In recent years,the biological therapy of ovarian cancer has made great progress.Immunotherapy including tumor vaccine and adoptive immunity cell therapy can improve immune recognition capability,while the applications of molecular targeted drugs such as monoclonal antibodies and tyrosine kinase inhibitors for different target spots during the tumor cells growth progresses achieve significant clinical benefit,as well as hormone replacement therapy and Chinese medicine treatment improve the life quality of patients with ovarian cancer to a certain degree.

AIM:To explore the expression of anoctamin 1 (ANO1), one of calcium-activated chloride chan-nels ( CaCCs) , in mouse cardiomyocytes and its functional properties.METHODS:The cardiomyocytes from the myocar-dial tissues of C57BL/6 mice were isolated with enzyme and purified by the differential adherent method.The cells were stained with monoclonal anti-sarcomeric actin and Cy3 to evaluate the purity of the myocardial cells.RT-PCR was used to detect the mRNA expression of ANO1 in the mouse cardiomyocytes.The protein expression of ANO1 in the mouse cardio-myocytes was determined by Western blotting analysis.The fluorescence quenching kinetics experiment was used to identify the ion transport properties of ANO1 in the mouse cardiomyocytes.RESULTS: The results of RT-PCR confirmed that ANO1 was expressed in freshly isolated myocardial cells.The results of Western blotting clearly demonstrated the protein expression of ANO1 in primarily cultured myocardial cells.Fluorescence quenching kinetics experiment on freshly isolated single myocardial cell revealed a pronounced outward rectifying property of the ANO1.The functional properties were simi-lar to the classic CaCCs.CONCLUSION:ANO1 expression was identified in the mouse myocardial cells.The function of CaCCs was generated by ANO1, suggesting that ANO1 is the molecular basis of CaCCs.

Objective To construct the eukaryotic expression vectors of fibroblast growth factor receptor 3(FGFR3) MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN, and to detect their expressions in human chronic myeloid leukemia(CML)K562 cell line.Methods The full-length FGFR3 (fgfr3-WT)and dominant negative FGFR3 (fgfr3-DN)were amplified by polymerase chain reaction (PCR). The two genes were respectively digested with EcoRⅠand BamHⅠ,and then ligated into MSCV/puro to construct the recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN which were tranduced into K562 cells by LipofectaminTM 2000 after PCR,double digestion and DNA sequencing.The expressions of FGFR3 protein in K562 cells were detected by Western blotting and flow cytometry. Results The recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN were amplified by PCR method, and the results showed fgfr3-WT of 2 400 bp and fgfr3-DN of 1 200 bp had been successfully cloned into MSCV-puro vector. The 2 400 bp fragment was oblained after double digestion of recombinant plasmid.The sequencing results showed that the size of fgfr3-WT was 2 400 bp which was the same as the sequence from GeneBank.Fgfr3-DN of 1 200 bp was also in conformity with the expected sequence.Compared with control (K562 MSCV)group,the expression level of FGFR3-WT in MSCV/puro-fgfr3-WT transfection (K562-WT)group was increased to above 10 times.There was high expression of FGFR3-DN in MSCV/puro-FGFR3-DN transfection (K562-DN)group,but there was no expressions in control(K562 MSCV)group and K562-WT group.The flow cytometry results showed that the high expressions of FGFR3-WT were in 57.5% cells in K562-DN group and the high expressions of FGFR3-DN were in 41.5% cells in K562-DN group. Conclusion The K562 cell lines highly expressing FGFR3-WT and FGFR3-DN are constructed successfully.

Objective To evaluate pit pattern analysis for detection of early colorectal carcinoma and precancerous lesions. Methods A total of 162 lesions in 144 patients were examined with magnifying colonoscopy after staining, and their pit patter was analyzed with morphology and pathologic diagnosis. Results With confirmation of pathology, there were 34 non-neoplastic lesions and 128 neoplastic ones, in which 12 were carcinomas. The pit patterns in most non-neoplastic lesions (76. 5%, 26/34) were type Ⅰ or Ⅱ , and those in most neoplastic lesions (96. 1% , 123/128) was type Ⅲ, Ⅳ or Ⅴ. Pit patterns of cancerous lesions were mainly type Ⅴ (75.0%, 9/12), and those of 3 cases of advanced cancers were all type Ⅴ N. Conclusion Pit pattern classification is a very important tool to differentiate between neoplastic, nonneoplastic lesions and early cancer, which helps to decide later therapeutic intervention.

