Protein protein binding is the basis of virus and host cell interactions. With the application of technology of studying of protein interactions, more knowledge of replication and pathogenesis of hepatitis C virus (HCV) could be acquired. Non structure protein NS5A is one of the important regulatory factors in virus replication , transcription and signal transduction, but there are controversy in effect on HCV pathogenesis and resistance to interferon ?(IFN?). In order to describe the relationship between NS5A and host proteins, we use yeast two hybrid system 3 to screen the gene encoding proteins that could interact with NS5A from human hepatocyte library. Thirty five clones were obtained including apo A1, apo A2, apo B100, haplotype mitochondrion complete genome, phosphatidic acid phosphatase type 2B, albumin similar to tumor endothelial marker 5 precursor, matrix metalloproteinase 14, and three of the Homo sapiens hypothetical proteins. The study paved a way for further studies on the pathogenesis of HCV NS5A

To clone the genes of hepatocyte protein interacting with hepatitis C virus NS3 protein, "bait" plasmids of hepatitis C virus NS3 were constructed. After verifying that hepatitis C virus NS3 protein could be steadily expressed in AH109 yeast strain, yeast two hybrid assay was berformed by mating AH109 with Y187 that pre transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X ? gal activity. Nineteen yeast colonies that could grow on QDO and had ? gal activity were obtained, then the library plasmids were extracted and sequenced. The gene sequences from the 19 positive colonies were aligned with the genes deposited in GenBank. It was found hepatitis C virus NS3 protein could interact with some proteins which have different functions.

Interleukin 18 (IL 18) is emerging as a powerful, pleiotropic cytokine involved in determining the polarization of T cell responses. To identify the effect of IL 18 on hepatitis B virus, mouse IL 18 gene was transferred and expressed in 2.2.15 cells. Meanwhile, the inhibition of HBsAg, HBeAg and HBV DNA was investigated. Reverse transcription polymerase chain reaction (RT PCR) was used to amplify the mouse IL 18 from spleen cell of Balb/c mouse challenged with lipopolysaccharide (LPS) and phytahematoagglutinin (PHA), and reconstruct plasmid pLXSN IL18 with the retroviral vector pLXSN. Both pLXSN IL18 and pLXSN were transfected into PA317 cells and then 2.2.15 cells were infected by using the supernatant containing pseudovirus released by PA317. HBsAg and HBeAg were detected by ELISA and the HBV DNA was determined by quantitative PCR. The 580bp of mIL 18 was cloned into retroviral vector pLXSN. The pseudovirus contained in the supernatant of transfected PA317 cells was identified by RT PCR. When the pseudovirus was infected into 2.2.15 cells and incubated for 3, 5, 7, 14 days, the P/N value of HBsAg and HBeAg decreased gradually and became negative on day 14. The copies of HBV DNA reduced markedly. The results indicate that mIL 18 gene expressed in 2.2.15 cells can significantly inhibit the replication expression of HBV and may be used as a potential agent for gene therapy against HBV infection.

Currently, the resistance of first-line anti-tuberculosis drugs has made the prevention and treatment of tuberculosis increasingly difficult, posing a serious threat to global public health. Several studies have shown that efflux pumps are one of the important causes for bacteria to develop multi-drug resistance and extremely-drug resistance, and efflux pump inhibitors can inhibit the efflux of antibacterial drugs, thereby reducing bacterial drug resistance. Numerous natural products and synthetic compounds have been reported to possess efflux pump inhibitory activity, but they have not been applied in clinical settings because of their toxicity, pharmacokinetic properties, etc. Therefore, we summarized the efflux pump inhibitory activity, antimicrobial activity, and structure-activity relationships of reported efflux pump inhibitors against Mycobacterium tuberculosis in recent years, providing references for the development of new efflux pump inhibitors with better activity and lower toxicity.

Objective To study the relationship of tumor necrosis factor beta (TNF-?) polymorphism with human IgA nephropathy of Chinese Han nationality in Inner Mongolia Autonomous Region. Methods The A→G single base mutation polymorphism in TNF-?1096 locus were analyzed among 80 normal controls and 79 IgA nephropathy patients by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The genotype frequencies of TNF?* 1/1 and TNF?* 2/2 in IgA nephropathy patients were significantly higher than that in normal controls (x1/12=5.58,P

Objective The purpose of the present study is to illustrate the effect of olfactory ensheathing cells(OECs) on the survival and neurite outgrowth of GABAergic neurons in vitro. Methods OECs were dissociated from olfactory bulb and neurons from spinal cord of E12 mouse. On the sixth day in vitro,the Millipore cultue blank with OECs was transferred to the neuron culture mediam and continue the co-culture for another 6 days.The cultured neurons were stained with anti-GABA antibody.The neurite of neurons was observed with an image system.The number of GABAergic positive neurons was counted under the microscope. Result The number of GABAergic neurons was 39^7?6^3 in co-culture groups,whereas the number of GABAergic neurons represented only 27^6?2^7 in control groups(CG),(P

