Objective　To explore the molecular epidemiological characteristics of local Mycobacterium tuberculosis in Chongqing, China using genotyping methods and establish a suitable genotyping system tuberculosis genotyping in the region. Methods A total of 268 isolates collected from the sputum of tuberculosis patients at two national drug resistance monitoring sites in Chongqing from 2019 to 2021. The DNA of these isolates was extracted, and genotyping of the isolates was performed using VTNR 24 locus typing method. Epidemiological investigations were conducted on clustered isolates, and the clustering rates of isolates from the two surveillance sites were compared to analyze their transmission differences. Results In the two regions, the Hunter-Gaston discriminatory index (HGI) in Fengjie County ranged from 0 to 0.85, with 10 loci having an HGI above 0.6, 11 loci with an HGI between 0.3 and 0.6, and 3 loci with an HGI below 0.3; the highest resolution site was MIRU31, while the lowest resolution site was MIRU24. The HGI index in Fengdu County ranged from 0 to 0.81, with 12 loci having an HGI above 0.6, 9 loci with an HGI between 0.3 and 0.6, and 3 loci with an HGI below 0.3; the highest resolution site was MIRU26, while the lowest resolution site was MIRU23. In the clustering analysis, among the 140 isolates from Fengdu County, 10 samples clustered into 4 clusters, with the largest cluster consisting of 4 samples, resulting in a clustering rate of 4.1%. Among the 128 isolates in Fengjie County, 10 samples clustered into 5 clusters, with a clustering rate of 4.2%. Conclusion In Fengdu County and Fengjie County of Chongqing, the main population of the disease show independent genotypes, indicating a low recent transmission rate. The incidence of patients is mainly endogenous recurrence, that is, latent infection develops into active tuberculosis. While conducting standardized management of tuberculosis patients, prevention and control agencies should also increase the screening of latent infection at two monitoring points and carry out preventive treatment of latent infection, which is the key to reduce the incidence of tuberculosis in the two places.

The seeds of Rabdosia rubescens were as the materials to research the impacts of different lead (Pb2+) concentrations(0, 135, 270, 540, 1 080 mg x L(-1)) on seed germination and seedling growth. The results show that: Low concentration of lead had no obvious effect on early germination of the seed, the germination vigor and germination speed were lightly higher but not significantly differed at the level of Pb concentration 135 mg x L(-1) with control group; Mid-high concentration of Pb solution (270-1 080 mg x L(-1)) significantly inhibited the seed germination and seedling growth, which reduced the seed germination rate, germination vigor, germination index, embryo root length and shoot length, growth index with increasing of Pb concentrations. There was a inhibitory effect on embryo shoot length and root length at mid-high lead concentrations stress, and stronger inhibitory effect on root , which was more sensitive than shoot to Pb stress(P < 0.05). Pb bioaccumulation coefficient (BC) was 0.76-2.59, increased with concentration of Pb; Pb enrichment in seedling mainly caused the growth inhibition. The fitting model predictive analyses show, the critical concentration of Pb, which causes the germination rate and biomass fresh weight reducing 10%, is 195.18, 101.65 mg x L(-1).
Germination ; drug effects ; Isodon ; drug effects ; growth & development ; Lead ; toxicity ; Seedlings ; growth & development ; Seeds ; growth & development ; Stress, Physiological

