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Yi-Ran Han,1,* Tian-Hao Wang,2,* Wen-Ping Gong,3,* Jian-Qin Liang,1 Hui-Ru An1 1Department of Tuberculosis, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, Beijing, 100091, People’s Republic of China; 2Department of Emergency, The Eighth Medical Center of PLA General Hospital, Beijing, 100091, People’s Republic of China; 3Tuberculosis Prevention and Control Key Laboratory, Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute for Tuberculosis Research, Senior Department of Tuberculosis, The 8th Medical Center of Chinese PLA General Hospital in China, Beijing, 100091, People’s Republic of China*These authors contributed equally to this workCorrespondence: Hui-Ru An; Jian-Qin Liang, Department of Tuberculosis, Senior Department of Tuberculosis, The Eighth Medical Center of PLA General Hospital, No. 17 of Heishanhu Road, Haidian District, Beijing, 100091, People’s Republic of China, Tel +86 10 66775098, Email anhuiru25@outlook.com; wliangjianqin@126.comObjective: To make a systematic evaluation of the clinical efficacy of thymopentin combined with antituberculous drugs in treating drug-resistant pulmonary TB (PTB).Methods: Relevant studies were retrieved from PubMed, Embase, Cochrane Library, Chinese Biomedical Literature Database, CNKI, and Wanfang Database. STATA software was used to evaluate the differences in focal absorption rate, the time to cough symptom remission, sputum culture-negative rate, CD3+ T, CD4+ T, and CD8+ T cell levels after treatment.Results: A total of 23 randomized controlled trials literature involving 2031 cases were included. Meta-analysis revealed that compared with conventional therapy, the sputum culture-negative rate was significantly increased after 2– 3 months and 6– 9 months of treatment and the whole course of combined thymopentin treatment. The risk ratio (RR, 95% CI) was 1.44 (1.26– 1.64), 1.47 (1.21– 1.78), and 1.27 (1.18– 1.36), respectively. In the combined thymopentin treatment group, the focal absorption rate was higher, with RR (95% CI) = 1.36 (1.25– 1.47), the time of cough remission was shortened, with WMD (95% CI) =− 9.46d (− 10.36,− 8.57) and the differences were all statistically significant. Combined thymopentin therapy could effectively improve the levels of CD3+ T and CD4+ T lymphocytes in patients with drug-resistant PTB after 2– 3 months, 6– 9 months of treatment. The WMD (95% CI) were 9.96% (7.84, 12.08), 4.68% (2.90, 6.47) and 10.26% (7.81, 12.71), 7.21% (6.28, 8.15), respectively, and could also reduce the level of CD8+ T lymphocytes after 2– 3 months and 6– 9 months of treatment. The WMD (95% CI) were − 4.06% (− 4.96, − 3.13), − 3.52%, (− 4.07,− 2.98), respectively, and the differences were all statistically significant.Conclusion: Thymopentin adjuvant treatment for drug-resistant PTB can promote the therapeutic effect and improve the immune indexes in patients with drug-resistant PTB.Keywords: thymopentin, drug resistance, pulmonary TB, immunity, meta-analysis

This paper was to investigate the effect of Huanglian Jiedu Decoction(HLJD) on ulcerative colitis(UC) in mice, and determine the effective components in plasma, and virtually screen its therapeutic target, and predict its mechanism. Sixty Balb/c mice were randomly divided into blank group, model group, mesalazine treatment group(0.3 g·kg~(-1)), and HLJD treatment groups(24.66, 12.33, 6.17 g·kg~(-1)). Excepted for the blank group, all the mice in HLJD and mesalazine treatment groups were gavage administration. All mice freely drank 2.5% DSS solution for seven days to induce UC. The disease activity index(DAI) was detected each day. At the end of the experiment, HE staining was used to observe the pathological changes in colon. The content of IL-1β, IL-6 and TNF-α in colon were determined by ELISA. The effective components in plasma were determined by UPLC-Q-TOF-MS. The reverse docking in PharmMapper was used to screen the component targets. The disease targets of UC were collected by searching TTD, OMIM and GeneCards databases. The intersection of the component targets and disease targets was selected as the therapeutic targets. Then the therapeutic targets were imported into the STRING for GO and KEGG enrichment analysis. Discovery Studio was used to simulate the docking between the components and the targets. RESULTS:: showed that the DAI in the model group increased significantly(Plt;0.05), and the number of inflammatory cells and infiltration degree increased significantly compared with the blank group. The DAI in HLJD treatment group was significantly reduced(Plt;0.05), and the number and infiltration degree of inflammatory cells were reduced compared with the model group. The ELISA results showed that the levels of IL-1β, IL-6 and TNF-α were increased significantly in the model group(Plt;0.01) compared with the blank group, and significantly down regulated in the HLJD treatment group(Plt;0.05) compared with the model group. After UPLC-Q-TOF-MS analyse, ten components were identified. The network pharmacology analysis showed that the action targets were significantly enriched in 129 of biological processes, such as response to organic substance, chemical and oxygen-containing compound, etc., as well as 16 of signal pathways, such as IL-17, TNF and hepatitis B signal pathways, were enriched too. The results of molecular docking showed that limonin, palmatine and berberine could bind to CASP3 and MMP9 by hydrogen bond. In conclusion, HLJD could alleviate the colonic mucosal inflammatory infiltration and mucosal damage in UC mice. The mechanism may be related to the anti-inflammatory effect on UC mice by reducing the levels of IL-1β, IL-6 and TNF-α in colon through limonin, palmatine and berberine regulating IL-17 signal pathway and TNF signal pathway via CASP3 and MMP9 meditated.