Abstract： Objective　To analyze the species distribution of microorganisms in culture tubes reported positive byBACTEC™ MGIT960 (hereafter referred to as "MGIT960") after species identification using matrix-assisted laser desorption/ionization time of flight massspectrometry (MALDI-TOF MS) technique. Methods　From 2021 to 2022, a total of 2 662 positive tubes reported by the MGIT 960 instrument at Tuberculosis Reference Laboratory of Tianjin Center for Tuberculosis Control were collected. Liquid cultures were independently inoculated to blood plate and neutral L-J medium, and the resulting isolate strains were identified using the MALDI-TOF MS method. According to the MALDI-TOF MS results, the non-repetitive results of the same patient on the same culture medium were analyzed for the composition ratio of strain distribution. For the strains not identified by MALDI-TOF MS, 38 strains were selected for 16S rRNA gene sequencing. Results　A total of 605 isolates were obtained from blood plates, and 501 of those were analyzed. Among them, Mycobacterium accounted for 17.76% (89/501), predominant by Mycobacterium abscess 10.18% (51/501) and Mycobacterium fortuitum 3.19% (16/501). Bacteria other than Mycobacterium accounted for 68.06% (341/501), with the main ones being Nocardia farcinica 15.57% (78/501), Gordonia sputa 9.38% (47/501) and Gordonia bronchialis 7.58% (38/501). There were 71 unidentifiable strains, making up 14.17% (71/501). A total of 2 378 strains were isolated from neutral L-J mediums, 1 748 of which were used in the incoming analysis. Among these, 78.72% (1 376/1 748) were Mycobacterium, 60.53% (1 058/1 748) were Mycobacterium tuberculosis (MTB), 4.69% (82/1 748) were Mycobacterium chimaer intracellulare group and 3.55% (62/1 748) were Mycobacterium Lentiflavum. Bacteria other than Mycobacterium accounted for 17.11% (299/1 748) in neutral L-J medium isolates, with Nocardia farcinica 4.35% (76/1 748), Gordonia sputa 2.69% (47/1 748) and Gordonia bronchialis 2.12% (37/1 748) as the main species. There were 73 strains that couldn't be identified, comprising 4.18% (73/1 748). The 38 strains that not identified by MALDI-TOF MS were all found to be Actinobacteria and Firmicutes by sequencing. Conclusions　A variety of nontuberculous Mycobacterium and bacteria other than Mycobacterium were found in the positive culture tubes reported by the MGIT960 instrument, most of which could be quickly identified by mass spectrometry. Bacteria other than Mycobacterium are mainly Nocardia and Gordonia, which should be paid attention to in differential diagnosis.

Objective:To establish a lung cancer model of patient-derived tumor xenografts (PDTX) and to explore the relation-ship between primary cisplatin resistance and ERCC1 and IGFBP5 expression levels. Methods:Lung cancer tissues from 84 patients who underwent surgery were collected and implanted into nude mice. Patient characteristics for the first generation xenografts that were and were not engrafted were compared. Passage 3 xenografts were treated with cisplatin. The expression levels of ERCC1 and IGFBP5 in cisplatin-resistant and cisplatin-sensitive groups were detected using immunohistochemistry assay. Results:The model success rates were 32.14%(27/84) in first-generation xenografts, 88.89%(24/27) in second-generation xenografts, and 95.83%(23/24) in third-gener-ation xenografts. The tumorigenicity of first-generation xenografts was correlated with size, differentiation, clinical stage, and histologi-cal type. PDTX tumors maintain the histological type of parental tumors through serial passage in nude mice. ERCC1 expression level was significantly higher in the cisplatin-resistant group than in the cisplatin-sensitive group, whereas the IGFBP5 expression level was lower in the cisplatin-resistant group than in the cisplatin-sensitive group. Conclusion:Lung cancer PDTX models were successfully es-tablished, and histological characteristics of the primary cancers were retained. Therefore, the models may serve a function in preclini-cal research of lung tumor biology and for exploring the drug resistance mechanism of tumors. The cisplatin resistance of primary lung cancer may be correlated with the expression level of ERCC1 and IGFBP5 in lung carcinoma.

OBJECTIVE:To prepare the vinorelbine-tetrandrine liposomes modified with RGD,and study the inhibitory effect on glioma C6 cells. METHODS:Film dispersion method and ammonium sulfate gradient method were used to prepare the vinorel-bine-tetrandrine liposomes modified with RGD,and the morphology and particle size distribution were observed. The vinorelbine content was determined,and sulforhodamine B method was used to respectively determine the inhibitory effects of blank targeting liposomes,normal vinorelbine liposomes and vinorelbine-tetrandrine liposomes modified with RGD on C6 cells. RESULTS:The prepared vinorelbine-tetrandrine liposomes modified with RGD were spherical or almost spherical with smooth surface,and particle size was about 120 nm. The average content of vinorelbine was 28.27 μg/mL(RSD=0.38％,n=3). Blank targeting liposomes had no significant effect on the growth of C6 cells;vinorelbine-tetrandrine liposomes modified with RGD can obviously inhibit the growth of C6 cells,and cell viability after its effect was significantly lower than normal vinorelbine liposomes (P<0.05). CON-CLUSIONS:Vinorelbine-tetrandrine liposomes modified with RGD are successfully prepared,and they show obvious inhibitory ef-fects on the growth of C6 cells.

Objective To realize rapid and non-destructive drug classification and improve the accuracy of drug classification.Methods A model for drug classification based on the combination of principal components analysis and artificial neural network (PCA-ANN) method was introduced.The software for drugs classification was then developed with the utility of MATLAB language.The near infra-red spectrum (NIRS) detection technique was executed on five kinds of drugs (a total of 120 batch samples) and the detection data was collected within the range of 1 350-1 800 nm of excitation wavelength and 0.5 nm of wavelength interval.Results The network training mean square error (MSE) was 5.91e-03,and the prediction error (β) was 2.469％ when the number of the interfering drugs number was less than 5.Conclusions The classification of drugs by NIRS combined with PCA-ANN is feasible and the classification accuracy can be increased.