Mycobacterium tuberculosis L-forms(MTB-L), which has a deficient cell wall, shows various forms, various colors by acid-fast stain, osmosis pressure sensitivity and can not grow in normal isotonic culture medium, for all of the characters, misdiagnosis, underdiagnosis, difficulties of cure and recurrence became the crucial problems in the prevention and control of pulmonary tuberculosis. The recent research works on structure, biology, clinic, epidemiology and determination of MTB-L were reviewed in the present paper.

Objectives To explore the associations between drug efflux pump gene expression and phenotypic drug resistance as well as gene mutation patterns related to drug resistance of Mycobacterium tuberculosis.Methods Forty-five Mycobacterium tuberculosis isolates resistant to one or more of drugs including isoniazid, rifampicin, streptomycin and ethambutol, and 26 isolates all sensitive to the above four drugs from Tianjin Tuberculosis Control Institute in 2007 were involved in this study. Direct sequencing was applied to detect the mutations in the corresponding resistance genes(isoniazid:katG, inhA, oxyR-ahpC, ndh, rifampicin:rpoB, streptomycin:rpsL, rrs, and ethambutol:embB, embC and embA). After RNA extration and reverse transcription, real-time PCR was conducted to assess the expressions of putative drug efflux pump genes Rv1410c, Rv2136c, Rv0783c and Rv2136c, and Students' t test and ANOVA analysis were used to analyze the expression differences in Mycobacterium tuberculosis with different phenotypic drug resistance and drug resistance related gene mutation patterns.Results Compared to pan-sensitive isolates[(5.67±3.29)×10-5], Rv1410c showed higher expression in streptomycin[(8.48±6.33)×10-5, t'=2.18, P<0.05], isoniazid[(8.43±6.38)×10-5, t'=2.20, P<0.05], rifampicin[(9.59±7.27)×10-5, t'=2.29, P<0.05], multi-drug[(10.37±7.86)×10-5, t'=2.34, P<0.05] resistant isolates, and in isoniazid + streptomycin resistant isolates[(9.39±6.81)×10-5, t'=2.43, P<0.05];Rv2136c showed higher expression in isoniazid resistant[(3.51±2.43)×10-5, t'=2.03, P<0.05], multidrug-resistant isolates[(4.21±2.94)×10-5, t'=2.22, P<0.05] and resistant to isoniazid+streptomycin[(3.81±2.46)×10-5, t'=2.28, P<0.05] isolates . The expression of Rv0783c in rifampicin resistant isolates with rpoB 531 mutations [(5.41±3.03)×10-6] was higher than those with wild type of rpoB 531[(2.29±1.62)×10-6, t=2.81, P<0.05].Conclusions The expression of Rv1410c and Rv2136c are associated with mutiple-drug resistance of Mycobacterium tuberculosis.The expression of Rv0783c in rifampicin resistant isolates is associated with mutation in rpoB 531.

Objective To study the prevalence and genotype of human parvovirus B19 virus among blood products and plasma pools in China. Methods B19 DNA derived from 16 lots of factor Ⅷ concentrate produced by 4 manufactures and 10 lots of plasma pools were detected by nested PCR. Phylogenetic comparison of the partial B19 sequences obtained from positive samples were performed by direct sequencing. Results Twelve of sixteen lots of factor Ⅷ concentrate and all of ten lots of plasma pools were contaminated by B19 DNA. By comparing the partial B19 sequences,all the isolated viruses were genotype Ⅰ and their nucleotides were high conserved with homology of 98. 3%-100%. Conclusion B19 genotype Ⅰ DNA has been detected in high prevalence in factor Ⅷ concentrate and plasma pools. The genetic diversities were shown to be very low.

To obtain the functional fusion protein of rhoptry protein 2, compound rhoptry protein2 and surface antigen 1 of Toxoplasma gondii. the ROP2 and P30 genes from genomic DNA of T.gondii RH strain were amplified by PCR, and were inserted into pMD18-T cloning vector. Then the ROP2 fragment was subcloned to pET-30a(+) plasmid digested by EcoRⅠand Hind Ⅲ to construct plasmid pET-ROP2. Furthermore,the P30 fragment was subcloned into pET-ROP2 digested by BglⅡand EcoRⅠto create plasmid pET-ROP2-P30, the resulting recombinant plasmids , transformed into E.coli BL21 (DE3), were induced with IPTG. and the proteins identified by SDS-PAGE were further purified and refolded. The biological activity was analyzed by Western blot with specific antibody. It was found that the sizes of ROP2 and ROP2-P30 were 1212 and 1896bp with corresponding molecular weight 50- kDa and 75-kDa, respectively. The recombinant protein ROP2 (50-kDa) could specifically react with rabbit-polyclonal antiserum, and complex fusion protein ROP2-P30 (75- kDa) could react with P30 monoclonal antibody.

