective method.

Objective To investigate the diagnostic values of human epididymis (HE4)in lung cancer.Methods 80 patients with lung cancer were the experimental group,30 patients with benign pulmonary disease were the benign control group,and 30 healthy people were healthy control group.The levels of carcinoma embryonic antigen (CEA),cytokeratin protein fragment 21-1 (CYFRA21-1),neuron specific enolase (NSE) and HE4 in serum were detected.Results The levels of CEA,NSE,CYFRA21-1 and HE4 in lung cancer patients were higher than those in both the benign control group and the healthy control group (P＜0.05).The areas (AUC) under the receiver operating characteristic (ROC curve) were 0.870,0.818,0.746 and 0.897 for serum CEA,NSE,CYFRA21-1 and HE4 levels in diagnosis of lung cancer.The levels of CEA and HE4 were higher in patients with adenocarcinoma,the level of CYFRA21-1 was higher in patients with squamous cell carcinoma,the level of NSE was higher in patients with small cell lung cancer (SCLC) (P＜0.05).The detections of CYFRA21-1 (AUC=1.000),CEA (AUC=0.727) and HE4 (AUC=0.622) in serum are favorable for the diagnosis of squamous cell carcinoma,The detections of serum CEA (AUC=0.954) and HE4 (AUC=0.944) levels are favorable for the diagnosis of adenocarcinoma,and the detections of NSE (AUC=0.876) was favorable for the diagnosis of SCLC (P＜0.05).Conclusion The levels of CEA,NSE,CYFRA21-1 and HE4 in serum were abnormal in patients with lung cancer.The HE4 level in the patients was correlated with the pathological types and the metastasis of lung cancer.The detection of serum HE4 could be used in the diagnosis and evaluation of lung cancer.