AlM: To silent hypoxia inducible factor-1α ( HlF-1α) gene in malignant melanoma of the choroid cell by small interference RNA ( siRNA ) and investigate its effect on the expression of matrix metalloproteinase-2 ( MMP-2 ) in the choroid cell line human uveal melanoma cell (OCM-1) in hypoxia environment.METHODS:OCM-1 cells cultured on culture flask were divided into normal group and hypoxia group. Hypoxia group were divided into five groups: simple hypoxic group, and interference group, and negative control group, and positive control group, and liposome group. Normal group cells were cultured on DMEM culture flask with 10% FBS, 100U/mL penicillin, 100μg/mL streptomycin as well as high concentration of glucose. The cells were maintained at 37℃ in a humidified 5% CO2 incubator. Cells in good condition were selected for experiment. For hypoxia group, chemical hypoxia inducer CoCl2 was added into nutrient medium at the concentration of 100μmol/L to simulate hypoxia microenvironment. We designed and synthesised siRNA ( siRNA + negative control+positive control ) , the target sequences of the HlF-1α to transfect hypoxic malignant melanoma of the choroid cell. SiRNA including HlF-1α siRNA, β-actin siRNA and negative control group synthesized in vitro transfected hypoxic OCM - 1 cell through Lipofectamine2000. The expression of HlF-1α, MMP-2 gene and the protein were detected by RT-PCR and Western blot. RESULTS: Compared with the normal group, the expression of HlF-1α mRNA was not obviously changed (P>0. 05), but the expression of HlF-1α protein and MMP- 2 mRNA protein was significantly higher ( P<0. 05) . Compared with the other hypoxia groups,β-actin mRNA expression of positive control group decreased ( P< 0. 05 ) , which proved successful transfection. The expression of HlF-1α mRNA and the expression of its protein and both MMP-2 mRNA and its protein was significantly lower ( P < 0. 05 ). The negative control group, liposome control group had no significant difference in the detection of factors (P>0. 05).CONCLUSlON: Hypoxia status may upregulate the HlF-1α in OCM-1 cells by increasing the expression of protein. Hypoxia can also inactivate MMP-2, resulting in upregulation of MMP-2 RNA and the expression of its protein. The expression of HlF-1α and MMP-2 mRNA can be down-upregulated by transfecting OCM-1 with HlF-1α siRNA.

This study was aimed to investigate the T cell (helper T cells) immune status in ITP patients and its relation with therapeutic response. 20 de novo ITP patients were enrolled (8 males, 12 females) with a median age of 41 (20 to 81). Real-time RT-PCR method was used to measure the gene expression of Th cells including T-bet, IFN-γ, GATA-3, TGF-β, Foxp3, IL-2, IL-4 in PBMNC of ITP patients before and after conventional dose of prednisone therapy [1 mg/(kg·d)] and in PBMNC of 20 normal controls. The results showed that T-bet, IFN-γ and IL-2 were significantly over-expressed in PBMNC of ITP patients before treatment compared with that in normal controls (p < 0.01), and compared with that before treatment, T-bet, IFN-γ, and IL-2 were markedly down-regulated in ITP patients after treatment. Before treatment, the expressions of Foxp3, TGF-β, GATA3 and IL-4 in ITP patients did not show difference from normal controls, while after treatment Foxp3 were more up-regulated than that before treatment (p < 0.05). After treatment, TGF-β expression showed a different pattern between old and young patients. TGF-β expression was down-regulated (p < 0.05) among ITP patients younger than 60, while up-regulated in older patients. It is concluded that there is an imbalance of Th1/Th2/Treg cytokines in ITP patients, which can be reversed by glucocorticoid treatment. The conventional dose of glucocorticoid may be regarded as effective therapy for de novo ITP patients, it may correlate with improvement of imbalance between Th1/The2/Treg cytokines.
Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Forkhead Transcription Factors ; metabolism ; Gene Expression ; Gene Expression Regulation ; Glucocorticoids ; therapeutic use ; Humans ; Interferon-gamma ; metabolism ; Interleukin-2 ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; metabolism ; pathology ; T-Box Domain Proteins ; metabolism ; T-Lymphocytes, Helper-Inducer ; metabolism ; Transforming Growth Factor beta ; metabolism ; Young Adult

