Objective To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice,construct DXS6804 allelic ladder by molecular clonning and apply them in a population study on the Pumi population in Yunnan,China.Methods PCR was used to produce several different allelic fragments of the locus.After cloning the PCR products,the recombinant plasmids were sequenced.Then we denominated them and used them as template for re-amplification to generate the locus standard ladder.Results The sequencing results confirmed that the size and the construction of the inserts were correct.The genetic polymorphisms of this locus in Yunnan Pumi population of China were studied.Two off-ladder alleles of DXS6804 locus were found.Conclusion This method is of high value for forensic DNA typing to construct standard ladders.DXS6804 is robust for genetic research and forensic application.

OBJECTIVEIn order to adapt the rapid development of modern medicine, this paper is aimed to analyze the application of in vitro diagnostic reagents (IVD Reagents) in hospital management and improve the overall level of hospital management.

METHODSBy groping the management experience of IVD reagents, we discuss the internal hospital management mode of IVD reagents in reality.

RESULTSWith the continuous improvements on the information platform of IVD reagents,we can realize benefit analysis of IVD reagents within the process of management.

CONCLUSIONReasonable management on IVD reagentscan improve the working efficiency in hospitals and provide swifter and better medical service for patients.

Objective To evaluate the changes of serum tumor markers of carbohydrate antigen 125 (CA125),carbohydrate antigen 199 (CA199),carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC) during anti-tuberculosis treatment in patients with active pulmonary tuberculosis and tuberculous pleurisy.Methods This research was a prospective study.Sixty-three patients with active pulmonary tuberculosis and 24 patients with tuberculous pleurisy underwent blood samplings before treatment,2 and 6 months after treatment.Centaur XP chemiluminescent immunoassay was used to test serum levels of CA125,CA199,CEA and SCC.Thirty healthy subjects were included as controls.Student t-test was used to compare continuous variables,and chi-square test or Fisher exact test was used to compare categorical variables.Serial changes of serum tumor markers levels pre-and post-treatment were analyzed by repeated measures analysis of variance.Binary Logistic regression model was used for multivariate analysis.Results The mean serum CA125 level of the 63 patients with active pulmonary tuberculosis pre-treatment was (64.4± 30.4) U/mL,which was significantly higher than that of healthy controls ([12.7±5.5] U/mL,t=11.98,P＜0.01).The mean serum CA125 level decreased to (16.9±6.1) U/mL after 6 months standardized treatment,which was significantly lower than that before treatment (t=12.74,P＜0.01).While compared with healthy controls,the serum level of CA125 in patients who had completed the standardized treatment was not significantly different (t =0.94,P =0.348).When compared with the healthy controls,serum CA199,CEA,SCC levels in patients with active pulmonary tuberculosis before and after treatment showed no statistically significant difference (P＞ 0.05).Univariate analysis and multivariate analysis by Logistic regression analysis showed that bilateral pulmonary tuberculosis (x2 =7.746,P=0.006; OR=6.99,95％CI:1.73-28.22) and cavity pulmonary tuberculosis (x2 =6.254,P=0.012; OR=7.64,95％CI:1.64-35.35) were associated with increased serum CA125 level.The mean serum CA125 level of 24 cases of tuberculous pleurisy pre-treatment was (81.2 ± 37.6) U/mL,which was both significantly higher than that of patients with active pulmonary tuberculosis (t=2.153,P=0.034) and that of healthy controls (t=12.05,P＜0.01).Similarly,CA125 levels sharply decreased to (15.5 ± 7.3) U/mL after 6 months standardized treatment,which was not statistically significant compared with the control group (t=0.450,P=0.652).However,CA199,CEA and SCC levels in tuberculous pleurisy pre-and post-treatment were all not statistically different from those of healthy controls (all P＞0.05).Conclusion Serum CA125 can be used as a marker for assessing the disease progression and therapeutic efficacy for patients with active pulmonary tuberculosis and tuberculous pleurisy.

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.