This paper was to investigate the effect of total flavonoids of Lichi Semen(TFL) on carbon tetrachloride(CCl_4)-induced liver fibrosis in rats, analyze and predict its mechanism of action and potential quality markers(Q-marker). Firstly, male SD rats were taken and injected subcutaneously with a 40% CCl_4-vegetable oil solution twice a week for 8 consecutive weeks to establish a rat model of liver fibrosis. The rats with liver fibrosis were randomly divided into model group, silybin group(43.19 mg·kg~(-1)), Fuzheng Huayu Capsules group(462.75 mg·kg~(-1)), and TFL groups(100 mg·kg~(-1) and 25 mg·kg~(-1)), with normal rats as a blank group, 10 rats in each group. Except for the blank group, the rats in the other groups were subcutaneously injected with 40% CCl_4-vegetable oil solution of a maintenance dose, once a week. The rats in various treatment groups received corresponding doses of drugs, while the rats in the blank group and model group received the same volume of normal saline once a day for 4 weeks. At the end of the experiment, blood was collected from the abdominal aorta and the liver tissues were collected. The levels of total bilirubin(TBiL), direct bilirubin(DBiL), indirect bilirubin(IBiL), alanine aminotransferase(ALT), and aspartate aminotransferase(AST) in serum were detected by using an automatic biochemical detector. Masson staining was used to observe the histopathological changes of rat liver. Then, the chemical compositions of TFL were collected, and the action targets of these chemical compositions were predicted through SWISS database and reverse molecular docking server(DRAR-CPI). After screening of disease targets of liver fibrosis by Gene Cards database, the protein-protein interaction was analyzed with use of STRING database, and GO(gene ontology) analysis and KEGG(Kyoto encyclopedia of genes and genomes) enrich analysis were also carried out. Moreover, an iTRAQ proteomics technology was used to determine protein expression in liver tissues of rats in TFL, model and blank groups to verify the targets. Furthermore, Cytoscape software was used to establish and visualize the network of chemical components, targets and pathways, and predict the potential Q-marker of TFL. The results showed that the levels of TBiL, DBiL, IBiL, ALT, and AST in the model group were significantly higher than those in the blank normal group(Plt;0.05), and the above levels in the treatment groups were lower than those in the model group, but with no significant differences. Masson staining showed that the liver damage and the degree of fibrosis were severe in the model group, and were relieved to different degrees in the treatment groups. Then, 74 chemical components were screened, which could act on 865 targets such as EGFR and SRC, participating in the regulation of cancer pathways, PI3 K-Akt signaling pathway, HIF-1 signaling pathway and other signaling pathways closely related to liver fibrosis. Pinocembrin, quercetin, epicatechin, procyanidin A2, naringenin, nobiletin, phlorizin and rutin showed the highest correlation with liver fibrosis-related targets and pathways. Proteomics results showed that a total of 18 proteins among the 45 proteins predicted by internet pharmacology were identified, among which 6 proteins were significantly expressed, including 5 up-regulated proteins and 1 down-regulated protein. The protein expression of ALB, PLG, HSP90 AA1, EGFR and MAP2 K1 was significantly returned to a normal state in the TFL treatment groups. In conclusion, TFL may demonstrate the anti-hepatic fibrosis and potential hepatoprotective effects by regulating the expression of ALB, PLG, HSP90 AA1, EGFR and MAP2 K1, which may be associated with the regulation of multiple signaling pathways related to liver fibrosis such as PI3 K-Akt pathway. Pinocembrin, quercetin, epicatechin, procyanidin A2, naringenin, nobiletin, phlorizin and rutin could be regarded as potential Q-markers of TFL for quality control.

