Abstract: Objective　To investigate the diagnostic efficacy of rifampin-resistant real-time fluorescent quantitative nucleic acid amplification detection technology (GeneXpert MTB/RIF) in bronchoalveolar lavage fluid (BALF) combined with peripheral blood tuberculosis infection T cell spot test (T-SPOT) and tuberculosis antibody (TB-Ab) in smear-negative pulmonary tuberculosis. Methods　The clinical data of 114 cases of clinically diagnosed smear-negative pulmonary tuberculosis, 80 cases of non-tuberculous pulmonary diseases and 22 cases of smear-positive pulmonary tuberculosis in our hospital from January 2019 to January 2021 were retrospectively analyzed. The detection results of peripheral blood T-SPOT, TB-Ab and BALF GeneXpert in the three groups were analyzed. The sensitivity, specificity, negative predictive value, positive predictive value, false negative rate, false positive rate and Youden index of the three detection methods were compared. The differences in the positive detection rate of smear-negative pulmonary tuberculosis between the separate detection and the combined detection of the three methods were compared. The receiver operating characteristic curve (ROC) was performed to calculate the area under the curve (AUC). Results　The sensitivity of BALF GeneXpert and peripheral blood T-SPOT and TB-Ab was 66.91%, 80.88% and 90.44%, respectively. The specificity was 98.75%, 73.75% and 41.25%, respectively; the diagnostic coincidence rates were 78.70%, 78.24% and 72.22%, respectively, which were higher than 70.00%. In the smear-negative pulmonary tuberculosis group, the positive detection rates of these three methods in the smear-negative pulmonary tuberculosis group were 63.15%, 79.82% and 90.35%, respectively, and the differences were statistically significant compared with those in the non-tuberculosis pulmonary disease group (all P<0.01). The positive detection rate of the three combined methods in the smear-negative pulmonary tuberculosis group was 96.49 %, which was significantly higher than that of TB-GeneXpert method and T-SPOT, and the differences were statistically significant (χ2=37.283, P<0.01; χ2=13.612, P<0.01); the Youden index of combined detection was significantly higher than that of single detection, and the AUC of combined detection was 0.977, which was significantly higher than that of single detection. Conclusion　BALF GeneXpert combined with peripheral blood T-SPOT and TB-Ab can significantly improve the diagnostic rate of bacterial-negative pulmonary tuberculosis, providing a strong basis for guiding clinical treatment.

Objective To study the pulmonary protective effect of large dose of ambroxol hydrochloride injection in postoperative patients with severe hypertensive intracerebral hemorrhage.Methods One hundred and sixty postoperative patients with hypertensive intracerebral hemorrhage were selected,and the patients were divided into conventional dose group and large dose group by random digits table method with 80 cases each.The therapeutic effect,partial pressure of oxygen,oxygenation index,acute physiology and chronic health evaluation Ⅱ score,antibiotic application time and hospitalization period in intensive care unit were compared between 2 groups.Results After treatment,the partial pressure of oxygen and oxygenation index in large dose group were significantly better than those in conventional dose group:(94.7 ± 7.2) mmHg (1 mmHg =0.133 kPa) vs.(86.5 ± 8.1) mmHg and (420.3 ± 35.5) mmHg vs.(356.0 ± 28.7) mmHg,the acute physiology and chronic health evaluation Ⅱ score was significantly lower than that in conventional dose group:(8.2 ± 3.4) scores vs.(11.6 ± 3.5) scores,the antibiotic application time and hospitalization period in intensive care unit were significantly shorter than those in conventional dose group:(8.5 ± 3.7) d vs.(13.4 ± 5.8) d and (7.3 ± 2.5) d vs.(9.7 ± 5.2) d,and there were statistical differences (P＜ 0.05).The total effective rate in large dose group was significantly higher than that in conventional dose group:92.5％ (74/80) vs.83.8％ (67/80),and there was statistical difference (P ＜ 0.05).Conclusion The pulmonary protective effect of large dose of ambroxol hydrochloride injection is significant in postoperative patients with hypertensive intracerebral hemorrhage.

OBJECTIVETo study the expression and switching of Th1/Th2 cytokines gene in human gliomas and its effects on occurring and developing of human gliomas.

