Objective To investigate effect of emodin on cell membrane, protein and nucleic acid synthesis of MRSA41577, and systematically investigate the anti-bacterial mechanism of emodin.Methods TTC assay was used to detected the anti-bacterial activity of emodin on MRSA 41577. Conductivity and macromolecular were detected to investigate the effect of emodin on MRSA41577 cell membrane .SDS-PAGE was used to detect the effect of emodin on the soluble protein synthesis.DAPI staining was used to detect the effect of emodin on nucleic acid synthesis.UV-visible spectrophotometric was used to detected the interaction between emodin and DNA.ResuIts Emodin has significant inhibitory activity on MRSA41577, and the minimum inhibitory concentration was 8μg/mL.After treated with 8 μg/mL emodin for 6h, compared with control group, the macromolecular and conductivity improved (71.48 ±0.026)% (P<0.01) and (2.39 ±0.102)%(P<0.05), sepreatly.Compared with control, after treated with 8μg/ml emodin for 16h,the protein reduced 32.8%, and the contents of DNA and RNA reduced (4.82 ±1.06)%(P<0.05,) and (6.67 ±0.36)%(P<0.053).The UV-visible spectrophotometric results indicated that emodin could integrate with DNA through hydrogen bond.ConcIusion The anti-bacterial mechanism of emodin mainly through damage the cell membrane , inhibit the replication and transcription of DNA through hydrogen bond , inhibit the synthesis of protein, and thus inhibit the biological function of bacteria.

Objective: To develop Epstein-Barr virus(EBV)-transformed human peripheral blood B cell lines from healthy volunteers of type B.Methods: B lymphocytes from healthy volunteers of type B capable of producing anti-A antibodies were transformed by EBV,while Cyclosporine A(CsA),as an immunosuppressive agent that could selectively inhibit T-lymphocytes and protect B-lymphocytes from regression,was used in the experiment.Then,the supernanant of the cell culture medium was tested with red blood cells of type A,B and O by agglutinin assay.Results: Twelve of the15 EBV-transformed B lymphocyte cell lines were established,with a success rate of 80%,while 9 human B lymphocyte lines secreted anti-A antibodies.Conclusion: Human B lymphocyte lines secreting antibodies to A antigens were successfully developed,which helps further studies of human blood specific monoclonal antibodies.

Objective: To construct a phage display library of human single-chain Fv antibodies against blood group Rh(D) substance. Methods: Combining phage display library techniques, isolated total RNA from B lymphoblastoid cell lines secreting anti-Rh(D) antibodies was used for the synthesis of the first strand of cDNA, V_ H and V_ L genes were amplified by 2nd PCR and linked together by splicing overlap extension (SOE) with the use of a (Gly_ 4Ser)_ 3 linker. The resulted scFv genes were then cloned into pCANTAB5E vectors and displayed on the phage. Phage clones were selected using intact red cells as a source of antigen. After 4 rounds of "binding-elution-enrichment", each clone was assayed for specificity by Dot ELISA. Results: A phage antibody library, with the sink size being 1.2?107, was obtained. The percentage of full-length scFv gene inserted into phage DNA was 0.80. Rescued by helper phage, a phage scFv library with titer of 3?108 pfu/ml was established. Specific phages with scFv were acquired after 4 rounds of panning, one clone exhibiting specific binding to Rh+ cell was identified by Dot ELISA. Conclusion: A strategy for construction phage antibody library by means of phage display technique was practicable, which would be useful in screening engineered antibodies against human Rh (D) blood group substances.