Objective: To observe the clinical efficacy of Zhen'ai needling method in Nei Jing (Classic of Internal Medicine) for children with allergic rhinitis (AR) accompanied by adenoid hypertrophy (AH). Methods: A total of 74 children who met the screening criteria were divided into a Zhen'ai group and a control group by the random number table method, with 37 cases in each group. The control group was treated with acupuncture at Zusanli (ST 36), Hegu (LI 4), Yingxiang (LI 20), Juliao (ST 3), Yintang (GV 29), Shangxing (GV 23) and Baihui (GV 20). The Zhen'ai group added points of Zhen'ai needling method {Shanglianquan [Extra, located at 1 cun above Lianquan (CV 23)], Tianrong (SI 17) and Lieque (LU 7)} in addition to the points in the control group. The needles were retained for 30 min. The treatment was performed twice a week. The total nasal symptom score (TNSS), sino-nasal outcome test-20 (SNOT-20) and symptom scale for AH (SSAH) were assessed before and after 10 treatments. The clinical efficacy of the two groups was compared after treatment. Results: During the treatment, 5 cases dropped out in the control group and 2 cases in the Zhen'ai group. After treatment, the total effective rate of the Zhen'ai group was 94.3％, versus 93.8％ in the control group. There was no significant difference between the two groups (P>0.05). The markedly effective rate of the Zhen'ai group was 42.9％, versus 12.5％ in the control group, and the difference was statistically significant (P<0.05). After treatment, there were significant intra-group differences in the scores of TNSS, SNOT-20 and SSAH in both groups (all P<0.05); the scores of SNOT-20 and SSAH in the Zhen'ai group were lower than those in the control group, and the differences between the groups were statistically significant (both P<0.05). Conclusion: Both conventional acupuncture and conventional acupuncture plus Zhen'ai needling method can improve clinical symptoms of children with AR accompanied by AH; and conventional acupuncture plus Zhen'ai needling method has a better effect than conventional acupuncture in improving AH symptoms.

OBJECTIVETo know the anti-viral effects of rhubarb ethanol extract (REE) on herpes simplex virus(HSV) infection in vivo.

METHODSBALB/c mice inoculated from tail vein with 0.15 ml of HSV (TCID50=10(3)) were injected hypodermically with REE next day. After divided into seven groups, three groups of mice were given different doses of REE respectively and the other groups as controls. Pathological sections from the liver, spleen, kidney were made at different times of postinfection, and their pathological changes were observed under microscope; the virus titers in viscera were assayed by using plaque formation technique and the rhubarb inhibitions to the infection of HSV in vivo?were observed.

RESULTSNo toxic response to mice were observed for REE injected hypodermically; no pathological changes were observed in different therapy groups of spleens. And those in livers and kidneys at medium- and high-dosed groups disappeared quickly. The effect of low-dosed group was equal to that of positive control group, acyclovir(ACV); the results of the titer tests showed that the virus decreased rapidly by using REE, especially in the medium- and high-dosed groups which were much more marked than the low-dosed group; Q test of the data showed that total mean value had statistical significance (F=49.1459, P<0.01); moreover there were statistical significance between therapy groups (ACV, DH1, DH2, DH3) and non-therapy groups (VC) (P<0.01 ) and between DH2, DH3 and DH1 (P<0.01); no statistical significance were found between DH1, DH2 or DH3 and ACV (P>0.05). Results show that as to the effect of decreasing the average of the total titer, rhubarb is as effective as ACV; furthermore, the medium- and high-dosed groups are superior to the low-dosed group.

CONCLUSIONSREE has significant anti-viral effect on HSV in vivo; there will be a wide application foreground of it in clinical usage.

Animals ; Antiviral Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Herpes Simplex ; pathology ; virology ; Herpesvirus 1, Human ; drug effects ; Male ; Mice ; Random Allocation ; Rheum

Objective To investigate the distribution of ghitathione-S-transferase M1 (GSTM1) and T1 (GSTT1) genes polymorphisms in Chinese population and smear-positive pulmonary tuberculosis cases of Jilin province. Methods Articles about GSTM1 and GSTT1 genes polymorphisms published before 2009 in China were searched. The study population was obtained from fourteen counties (or districts) of Jilin province, which included all cases from November, 2007 to May, 2008, totally 1120. The genotypes of GSTM1 and GSTT1 were detected by multiplex PCR technique. Results The frequencies of GSTM1 and GSTT1 'null' genotypes and combination M1-T1 'null' genotype acquired from systematic review were 54.2%, 46.8% and 26.2%, respectively, in Chinese Hans they were 53.4%, 44.9% and 25.5%, and in our research they are 57.2%, 20.4% and 13.7%, respectively. No significant differences between the frequencies of males and females as well as among that of different age groups were observed(P>0.05). The frequency of GSTM1 'null' genotype in our research is slightly higher than that in systematic review (P=0.016) , and the frequencies of GSTT1 'null' genotype and combination M1-T1 'null' genotype and are significantly lower than those in systematic review (both P<0.001). Conclusion The frequencies of GSTM1 and GSTTI 'null' genotypes were different among ethnics. The statistical difference between systematic review and our research may due to our large sample size and mostly Soutbern people in previous studies.