Kidney transplantation has become the optimal treatment for end-stage renal disease. However, the demand for kidney exceeds the available supply. In last years, living-related kidney transplantation has progressively increased because of the less rejection, the higher achievement ratio and so on. From 2007 to 2008 ,our hospital succeed in 196 living-related kidney transplantation based on the previous work.

Objective:To study the clinical significance of early detection of soluble growth stimulating gene 2 protein (sST2) and prealbumin (PAB) in patients with chronic pulmonary heart disease (CPHD) complicated with refractory heart failure.Methods:From September 2017 to June 2019, 112 CPHD patients complicated with refractory heart failure were admitted to Hengshui People's Hospital. The selected patients met the revised guidelines for the diagnosis and treatment of chronic obstructive pulmonary disease (2013 revision) and the cardiac function grade Ⅲ-Ⅳ according to the grading criteria of the New York Cardiology Society. Cardiogenic shock, severe liver and kidney dysfunction, malignant tumors, anemia, and autoimmune diseases were excluded. Patients were divided into the high PAB group (≥200 mg/L) and the low PAB group (<200 mg/L) according to the PAB level on admission. The pulmonary artery systolic blood pressure (PASP), pulmonary artery mean pressure (MPAP), and left ventricular ejection fraction (LVEF) were observed in the two groups before and after the treatment. PAB, total bilirubin (TBIL), hypersensitive C reactive protein (hs-CPR), N-terminal B type brain natriuretic peptide (NT-proBNP) and sST2 levels were detected. Measurement data were expressed in terms of mean ± standard deviation, the counting data were compared using χ 2 test. Correlation analysis was conducted using Spearman correlation test. Results:There were 40 cases in the high PAB group and 72 cases in the low PAB group. There were no statistically significant differences in general data between the high PAB and low PAB groups ( P>0.05). Hospitalization time was significantly different between the two groups ( P<0.05). Before the treatment, there were no significant differences in PASP, MPAP, NT-proBNP and LVEF between the two groups ( P>0.05). High sST2 was significantly different between the two groups ( P<0.05). After the treatment, PASP, MPAP, NT-proBNP and sST2 were decreased in both groups, and the improvement was more obvious in the high PAB group than in the low PAB group, with statistical significance ( P<0.05). Before the treatment, the levels of TBIL and hs-CPR were not statistically different between the high PAB and low PAB groups ( P>0.05). However, the levels of TBIL and hs-CPR were beyond the normal range. After the treatment, TBIL and hs-CPR were decreased in both groups, and there was statistically significant difference between the two groups ( P<0.05). Spearman correlation analysis showed that PAB was negatively correlated with sST2 ( r=-0.778, P=0.001). There was a positive correlation between cardiac function and sST2 ( r=0.569, P=0.034), hospitalization time ( r=0.572, P=0.033) in patients with refractory heart failure. The higher the sST2 of CPHD with refractory heart failure, the longer the patient hospitalization time, and the more serious the heart failure was. The area under the combined ROC curve of PAB and sST2 was 0.756. CPHD patients with refractory heart failure had the greatest predictive value. Conclusion:The combined test of sST2 and PAB can evaluate the condition and outcome of CPHD patients with refractory heart failure, and guide the clinic.

Objective To investigate the effects of danhong injection on the apoptosis of SMMC-7721 heptoma cells.Methods The hepatocellular carcinoma cell line SMMC-7721 was incubated with different concentrations(0,2,4,8 μL·mL-1) of danhong injection, and cell morphology was studied under the microscope. The apoptosis index was detected by flow cytometry at 0,6,12,24 h after cultured with 2 μL·mL-1danhong injection. Cell proliferation was measured by MTT assay;Gene expressions of p53,Bax and Bcl-2 were detected by RT-PCR at different drug concentrations. Results The morphology of the hepatoma cells changed obviously,and the apoptosis index increased by danhong injection in a dose-dependant manner. The danhong injection decreased gene expressions of Bcl-2,and obviously increased p53 and Bax. Conclusion Danhong injection can dose-and time-dependently promote apoptosis of SMMC-7721 cells.