Background This research aimed to investigate the clinical value and characteristics of peripheral blood lymphocyte subsets in tuberculosis (TB) patients by flow cytometry. Methods The Absolute counts of T, CD4+ T, CD8+ T, NK, NKT and B lymphocytes were detected in 217 cases of pulmonary TB (PTB), 163 cases of PTB & extra-PTB and 65 cases of extra-PTB patients. We analyzed the change characteristic of subset counts with the clinical parameters in PTB and compared the subsets differences among PTB, PTB with extra-PTB, and extra-PTB patients. Results The absolute counts of six subsets in 75.3% of PTB patients were lower than the normal reference range, and 44% patients showed lower CD4+ lymphocytes. (1) The absolute counts of T, CD4+ T, CD8+ T and B lymphocytes was significantly lower in patients older than 60 years old. (2) The NKT cell counts was significantly lower in female patients than the males; (3) 40.8% of patients with positive etiological results and 49.2% of extra-PTB patients showed lower CD8+ T and NK cell counts below the reference range respectively. (4) The T and CD4+ T lymphocyte counts in PTB with positive IGRA were significantly higher than those with negative IGRA; (5) The T and CD8+ T cell counts in PTB with positive IgG antibody were decreased; (6) The T, CD4+ T, CD8+ T and B cell counts reduced with the increasing lesion lobe numbers, accelerated ESR, complications with anemia and low serum albumin, and NK cells also decreased in patients with anemia and low serum albumin; and the T, CD8+, and B cells significantly decreased in PTB patients with diabetes, while NKT increased. Conclusions The immune function in most TB patients is impaired and the absolute counts of lymphocyte subsets could be used as the evidence for immune intervention and monitoring the curative effect.

The rapidly increasing number of multidrug-resistant tuberculosis (MDR-TB) cases worldwide underlines the necessity for the rational use of key second-line drugs such as kanamycin. In this study, we determined the prevalence of, and risk factors associated with, kanamycin-resistant tuberculosis (TB) in 309 Hospital, Beijing, China, with the aim of providing information for better case management in order to minimize further development of extensively drug-resistant TB (XDR-TB). Drug susceptibility testing results and clinical data were retrospectively analysed for hospitalized TB patients for whom such data were available in 309 Hospital for the period 1997-2009. Univariate and multivariate analyses were used to determine the risk factors associated with kanamycin-resistant TB. During 1997-2009, 553 (14.4 %) of 3843 tested Mycobacterium tuberculosis isolates from hospitalized TB patients were kanamycin-resistant. The increasing trend of resistance to kanamycin was reversed since 2000. The independent risk factors associated with kanamycin-resistant TB included living in urban areas [adjusted odds ratio (OR) = 1.89], being retreated for repeat cases (adjusted OR = 1.60), being smear-positive for acid-fast bacilli at admission to the hospital (adjusted OR = 1.39), having ofloxacin-resistant (adjusted OR = 1.61) or para-aminosalicylic acid-resistant TB (adjusted OR = 1.47), having MDR-TB (adjusted OR = 5.10), having MDR-TB plus ofloxacin resistance (adjusted OR = 4.27) and having poly-resistant TB (adjusted OR = 3.94). The remaining rate of kanamycin resistance is still high despite the reversal of the increasing trend during the past decade. Surveillance of kanamycin resistance, especially among high-risk populations, should be continued to closely monitor trends so that appropriate action can be taken.