OBJECTIVETo investigate the expression differences of apoptosis-inducing factor (AIF) and caspase-3 among colorectal carcinoma, adenoma and normal mucosa, and to identify the relationship between AIF and caspase-3 expression in colorectal adenoma-carcinoma sequence.

METHODSFormalin-fixed paraffin embedded colorectal tissues from 174 cases, including 84 adenomas, 72 carcinomas, and 18 normal mucosa, were examined for expression of AIF and caspase-3 by streptavidin-peroxidase (SP) immunohistochemistry.

RESULTSThe positive rates of AIF and caspase-3 in colorectal adenoma were higher than those in normal mucosa (P <0.05). The positive rate of AIF in adenoma showed no significant difference compared to colorectal carcinoma (P >0.05). However, caspase-3 expression in adenomas was significantly higher than that in carcinoma (P <0.05). The positive rate of AIF in tubular adenoma was significantly higher than that in villous adenoma (P <0.05), while the positive expression rate of caspase-3 in the two types of adenoma showed no significant difference (P >0.05). AIF expression had no prominent correlation with the caspase-3 expression (P >0.05).

CONCLUSIONSThe dysregulation of caspase-independent pathway of apoptosis may be an early event in the development of colorectal carcinogenesis, while the dysregulation of the caspase-dependent pathway of apoptosis may be one of contributing factors of colorectal carcinogenesis. The caspase-independent pathway of apoptosis and the caspase-dependent pathway of apoptosis are two relatively independent pathways in colorectal carcinogenesis.

Adenoma ; metabolism ; pathology ; Adult ; Aged ; Apoptosis Inducing Factor ; metabolism ; Caspase 3 ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Intestinal Mucosa ; metabolism ; pathology ; Middle Aged ; Young Adult

BACKGROUNDRelapse happens frequently after allogeneic hematopoietic cell transplantation (allo-HCT) in the patients with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL). Detection of the minimal residual disease (MRD) before and after allo-HCT is associated with higher relapse rate. Early administration of imatinib after allo-HCT may prevent recurrent Ph(+) ALL. The aim of this study was to evaluate the safety and efficacy of imatinib in preventing hematological relapse when imatinib was administrated in the first 90 days after allo-HCT.

METHODSPatients with Ph(+) ALL that underwent allo-HCT were enrolled in a prospective study. A TaqMan-based real-time quantitative polymerase chain reaction (RQ-PCR) technique was used to detect the MRD (bcr-abl transcript levels). Imatinib therapy was initiated prior to 90 days after allo-HCT if the patient's absolute neutrophil count (ANC) was above 1.0 × 10(9)/L (without granulocyte colony-stimulating factor (G-CSF) administration) and the platelet count was greater than 50.0 × 10(9)/L, or if the bcr-abl transcript levels were elevated in two consecutive tests, or if the bcr-abl transcript levels were ≥ 10(-2) after the initial engraftment. The initial daily dose of imatinib was 400 mg/d for adults and 260 mg/m(2) for children (younger than 17 years). Imatinib was administered for at least 1 month and the bcr-abl TaqMan results were negative for 3 consecutive tests, or complete molecular remission (CR(mol)) was sustained for at least 3 months.