Objective To investigate the serum Ectodysplasin A(EDA)levels in women with polycystic ovary syndrome(PCOS)and explore the correlation of EDA and insulin resistance(IR).Methods A total of 256 newly diagnosed PCOS patients in Guangzhou First People's Hospital from September 2020 to September 2022 were selected.Taking BMI=25 kg/m2 as the cut-off point,the patients with PCOS were divided into obese subgroup(n=143)and non-obese subgroup(n=113).Among them,43 PCOS received six months of Liraglutide intervention.123 healthy women were normal control group(NC),which were divided into healthy obese subgroup(n=10)and healthy non-obese subgroup(n=113).Serum EDA and related biochemical indexes were detected in both groups.The correlation between serum EDA and metabolic index were analyzed.Results The serum EDA level in PCOS group was higher than that in NC group[(272.59±68.52)vs(214.51±61.36)pg/ml,P<0.05].Serum EDA in obese subgroup was higher than that in no-obese subgroup in PCOS patients[(301.11±55.81)vs(236.84±66.25)pg/ml,P<0.01].After the intervention of Liraglutide,serum EDA in PCOS women markedly decreased[(294.71±57.81)vs(336.0±61.23)pg/ml,P<0.05].EDA levels were correlated positively with body mass index(BMI),waist hip ratio(WHR),fasting blood glucose(FPG),2 hPG,fasting insulin,2 hIns,homeostasis model assessment for insulin resistance(HOMA-IR),TC,LDL-C and testosterone.Multiple stepwise regression analysis showed that BMI and HOMA-IR were independent risk factors of serum EDA.Conclusion Serum EDA is elevated in PCOS patients,especially when combined with obesity,which may be related to IR.Liraglutide intervention can significantly reduce EDA levels in PCOS patients.

OBJECTIVETo explore the role of CD4⁺ CD25⁺ Foxp3⁺ Treg/CD4⁺ IL-17A(+)Th17 cells and the related cytokines in Graves' ophthalmopathy.

METHODSBased on clinical activity scores (CAS), we divided patients with untreated Graves' ophthal- mopathy into active group (AGO group with CAS ≥ 3 (15 cases) and non-active group (NGO group) with CAS<3 (15 cases), with another 15 patients with untreated Graves' disease free of eye symptoms (GD group) and 15 normal subjects as controls. Peripheral venous blood Treg/Th17 cell ratio was determined using flow cytometry. RT-PCR was used to detect the mRNA expression levels of Treg-specific transcription factor Foxp3 and Th17-specific transcription factor RORγt. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum levels of Th17 cell-related cytokines (IL-17A, IL-23, and IL-6) and Treg-related cytokines (TGF-β, IL-10, and IL-35).

RESULTSCompared with the normal subjects, the patients in GD, NGO, AGO groups all showed significantly increased Th17 cell count (P<0.05), which was the highest in AGO group. RT-PCR results revealed significantly increased RORγt in GD, NGO, and AGO groups, also the highest in AGO group. Serum IL-17A, IL-23, and IL-6 levels all showed significant increments in GD, NGO, and AGO groups (P<0.05), especially in AGO group. Among the Treg-related cytokines, TGF-β and IL-35 levels decreased (P<0.05) but IL-10 increased significantly (P<0.05) in GD, NGO, AGO groups.

CONCLUSIONDecreased immunosuppressive capacity of Treg cells can be an important factor in the pathogenesis of Graves' ophthalmopathy. Th17 cells may also participate in the occurrence and progression of Graves' ophthalmopathy and can serve along with related cytokines as novel indicators of the disease activity. Impaired Treg/Th17 balance may importantly contribute to the occurrence of Graves' ophthalmopathy.

Cytokines ; immunology ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Graves Ophthalmopathy ; immunology ; Humans ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

Objective:To explore any effect of working memory training on executive functioning and ability in the activities of daily living after a stroke.Methods:Forty-six stroke survivors were randomly divided into an experimental group and a control group, each of 23. Both groups received conventional cognitive rehabilitation (including computer-assisted cognition training and virtual reality training using upper limb rehabilitation robots), while the experimental group also received working memory training. Before, as well as after 4 and 8 weeks of the treatment, both groups′ executive functioning was evaluated using the Trail Making test, the Frontal Assessment Battery and the Wisconsin Card Sorting Test-64. Ability in the activities of daily living was quantified using the functional independence measures (FIMs).Results:There were no significant differences between the two groups in any of the measurements before the training. After 4 weeks the average scores of the experimental group on all four instruments were significantly higher than the control group′s averages. After 8 weeks the average scores had improved significantly more. The treatment group performed significantly better than the control group throughout.Conclusions:Working memory training can improve executive functioning and the daily life of stroke survivors.