To evaluate the relationship between mutations in rpsL or rrs genes and streptomycin (SM) resistance, we compared four molecular methods for their clinical value in the detection of SM resistance. Genotypic analysis of SM resistance in 167 M. tuberculosis clinical strains isolated from Chinese patients was performed by direct DNA sequencing, SSCP, RFLP, and reverse dot-blot hybridization (RDBH) assays. Of the 98 SM-resistant isolates, 78 (79.6%) had missense mutations in codon 43 or 88 of rpsL resulting in a Lys to Arg substitution, 6 (6.1%) had mutations of the rrs gene at positions 513 A to C or T or 516 C to T, and 14 (14.3%) had the wild-type sequence. None of the 69 SM-susceptible isolates examined had alterations in rpsL or rrs. The results of the SSCP, RFLP, and RDBH analyses for these mutations and wild-type sequences were completely consistent with DNA sequencing data. Five distinct single-nucleotide substitutions in codon 43 or 88 of rpsL gene or in position 513 or 516 of rrs gene were correctly identified in 84 of 98 (85.7%) phenotypically SM-resistant isolates by RDBH assay. Molecular analyses of the rpsL and rrs genes are useful for rapid prediction of SM resistance in most clinical strains of M. tuberculosis. Reverse dot-blot hybridization assay is a rapid, simple, and reliable method for the detection of drug resistance.

To optimize preparation process of liuwei cataplasm, a Zhuang folk medical herb popularly applied as a kind of rheumatism, rheumatoid drug in Guangxi Zhuang Autonomous Region.Orthogonal design test was employed to optimise matrix proportion and molding technology of liuwei cataplasma with glossiness, adhesive power, matrix residue, skin following and repeatedly exposing as indexes.Optimum matrix ratio of PANA, pressure sensitive adhesive, CNC-Na, gelatin, dihydroxyaluminium aminoacetate and glycerol was 0.5:1.0:0.4:0.1:0.3:3.0. Optimized parameters of technology were as follows: stiring speed 600 r/min, refining temperature 60 degrees C and drug loading of Liuwei extract 13.04%.Optimized liuwei cataplasm has a good adhesive power, exipcients and glossiness, meanwhile drug loading is large and no skin residue. The process is simple, stable and viable.

To investigate the pharmacokinetics and bioavailability of ginsenoside Rg1 in rats.Ginsenoside Rg1 was oral administered or intravenous administered to each rat. The plasma concentration of ginsenoside Rg1 was estimated by RP-HPLC. 3P97 software was used to calculate pharmacokinetic parameters.Main parameters of ginsenoside Rg1 after oral or intravenous administered were: AUC(0 --t), 322.70 +/- 20.78, 99.76 +/- 8.91 microg x h/mL, CL 0.08 +/- 0.02, 3.01 +/- 0.69 L/(kg x h), V 0.23 +/- 0.01, 22.75 +/- 2.09 L/kg, t1/2alpha 0.48 +/- 0.18, 0.87 +/- 0.21 h, t1/2beta 19.57 +/- 2.81, 18.68 +/- 2.74 h, MRT6.91 +/- 0.99, 8.15 +/- 1.05 h(-1), respectively. The relative oral bioavailability of ginsenoside Rg1 was 2.5%.The oral bioavailability of ginsenoside Rg1 is very low.

To develop a new method, gene array, which can be used for rapid detection of rpoB mutations in Mycobacterium tuberculosis.Probes were designed according to the sequence of Mycobacterium tuberculosis rpoB gene and the gene array was developed. The DNA fragment which contained hot mutation sites of rpoB gene was amplified with biotin-labelled primers by PCR, and then hybridized with gene array. Mycobacterium tuberculosis strain H(37)Rv DNA was used as the control. The rpoB genes in Mycobacterium tuberculosis clinical isolates were also analyzed by polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and PCR-DNA sequencing.We analyzed the rpoB genes of 111 Mycobacterium tuberculosis clinical isolates by PCR-SSCP. Of 70 rifampicin-resistant Mycobacterium tuberculosis isolates, 63 isolates had different SSCP profiles from that of the standard strain H(37)Rv. No difference from the standard strain was found in 41 rifampicin-susceptible and 7 rifampicin-resistant isolates. We also analyzed their rpoB genes by gene array. Of 111 Mycobacterium tuberculosis clinical isolates, the results of gene array in 41 drug-sensitive strains were similar to that in Mycobacterium tuberculosis H(37)Rv. 90% (63/70) rifampicin-resistant strains had rpoB gene mutation. 53% (37/70) rifampicin-resistant strains had serine substitution at codon 531. 21% (15/70) strains had histidine substitution at codon 526. 16% (11/70) strains had amino acids substitution in other position. The results of gene array corresponded with that of PCR-SSCP and DNA sequencing.Gene array might become a rapid, simple, and accurate method for detecting rpoB mutations in most of the rifampicin-resistant Mycobacterium tuberculosis.