METHODSInterleukin (IL)-2 and interferon-gamma represent Th1 type cytokines. IL-4, IL-6, IL-10, and IL-13 represent Th2 type cytokines. The gene expressions of Thl/Th2 cytokines in human glioma cells, glioma infiltrating lymphocytes, and glioma cell lines were detected by reverse transcription polymerase chain reaction (RT-PCR). The biological activity of cytokines in the supernatant of glioma cell lines was assayed by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe total positive rates of Th1 and Th2 type cytokines gene in human glioma cells were 14.77% and 75%. The total positive rates of Th1 and Th2 type cytokines gene in glioma infiltrating lymphocytes were 22.73% and 68.17%. There was obviously predominant expression of Th2 type cytokines in human glioma tissues, glioma infiltrating lymphocytes, and glioma cell lines. There was no unbalanced expression of Th1/Th2 cytokines in normal brain tissues.

CONCLUSIONThere is a predominant expression of Th2 type cytokines in human glioma cells. The switching of Thl/Th2 cytokines gene may play an important role in the occurring and developing of human gliomas.

Adolescent ; Adult ; Aged ; Astrocytoma ; metabolism ; pathology ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Child ; Child, Preschool ; Cytokines ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Glioblastoma ; metabolism ; pathology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Male ; Middle Aged ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism

The characteristics of On-line Near Infrared Spectroscopy Technology are introduced and its applications in various industries, such as petro-chemical, pharmaceutical, agriculture and food are reviewed. Considering the features in TCM manufacturing, the application potentials of On-line Near Infrared Spectroscopy Technology in Manufacturing TCM are forecasted and the key problems to be solved are discussed.
Drugs, Chinese Herbal ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Technology, Pharmaceutical ; methods

Objective To study the role of arrestin domain-containing protein 4( ARRDC4) in regulation of enterovirus 71(EV71) triggered innate IL-6 production and the underlying mechanism .Methods THP-1-derived macrophages (t-M?) were transfected with ARRDC4 specific siRNA and negative control siRNA .The expression and production of IL-6, replication and virus titer of EV71, and the activation of signaling pathway adaptors were analyzed with quantitative real -time PCR, ELISA and Western blot.Results Upon EV71 infection, ARRDC4 was upregulated.ARRDC4 silencing could enhance mRNA expression and production of IL-6, thus increasing the replication and virus titer of EV71.In ARRDC4 silenced t-M?, the activation of p-65,IκBα,ERK,JNK and p38 was promoted.Conclusion ARRDC4 promotes EV71-t riggered IL-6 production by enhancing the activation of NF-κB and MAPK signaling pathway to inhibit EV71 infection, contributing to positive regulation of anti-EV71 innate immune responses .

Objective To analyze the effect of video-assisted thoracoscopic surgery and traditional surgery for multiple rib fractures. Methods 102 cases of patients with multiple rib fractures from January 2014 - December 2017 was selected and divided into two groups after different therapy: 51cases in control group were treated with traditional surgery, 51cases in study group were treated with thoracoscopic assisted surgery, the clinical results was compared and analyzed. Results The observation group of intraoperative bleeding, postoperative drainage were significantly higher than the control group (P < 0.05); the operation time of observation group was significantly longer than the control group, with significant differences between the two groups (P < 0.05); the study group of pain time (8.39 ± 2.24) d, ambulation time (5.76 ± 2.43) d and the hospitalization time (10.53 ± 3.51) d was significantly shorter than the control group, the difference between the two groups was statistically significant (P < 0.05), study group complication rate was 3.92% and the control group complication rate was 17.64% with significant difference (P < 0.05). Conclusion The clinical effect of video-assisted thoracoscopic surgery in patients with multiple rib fractures is significant, can prevent complications, reduce medical costs, it is worth learning.

Objective To prepare T-helper cell(Th)epitnpe-modified human soluble B cell activating factor belonging to the tumor necrosis factor family(BAFF)mutants and evaluate their immune response in vaccinated mice.Methods Recombinant cDNAs were cloned and ligated into the prokaryotie expression vec- tor pQE-80L respectively.The recombinant proteins were induced to express by IPTG in E.coli DH5?and purified with Ni-NTA chromatography.BALB/c mice were immunized with recombinant proteins respectively and the titres of the antibodies that were cross-reactive with BAFF in sera were analyzed by ELISA.Inhibiting ability of the antibodies in sera was analyzed by MTT assay.Results The recombinant proteins were highly expressed in E.coli DH5?.After purification,the purity of recombinant proteins was more than 90%.BALB/c mice immunized with recombinant proteins produced high levels of BAFF-specific antibodies.Cell proliferation assay showed that the sera of immunized mice could inhibit the proliferation-inducing activity of recombinant sBAFF and natural sBAFF.Conclusion The immune inhibitors of human BAFF which can induce polyclonal antibodies that are cross-reactive with BAFF are successfully prepared.These results may provide the basis for further study of their therapeutic effects.

