Objective To establish a rapid and accurate method, and to determine the density of mice and rats sperm with enzyme-labeled instrument.Methods ①The optimal wavelengths and the regression equation set up: After six Kunming mice and six Sprague-Dawley rats were sacrificed, the left epididymis was separated and fully cut up in phosphate buffer saline.With water bath, the sperm were fully dissociated.Using the enzyme-labeled instrument to detect the wavelength absorbance respectively under different wavelength and fitting absorbance curve.The best wavelength will be the most close to 1 of the correlation coefficient ( R2 ) and the standard deviation of minimum.After ten Kunming mice and ten Sprague-Dawley rats were sacrificed,the sperm suspension of different concentration gradient were got.The regression equation of the sperm density and absorbance was established by using enzyme-labeled instrument and haemocytometer.②The test of sperm absorbance stability: Mice and rats,six respectively,were used to make the sperm suspension.Samples were put in room temperature (25℃) or 37℃water bath continued,and after water bath about 0,20,30,40,50, 60 min,the change of absorbance was recorded.③The regression equation verification:The mice were administrated orally with 20% ethanol solution for 30 days to make oligospermia.In order to verify the new method, two different method were used to get the sample sperm.Results The optimal absorbancy-sperm density curve could be established at 380 nm.The means of KM mice sperm count ( x1 ) and absorbance ( y1 ) are showed to be the linear function,and the linear regression equation is y1 =2 ×10 -9 x1 +0.0648, R2 =0.9743.The means of SD rat sperm count ( x2 ) and absorbance (y2) are showed to be the linear function,and the linear regression equation is y2 =5 ×10 -9x2 +0.0621,R2 =0.9940.SD rat sperm suspension liquid after 60 min in water bath, absorbance value at 0 min significantly decreased(P<0.05), but at room temperature after 40 min significantly increased ( P<0.05); KM mice sperm suspension in the water bath and under the condition of normal temperature after 50 min, compared with the 0 min, absorbance value increased significantly(P<0.05).Compared with control group, sperm density of ethanol oligozoospermia group by enzyme standard detector and standard curve calculation were significantly decreased ( P <0.05 );compared with absorbancy-sperm density equation, determination of ethanol oligozoospermia group of sperm density by cell counting plate method had not significant difference.The results suggested absorbancy-sperm density equation could effectively detect the reduction of the mice sperm in oligospermia.Conclusion Using enzyme-labeled instrument to set up the curve of absorbancy-sperm density equation can estimate the sperm density of mice and rats rapidly and exactly.

AIM To speculate the hypoglycemic mechanism for rats with type 2 diabetes by exploring the therapeutic effects of leaf aqueous extract from Cyclocarya paliurus on liver insulin receptor (InsR) and insulin receptor substrate 2 (IRS-2).METHODS The diabetic rat model was established through intraperitoneal injection of streptozotocin and fed with high-fat diet.The moleled rats were equally assigned into the control group and leaf aqueous extract from Cyclocarya paliurus group (extract group).After the test extract was orally administrated for four weeks,body weight,urine output,food intake,water intake and fasting blood-glucose (FBG) were measured,and the levels of serum insulin,InsR and IRS-2 mRNA in liver tissue were investigated in rats.RESULTS Compared with the control group,the extract group showed a reduction in urine output,food intake,water intake,FBG and insulin levels.Meanwhile,the rats' body weights in extract group were presented a trend to increase.The gene expressions of InsR and IRS-2 in liver tissue were up-regulated.Moreover,the insulin sensitivity was improved.CONCLUSION The leaf aqueous extract from Cyclocarya paliurus can reduce FBS,improve insulin sensitivity,which may be associated with the increase of InsR and IRS-2 gene expression in liver tissue.