Objective To evaluate the effect of oxidative injury induced by peroxide oxidase on Klotho expression in mouse renal tubular epithelial cells (TCMK-1) and to explore the possible pathway.Methods TCMK-1 cells were exposed to H2O2 of different concentrations.Reactive oxygen species (ROS) was examined byflow cytometrry.Cell viability was assessed by CCK-8.Cell apoptosis was evaluated by flow cytometry and Hoechst 33258 staining.The expression of Klotho,apoptosis-associated proteins and anti-oxidant enzymes were determined by Western blotting.Results Compared with control group,after H2O2 stimulating TCMK-1 cell,ROS was dramatically elevated (all P ＜ 0.05) and the expression of anti-oxidant enzymes,SOD2 and CAT went down (all P ＜ 0.05);the expression of Klotho was inhibited (all P ＜ 0.05);cell viability of TCMK-1 cells was decreased (all P ＜ 0.05) in a dose-dependent manner (0.3 to 0.9 mmol/L);cell apoptosis was significantly increased in TCMK-1 cells following the concentration of H2O2 (all P ＜ 0.05);Bax/Bcl-2 and the phosphororation of JNK and p38 were obviously elevated in TCMK-1 by H2O2 induction (all P ＜ 0.05).Conclusion Oxidative injuries induced by H2O2 significantly suppresses the expression of Klotho in TCMK-1 cells.And cell apoptosis was increased,p38 and JNK pathway was activated.

Objective Using an adenoviral vector , the wild-type PTEN gene was transduced into activated hepatic stellate cell (HSC) in vitro and the phosphorylation status of Akt were investigated. Methods The wild type PTEN gene was transduced into activated HSC in vitro mediated by adenoviral vector. The expressions of PTEN and total Akt in HSC were measured by Western blot and Real-time fluorescent quantitation PCR. And the expressions of phosphorylated Akt (Thr308) in HSC was determined by Western blot. Results The data showed that exogenous wild type PTEN gene was successfully transduced and expressed in activated HSC in vitro. The over-expression of wild type PTEN resulted in the significant down-regulated expression of phosphorylated Akt (Thr308) in activated HSC (P < 0.01). But no significant defferences were found in the expression of total Akt in activated HSC at both transcriptional and translational levels(P>0.50). Conclusions The overexpression of wild-type PTEN can negatively regulate PI3K/Akt signaling transduction by inhibiting the phosphorylation of Akt in activated HSC in vitro.

Objective:To explore the effect of intragastric administration of diphenoxylate on inducing epilepsy in mice, as well as the changes in neurobehavioral and hippocampal neurons in mice.Methods:Forty C57 male mice aged 2-3 weeks were selected and divided into control group and diphenoxylate group using random number table method, with 20 mice in each group. The mice in the diphenoxylate group were given diphenoxylate (200 mg/kg) by gavage once a day for 14 consecutive days, while the control group mice were given an equal volume of 0.9% sodium chloride solution every day. After each gavage, the seizure status of mice within 2 hours was observed and the mice were graded based on the Racine score. Open field test, elevated cross test, and Morris water maze test were used to observe the neurobehavioral activities of mouse.A digital electroencephalogram machine was used to monitor the epileptic seizures of mice induced by diphenoxylate.Nissl staining was used to observe the damage of hippocampal neurons in mice.SPSS 25.0 software was used for statistical analysis of the data, and independent sample t-test was used for pairwise comparison. Results:The Racine grading results showed that the mice in diphenoxylate group exhibited grade 2 and 3 seizures at 1 hour after gavage. The EEG monitoring results showed that compared with before gavage, the frequency and amplitude of brain waves of mice in diphenoxylate group increased.In the open field test, the residence time in the central region of mice in diphenoxylate group was significantly lower than that of the control group ((12.21±3.37)s, (17.05±4.34)s, t=3.29, P<0.01). In the elevated cross test, the residence time in the open arm of mice in diphenoxylate group was significantly lower than that of the control group ((17.36±5.41)s, (26.70±9.06)s, t=3.31, P<0.01). In the Morris water maze test, the residence time in the platform quadrant of diphenoxylate group was significantly lower than that of the control group((22.08±6.76)s, (27.64±4.60)s, t=2.54, P<0.05). The residence time and the number of stays in the platform area of diphenoxylate group were both significantly lower than those of the control group(both P<0.05). Nissl staining showed that the number of hippocampal neurons in the CA3 region of mice treated with diphenoxylate was significantly lower than that in the control group((135.67±4.59), (140.67±2.73), P<0.05). Conclusion:Excessive diphenoxylate can induce seizures in mice, and the mice exhibit increased anxiety-like behavior and decreased spatial learning and memory abilities.