This study was aimed to construct mouse Fas-targeting si RNA-expressing recombinant retroviral vector in order to explore the therapeutic potential of Fas inhibition by siRNA in the treatment of aplastic anemia and to provide a basia for extensive development of RNA interference techninque. The U6(+) 27 promoter cassette and siFas sequence were obtained by PCR method. The U6-siFas fragment was cloned into the multiple restriction site of pLXSN-EGFP and directly downstream of EGFP gene. The resultant retroviral vector pLXSN/EGFP-siFas was packaged using PA317 cell line and tittered using NIH3T3 cell line. P815 cells were infected by the retroviral vector. EGFP expression in P815 was observed under fluorescent microscope and Fas inhibition effect was detected by immunohistochemistry. The results indicated that successfully constructed retrovirus vector pLXSN/EGFP-siFas was could not only deliver siRNA into mammalian cells efficiently and inhibit Fas expression in P815 cells, but also could express EGFP as marker and neomycine resistance gene to allow antibiotic selection. It is concluded that the successful construction of this retroviral vector would greatly facilitate the application of RNA interference and lay the foundation for therapeutic study of Fas inhibition in the treatment of aplastic anemia.
Anemia, Aplastic ; genetics ; pathology ; therapy ; Animals ; Cell Line ; Cell Line, Tumor ; Cloning, Molecular ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Immunohistochemistry ; Mice ; Microscopy, Fluorescence ; NIH 3T3 Cells ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Retroviridae ; genetics ; Transfection ; fas Receptor ; genetics ; metabolism

Combined hepatocellular-cholangiocarcinoma (CHC) is a mixed tumor containing elements of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC).Its remarkable histological heterogeneity has been linked to putative hepatic progenitor cell (HPC) origin.However,detailed histological or phenotypic description is rarely documented.In the present study,we reassessed 68 cases previously diagnosed as hepatitis B-related CHCs by immunohistochemistry and double-fluorescence immunostaining,focusing on HPC associated phenotypic observation of intermediate area of the tumor.It was found that tumor cells showed remarkable heterogeneity in intermediate area.Tumor cells with intermediate morphology between hepatocytes and cholangiocytes were oval-shaped and small with scant cytoplasm and hyperchromatic nuclei,arranging in solid nests mostly.By Keratin 7 (K7) staining,it appeared that the nests of tumor cells represented a maturation process from the undifferentiated small cells to mature hepatocytes through the "transitional" cells.Then,these small cells were further confirmed with intermediate phenotype as HPC by exploring immature hepatocellular marker and HPC/biliary markers co-localization.In conclusion,the HPC associated trait in CHC can be interpreted by HPC origin or gain of"stemness" by dedifferentiation.It is still too soon to give a final word that it is innate or acquired signature of HPC associated trait in CHC.

Objective To determine the prevalence and genotype of hepatitis E virus (HEV)among commercial swine population in Eastern and Southern China. Methods Six hundred specimens of swine bile collected from 5 slaughterhouses in Eastern and Southern China from 2007 to 2009 were tested for HEV RNA using nested RT-PCR. PCR products were sequenced for phylogenetic analysis. Results Forty-seven out of the 600 samples (7.83%) were positive for HEV RNA. Based on the 150 nt fragment within HEV ORF2, data from phylogenetic analysis revealed that all the 47 HEV isolates were identified to be genotype Ⅳ, sharing 75.0%-83.4%, 75.0%-84.6%, 71.9%-80.7% and 88.1%-91.5% nucleotide identities with prototype Ⅰ,Ⅱ,Ⅲ and Ⅳ HEV strains respectively while majority of the isolates clustered within their respective isolation sites. Conclusion HEV was widespread in commercial swine population in Eastern and Southern China that raised a serious concern about the safety regarding the consumption of pork products.