Objective To identify and rename one strain stored in National Center for Medicine Culture Collections ( CMCC) and used for tracheitis vaccine production. Methods The test strain CMCC (B)29108 and the type strain DSM30007T were cultured on NA medium. Characteristics in morphology, physiology, biochemistry and fatty acid profile were compared between the two strains. Phylogenetic analysis was based on 16S rRNA and rpoB gene sequences, together with the DNA-DNA hybridization assay. Results A Comparative analysis of a partial sequence of the 16S rRNA gene sequence revealed that the CMCC( B) 29108 strain was closed to the Acinetobacter species and showed the highest similarity with the type strain Acinetobacter baumannii DSM30007T. Moreover, the CMCC(B)29108 strain was highly similar to type strain DSM30007T in morphology, physiology, biochemistry and fatty acid profile. On the phylogenetic tree based on 16S rRNA and rpoB gene of all Acinetobacter members, the CMCC(B)29108 strain steadily clustered into one independent branch only with the DSM30007 T strain with a DNA-DNA hybridization value of 100%. Conclusion The CMCC(B)29108 strain that is one of the strains used for the production of tracheitis vac-cine should be assigned to the species of Acinetobacter baumannii based on its phenotypic, phylogenetic and chemotaxonomic characteristics.

Objective To investigate the expression of anoctamin 1 in Chinese hamster ovary cells (CHO)and to analyze the functional properties of its ion channel,and to provide experimental basis for study on the physiological function of calcium-activated chloride channel.Methods The whole sequence of anoctamin 1 was obtained by PCR technique and was subcloned into pcDNA3.1 to construct the expression vector pcDNA3.1-anoctamin 1 was transfected into CHO by liposome-mediated transfection and the CHO stably expressing anoctamin 1 were aquired by selection with zeocin. The expression and distribution of anoctamin 1 in CHO were measured by RT-PCR technique and inverted fluorescence microscope.The functional properties of anoctamin 1 were measured with halide-sensitive fluorescence proteins YFP-H148Q/I152L.The PBS buffer solution with calcimycin and high concentration of iodine ion was used as experimental group,andthe PBS buffer solution without calcimycin and high concentration of iodine ion was used as control group.Results The results of digestion and sequencing confirmed that anoctamin 1 was cloned into pcDNA3.1 by electrophoresis and blast. The results of RT-PCR and inverted fluorescence microscope indicated that anoctamin 1 was expressed in CHO. The results of I- influx as measured by halide-sensitive fluorescence proteins YFP-H148Q/I152L showed that anoctamin 1 had the more functional properties of trans-epithelial transporting I-,and the transporting speed in experimental group was higher than that in control group (P<0.05).Conclusion Anoctamin 1 can be expressed highly in the CHO;Anoctamin 1 expressed in CHO has the characteristics of calcium-activated chloride channel.

BACKGROUND:Silver nanoparticles (AgNPs) show strong antibacterial effect and are not easy to have drug resistance. But the antibacterial mechanisms of AgNPs have not been wel developed.
OBJECTIVE:To explain the antibacterial mechanisms of AgNPs.
METHODS:We investigated the influence of Ti, TiO2 and TiO2 containing AgNPs onEscherichia coliand Staphylococcus aureus by bacterial inhibition ring test. Escherichia coli was cultured in LB liquid medium with 0, 5, 10 mg/L AgNPs. We measured the absorbance value of bacterial culture. DNA gel electrophoresis was used to study the effect of AgNPs onEscherichia coliDNA. Then we researched the character of apoptosis on Escherichia coli by Annexin V and PI staining, using flow cytometry.
RESULTS AND CONCLUSION:The inhibiting effect of Ti and TiO2 onEscherichia coliandStaphylococcus aureus was not obvious. But the inhibition rings of TiO2 containing AgNPs to bacteria appeared. The absorbance value of Escherichia coliculture was reduced whenEscherichia coliwas co-cultured with AgNPs. And this decrease tendency was in direct proportion with AgNPs concentration. AgNPs reduced the amount of DNA of Escherichia coli and this tendency was directly proportional with AgNPs concentration. TheEscherichia coli apoptosis rate induced by AgNPs was increased and this tendency was positively correlated to the AgNPs concentration. These results indicate that AgNPs can induce bacterial apoptosis to influence the growth of bacteria.