Objective:To investigate the role of TcpC in uropathogenic Escherichia coli (UPEC)-induced cystitis in mice and to preliminarily analyze the pathogenic mechanism. Methods:C57BL/6 mice were injected with 10 9 CFU wild-type UPEC CFT073 (CFT073 wt) or tcpc gene-deleted mutant (CFT073 Δ tcpc) from urethra into bladder to construct the mouse model of cystitis. The mice were sacrificed 3 d after infection and the bladders were taken to observe the gross pathological changes. Histopathological changes in bladder tissues were observed after HE staining. Immunohistochemistry was used to detect TcpC in bladder tissues. Bacterial loads in urine samples of UPEC-infected mice were counted by tenfold dilution method, and the presence of tcpc gene in the genomic DNA of bacteria from the bladder and urine samples of CFT073 wt-infected mice was measured by PCR. Real-time quantitative RT-PCR (qRT-PCR) and Western blot were performed to detect the expression of TcpC at mRNA and protein levels in macrophages after CFT073 wt infection. The influence of UPEC strains on the activation of NF-κB signaling pathway in macrophages were determined by Western blot. The levels of proinflammatory factors and the bacterial and cell activity after infecting macrophages with UPEC strains were detected by ELISA, laser confocal microscope and fluorescence microscope, respectively. Results:Compared with the mice with CFT073 Δ tcpc infection, CFT073 wt-infected mice had significantly enlarged bladder and severe neutrophil infiltration and abundant TcpC in bladder tissues. The number of bacteria in the urine of CFT073 wt-infected mice was significantly greater than that of the CFT073 Δ tcpc group. PCR results showed that the bacteria in bladder or urine were CFT073 wt. The expression of TcpC at both mRNA and protein levels in macrophages increased significantly after CFT073 wt infection. Moreover, in CFT073 wt-infected macrophages, the expression of IκBα was promoted and the phosphorylation of p65 and the production of proinflammatory factors were suppressed. TcpC was instrumental in the survival and invasion of CFT073 wt in macrophages. Conclusions:TcpC expression increased significantly in mice with CFT073 wt-induced cystitis. TcpC inhibited the activation of NF-κB signaling pathway and the production of proinflammatory factors in macrophages to improve the survival rate of CFT073 wt, which was closely related to the pathogenesis and immune evasion of UPEC.

Objective: To investigate the development status and risk factors of infants and toddlers in rural China. Methods: In this cohort study, 603 infants (6-12 months of age, Phase Ⅰ) in the rural areas of QinLing-Bashan (Qin-Ba) in Shaanxi were recruited in the control group that received no intervention from April 2013 to October 2015. Three follow-up visits were performed every six months (Phase Ⅱ(12-18 months of age), Phase Ⅲ (18-24 months of age) and Phase Ⅳ(24-30 months of age)). In all the 4 phases (Ⅰ-Ⅳ), general data of the children and the families were collected by questionnaires, early childhood growth and development were assessed by door to door visits, children's hemoglobin levels were determined by laboratory tests, and the cognitive and motor development screening was conducted by the Bayley Scales of Infant and Toddler Development. Logistic regression was used to analyze the risk factors affecting the development of infants and toddlers in rural areas and the data were analyzed in terms of risk factors from infants, guardians and family. Results: Phase Ⅱ, Phase Ⅲ and Phase Ⅳ survey recruited 497, 483 and 486 participants respectively. The incidences of cognitive impairment (mental development scores<80) in rural areas of southern Shaanxi were 13.4% (81/603) in Phase Ⅰ(6-12 months), 20.1%(100/497) in PhaseⅡ(12-18 months), 42.9% (207/483) in Phase Ⅲ(18-24 months) and 50.4%(245/486) in Phase Ⅳ(24-30 months) respectively, which showed a significant increase with age (χ²=233.40, P<0.01); the incidences of psychomotor impairment (psychomotor development scores<80) of Phase Ⅰ, Phase Ⅱ, Phase Ⅲ and Phase Ⅳ were 25.0% (151/603), 26.8% (133/497), 8.3% (40/483) and 11.9% (58/486), which showed a significant decrease with age (χ²=87.08, P<0.01). Multivariate logistic regression analysis showed that the leading risk factor of the cognitive development of 24-30-month-old children was the mothers' poor education background (≤9 years of school education) (OR=2.56, P<0.01), and the main risk factors affecting psychomotor development were the mothers' poor education background (≤9 years of school education) (OR=2.64, P<0.05) and growth retardation (OR=2.95, P=0.07). Conclusions The early childhood development (especially cognitive development) in the rural areas of Qin-Ba in Shaanxi of China is not optimistic. More attention should be paid to the early childhood development in rural China, especially to the development of children from the mothers with poor education background.