RESULTSFrom May 2005 to October 2008, 29 patients were enrolled in this study, of whom, 19 patients were male and 10 were female. The median age of the enrolled patients was 33 years (range 6 - 50 years). Imatinib therapy was started at a median time of 60 days (range 20 - 122 days) post HCT (only one patient started Imatinib therapy at 122nd day after HCT). Twenty-five adult patients could tolerate a dose of 300 - 400 mg/d of imatinib, and three children tolerated a dose of 260 mg×m(-2)×d(-1). Sixty-eight percent of the patients experienced various adverse events during imatinib therapy, hematological toxicity being the most common adverse event. The median duration of imatinib treatment was 3 months (range 7 days-18 months). During the median follow-up of 24 months (range 16.0 - 54.5 months), 3 out of 27 patients that could be evaluated for efficacy died from relapse. The 3-year probability of relapse for the evaluated patients was (11.3 ± 0.61)%. The relapse rates among the subgroup of positive and negative bcr-abl patients before allo-HCT were 13.6% and 0, respectively (P > 0.05). The relapse rates among the subgroups of bcr-abl positive and negative patients after allo-HCT were 20.0% and 5.9%, respectively (P > 0.05). The relapse rates among the patients in first complete remission (CR(1)) and second complete remission/non-remission (CR(2)/NR) before transplantation were 0 and 31.4%, respectively (P < 0.05). The 3-year probability of overall survival (OS) and disease-free survival (DFS) for the all enrolled patients were (75.3 ± 8.1)%. The 3-year probabilities for OS and DFS among the subgroup of patients in CR(1) and CR(2)/NR before transplantation were (87.7 ± 8.2)% and (54.6 ± 15.0)%, respectively (P < 0.05).

CONCLUSIONSAdministration of imatinib at a dose of 300 - 400 mg/d in the first 90 days after allo-HCT is feasible in Ph(+) ALL patients. With this treatment, bcr-abl positive patients before or after transplantation do not have a higher relapse rate after allo-HCT compared with the bcr-abl negative patients. Because of lower relapse rate and better OS and DFS, we recommend that Ph(+) ALL patients receive allo-HCT in CR₁.

Adolescent ; Adult ; Antineoplastic Agents ; administration & dosage ; therapeutic use ; Benzamides ; Child ; Drug Administration Schedule ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Piperazines ; administration & dosage ; therapeutic use ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; therapy ; Prospective Studies ; Pyrimidines ; administration & dosage ; therapeutic use ; Transplantation, Homologous ; methods ; Young Adult

OBJECTIVETo study the therapeutic efficacy of siRNA fragments silencing p75 neurotrophin receptor (p75(NTR)), which may be a key regulator of glioma cell apoptosis and invasion.

METHODSThe siRNA sequence fragments targeting p75(NTR) were designed and transferred into human glioma cell line U251. RT-PCR and immunocytochemistry method were used to explore the expression of p75(NTR) mRNA and protein. Cell adhesion assay was employed to detect cellular adhesion ability, and soft agar clone formation assay was adopted to identify oncogenicity, and a U251 glioma model was established in nude mice. The intracranial tumor volume was detected by MRI. The expression of p75(NTR), NGF and cyclin D2 were identified using immunohistochemistry. Cell apoptosis was detected by apoptosis kit in situ.

RESULTSThe siRNA fragments targeting p75(NTR) were capable of decreasing mRNA and protein expression of p75(NTR) in U251 glioma cell line. Both the cellular adhesion ability and oncogenicity were weakly relevant. The p75(NTR) expression level was negatively correlated with cyclin D2 and apoptosis, and positively correlated with NGF expression. The siRNA sequence fragments targeting p75(NTR) were effective in decreasing the gross volume of tumor; prolonged the survival time of mice, and the edge of tumor was much sharper than that of the control group.

CONCLUSIONSThe gene silencing technique by siRNA targeting p75(NTR) is capable of decreasing tumor invasion and cell proliferation as well as inducing cell apoptosis. It is expected to be a new choice for glioma gene therapy.

Animals ; Apoptosis ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclin D2 ; metabolism ; Gene Silencing ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Male ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Nerve Growth Factor ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Random Allocation ; Receptor, Nerve Growth Factor ; genetics ; metabolism