Objective To observe the effect and safety of Ginkgo biloba extract (EGb) in patients with chronic allograft nephropathy (CAN),and to study the clinical protective mechanism of EGb.Method A prospective,non-randomized,controlled study was conducted on 103 cases of CAN from March 2013 to March 2015.All patients were divided into experimental group (group A,53 cases) and control group (group B,50 cases).The group A was treated with EGb.Patients were followed up for at least 6 months.Before and after treatment,the changes in renal hemodynamic parameters were observed.The biochemical parameters were also observed,including 24-h urinary protein,urinary albumin,serum creatinine (Scr),triglyceride (TG),total cholesterol (TC),estimated glomerular filtration rate (eGFR),platelet count (PLT),fibrinogen (FIB),D-dimer (DD),activated partial thromboplastin time (APTT).The clinical efficacy and safety were analyzed.Result (1) Therewere no significant differences in clinical and biochemical parameters between the two groups before treatment (P＞0.05).(2) After treatment,the systolic peak flow velocity (Vmax) of segmental artery and arcuate artery in the experimental group was significantly higher than in the control group,and the resistance index (RI) in the experimental group was significantly lower than that in the control group,P＜0.05.(3) In both two groups,the 24-h urinary protein,urinaryalbumin,TG,TC and Scr were decreased after treatment (P＜0.05),and eGFR was elevated (P＜0.05).Moreover,the changes in 24-h urinary protein and urinary albumin in the experimental group were more significant than the control group after treatment (P＜0.05).(3) PLT,FIB and DD in experimental group were significantly decreased after treatment,and APTT was increased significantly (P＜0.05).PLT,FIB,DD and APTT had significant change after treatment in the experimental group as compared with control group.(4) There were no significant differences in adverse reactions between two groups (x2 =0.047,P =0.828).Conclusion The therapy of EGb in patients with CAN could reduce urinary protein and improve hypercoagulable state,and had few adverse reaction with good security.

Objective:To research the number and function of monocyte-macrophages in patients with chronic active hepatitis B. Methods:The 51 chronic viral hepatitis B( CHB) patients were selected randomly,which consisted of 20 cases of mild-moderate,31 cases of severe group and 13 cases of healthy controls. PBMCs were separated by percoll. Monocytes were tagged by CD14,the molecules CD80,CD86,HLA-DR and CD163 were detected by flow cytometry which expressed on the surface of PBMCs. Serum cytokine were detected for IL-10, IL-12 and IL-23 by ELISA. The distribution of CD68 was detected in the liver by immunohistochemical staining. Results:The expressions of CD80 for all chronic hepatitis B patients were lower than the controls respectively,no matter mild-moderate or even severe group. Similarly,the HBV patients expressed lower level of CD86 in the peripheral blood mononuclear cells when compared with the control group. Furthermore, there was statistically difference between the levels of CD86 in severe group compared with control group (P<0. 01). As the expression of CD80 and CD86,the levels of HLA-DR in the patents had also declined when compared with controls. While the HLA-DR levels in both the mild-moderate HBV hepatitis groups were statistically significant higher than the severe group (P<0. 01). Different from the above all,the expression of CD163 in all chronic HBV hepatitis was higher than the control group. The CD68 positive cells in chronic HBV patients were observed and infiltrated increasingly in portal area and hepatic lobules (P<0. 05). There were statistically significant differences of IL-10 levels between the mild-moderate group,severe group and the control group,respectively (P<0. 01). Conclusion:Macrophages have participated in the pathological lesions of liver in CHB patients,among peripheral blood mononuclear cells,the phenomena of imbalance between type M1/M2 and polarization to type M2 have been observed,which participated in the development of the chronicity of CHB.

Objective To probe into the content of DNA Topo Ⅱ in osteosarcoma after chemotherapy.Methods 30 patients with osteosarcoma received two courses of chemotherapy treatment before the surgical resection of the tumor tissue.Then immunohistochemistry was used to detect the content of Topo Ⅱ in tissues and detected its relationship in pathology.Results There were 8 out of 30 cases in which Topo Ⅱ was presented positive in osteosarcoma (26.7 ％).The protein content of Topo Ⅱ was unrelated to the patient' s age,gender,degree of tumor malignancy,tumor location and translocation or Enneking staging (P ＞ 0.05),but related to patients survival rate (P ＜ 0.05).Conclusion Patients with lower expression of Topo Ⅱ are more likely to have poor prognosis.