Objective: To evaluate the success rate during ultrasound-guided percutaneous nephrolithotomy (PCNL) and analyze the reasons of puncture failure. Methods: A retrospective analysis was performed based on the data of 58 patients who underwent ultrasound-guided PCNL by 4 experienced urologists(10 years' experience of PCNL and more than 80 cases per year)in our center from August 2018 to November 2018. Of all the 58 patients, there were 36 males and 22 females (aged from 22 to 73 years) with the mean age of 51 years. The calculi ranged from 9 mm to 93 mm, with the average of 26.5 mm. The separation of renal collecting system ranged from 5 mm to 30 mm, with the average of 15.1 mm. All of the 58 patients underwent one-stage PCNL and the numbers of punctures, the numbers of percutaneous tubes and the reasons for failure were recorded. Results: All percutaneous tubes and surgeries were established and done successfully. Of all the 118 punctures, 74 punctures succeeded by detecting the urine and 68 surgery tracts were established (6 punctures failed because of the dilation). The total puncture success rate was 62.7%(74/118). Of the total 74 successful punctures, 56.8%(42/74)succeeded at the first puncture, 28.4%(21/74)succeeded at the second puncture and 14.9%(11/74)succeeded at least after three punctures. The success puncture rate of the 4 urologists were 59.2%(29/49), 64.1%(25/39), 66.7%(16/24), 66.7%(4/6)respectively, and there were no statistical differences between the puncture success rates of the urologists (P=0.679). Each channel needed 1.7 punctures on average. Fouty-four punctures failed without detecting the urine, including 20 failed punctures because of the puncture tract deviation, 17 punctures without seeing the urine after the core needle being removed, 7 punctures no display on the ultrasound imagine. There were 33 punctures to be tubeless while other 35 indwelling the nephrostomy tubes. Five nephrostomy tubes' position were different with the preoperationally predicted position which means the discrepancy rate was 14.3%(5/35). One patient had complications and classified as Grade II or above on the modified Clavien Grading System of aerothorax. Conclusions The puncture success rate during ultrasound-guided PCNL is not satisfied. The main reasons of puncture failure are the deviation of puncture tract, unclear imaging of puncture tract and other unclear reasons( puncture needle went too deep or superficially or tip of the needle embedded into the stone).

Objective To explore the therapeutic effect of traditional Chinese medicine(TCM)on subsolid nodule(SSN).Methods A practical randomized controlled study method,including 254 SSN patients was adopted.The patients were divided into the TCM(102 cases)and follow-up(152 cases)groups.The follow-up group received regular check-ups in accordance with the guidelines,and the TCM group received TCM syndrome differentiation treatment for 24 weeks.The two groups were compared in terms of the changes in their SSN diameter,SSN number,TCM symptom score,and overall therapeutic effect before and after treatment.Adverse reactions and safety indicators were also recorded.Results The TCM group showed a significantly higher effective rate of treatment(16.7%)than the follow-up group(2.6%)(P<0.01).Compared with their condition before treatment,the TCM group showed no significant changes in their SSN diameter and number but presented considerably reduced fatigue,yellow and red urine symptoms,and overall TCM symptom score(P<0.05).The follow-up group exhibited significantly increased diameter and number of SSN(P<0.01).The follow-up group showed the significantly higher increase in SSN diameter after treatment than the TCM group(P<0.05).Moreover,the follow-up group showed significantly higher fatigue,depression,yellow and red urine symptom scores,and overall TCM symptom score than the TCM group(P<0.05 or P<0.01).Conclusion TCM treatment for SSN has a distinct clinical efficacy,reduces the malignant risk of SSN and improves clinical symptoms of SSN patients,and is safe and feasible.