Background: The testis has been reported to be a naturally O 2 -deprived organ, dimethyloxaloylglycine (DMOG) can inhibit hypoxia inducible factor-1alpha (HIF-1α) subject to degradation under normal oxygen condition in cells. Objectives: The objective of this study is to detect the effects of DMOG on the proliferation and differentiation of spermatogonial stem cells (SSCs) in Bama minipigs. Methods: Gradient concentrations of DMOG were added into the culture medium, HIF-1αprotein in SSCs was detected by western blot analysis, the relative transcription levels of the SSC-specific genes were analyzed using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Six days post-induction, the genes related to spermatogenesis were detected by qRT-PCR, and the DNA content was determined by flow cytometry. Results: Results revealed that the levels of HIF-1α protein increased in SSCs with the DMOG treatment in a dose-dependent manner. The relative transcription levels of SSCspecific genes were significantly upregulated (p < 0.05) by activating HIF-1α expression. The induction results showed that DMOG significantly increased (p < 0.05) the spermatogenesis capability of SSCs, and the populations of haploid cells significantly increased (p < 0.05) in DMOG-treated SSCs when compared to those in DMOG-untreated SSCs. Conclusion We demonstrate that DMOG can promote the spermatogenesis activity of SSCs.

One hundred ten patients with Parkinson′s disease(PD)and 110 age-matched healthy controls were enrolled in the study.The non-motor symptoms(NMS)were assessed with Non-motor symptoms of Parkinson′s disease questionnaire(PDNMSQ30).The PD patients were classified as gait abnormality posture type(PIGD, n=73)and tremor type(TD, n=37).The non-motor symptoms were compared between two groups.Results showed that the NMS in PD patients was higher than that in healthy controls[(35.2 ±3.0)vs.(6.0 ±2.0)symptoms, t=6.43, P=0.01].The NMS score of PIGD group was higher than that of TD group[(28.9 ±1.1)vs.(6.2 ±0.7)symptoms, t=2.73, P<0.01]; Mini-mental state examination(MMSE)and Montreal cognitive assessment scale(MoCA)scores of TD group were significantly higher than those of PIGD group(t=11.52,P<0.01; t=8.79,P<0.01).The results indicate that for PD patients with different motor types,their NMS are different,and PD patients with PIGD have more axial involvement,and more NMS.

Objective To investigate the clinical and histopathological features of cervical basal squamous cell carcinoma (BSCC). Methods A retrospective analysis of 10 cases of cervical BSCC was carried out. The clinical data and all the pathological sections were reviewed, the related immunohistochemical results were statistically analyzed, the clinicopathological features were analyzed, and then followed the prognosis. Results (1) Clinical features:the median onset age of BSCC in cervix was 51 years old (ranged 35-69 years old), 5 of them were postmenopausal women. Vaginal bleeding was often seen in clinic (7 cases). Of the 10 cervical BSCC patients, 5 tested HPV types. All of them were HPV positive, including 2 cases of HPV 16 positive and 1 case of high-risk HPV positive. At the time of colposcopy, 3 cases showed exogenous nodular mass, 3 cases showed endogenous infiltrating mass, and 4 cases had unclear type of mass.(2)Treatment:of the 10 patients, 8 underwent hysterectomy+bilateral adnexal excision+pelvic lymphadenectomy, of which 6 underwent radiotherapy or chemotherapy after operation. Radiotherapy and chemotherapy were performed only in 2 cases. (3) Pathological features: histologically, the tumor cells were nests and stripe like growth, which were composed of basal like tumor cells. The cells had obvious heteromorphosis, less cytoplasm, deep dyed nuclei and common nuclear mitosis, and there were often palisade like structures around the cell nests, and some cells in the center of the cell nests were found to have acne like necrosis. It could be mixed with normal squamous cell carcinoma and squamous epithelial lesion. Among the 10 patients, 6 had immunohistochemical results. BSCC mainly expressed p16 and squamous cell markers such as p63, cytokeratin (CK) 5/6 and p40 protein, the positive expression rates were 3/3, 3/3, 2/2 and 3/3, respectively.A few expressed CK7 protein, and the positive expression rate was 1/3. (4) Prognosis:follow-up time ranged from 1 week to 64 months, and 2 cases were lost to follow-up. Among the 8 follow-up patients, 3 had iliac bone, lung or skin metastasis, and 5 had no recurrence or metastasis during the follow-up period. Conclusions BSCC of cervix is a rare malignant tumor of cervix associated with high-risk HPV infection, p16 is more positive. The treatment is similar to that of normal cervical squamous cell carcinoma. Surgical resection and radiotherapy and chemotherapy are the most effective methods according to the clinical stage . At present, the disease is considered to be highly aggressive and the poor prognosis.