Objective To enhance the yield and product quality of diosgenin extract,the method of stepwise biocatalysis was used.Methods The optimum conditions of slepwise biocatalysis were obtained through orthogonal tests.In qualitative and quantitative analysis of diosgenin extracted through stepwise biocatalysis,IR spectrum and HPLC chromatogram were used.The yield and melting point of diosgenin were taken as investigated standard index to compare this method with others,such as acid hydrolysis,spontaneous fermentation,and enzymatic hydrolysis.Results When cellulase and pectinase compound,amlyse and diastatic enzyme were added in order in the method of stepwise biocatalysis,98% diosgenin from plants was extracted.IR spectrum of the product was in accordance with that of reference substance and the purity of it is more than 95% according to its HPLC chromatogram.Conclusion Compared with other three methods,it can improve the yield and quality with less energy consumption by stepwise biocatalysis.

Objective:To investigate the expression of sal-like 4 (SALL4) gene in children with acute leukemia and analyze its clinical significance. Methods:Real-time PCR and immunohistochemistry were used to detect SALL4 mRNA and SALL4 protein ex-pressions in 50 patients initially diagnosed with acute leukemia and in 15 patients with immune thrombocytopenic purpura (ITP), which served as controls. Changes were detected in SALL4 mRNA expression from preliminary diagnosis and after complete remission of 5 acute leukemia patients. The relationship between SALL4 mRNA expression and clinical indicators was analyzed. Results: SALL4 mRNA expression is higher in initially diagnosed B-ALL [13.89 (1.00-63.15)] and AML [11.12 (2.31-56.59)] than in ITP controls [1.00 (0.29-1.71)] (P<0.01). SALL4 mRNA expression in initially diagnosed T-ALL [1.48 (0.87-4.81)] and in controls showed no significant difference (P>0.05). SALL4 protein expression is in agreement with SALL4 mRNA expression. SALL4 mRNA expression significant-ly decreased in complete remission stage [0.98 (0.22-1.09)] than in acute phase [28.64 (11.20-87.46)] in acute-leukemia patients (P<0.01). High SALL4 mRNA expression level is positively correlated with high white blood cell count, high risk classification, and high minimal-residual disease at the end of induction chemotherapy (r=0.424, r=0.40, and r=0.393, respectively;P<0.05), but not with age, gender, hepatomegaly, splenomegaly, and lymphadenectasis (P>0.05). Conclusion:SALL4 was found to play an important role in pro-moting childhood B-ALL and AML, which promises a new target for monitoring the therapeutic effects and evaluating the prognosis of childhood B-ALL and AML.

BACKGROUND:Skeletal muscle satelite cels are muscle-derived stem cels with proliferation and differentiation potential distributing between the muscle cel membrane and the base film. Studies have shown that skeletal muscle satelite cels are of efficacy and safety, but the survival rate of the transplanted stem cels is very low, which greatly limits the application of skeletal muscle satelite cels. OBJECTIVE: To observe the effects of Fasudil on apoptosis of skeletal muscle satelite cels induced by H2O2. METHODS: Skeletal muscle satelite cels cultured in vitro were randomly divided into three groups including H2O2group, H2O2+Fasudil group (Fasudil group) and control group. Apoptosis rates were observed by flow cytometry. The concentrations of interleukin-4 and tumor necrosis factor-a in each group were detected by ELISA. Western blot was employed to measure the protein level of Bax in each group. RESULTS AND CONCLUSION: Compared with the H2O2group, a significant decrease was found in the apoptosis rate of cels, protein level of Bax, and concentrations of interleukin-4 and tumor necrosis factor-a in the Fasudil group (alP < 0.05). These findings indicate that Fasudil can play anti-apoptosis protection by inhibiting Rho-kinase signaling pathway, which may be related to the reduced expression of Bax.