Objective To analyze the risk factors of pulmonary non-tuberculous mycobacterial (NTM) infection in sputum acid-fast bacilli positive and/or mycobacteria culture positive patients.Methods One hundred and sixty-three patients with pulmonary NTM infection were recruited from Jan 2006 to Jun 2011 in Shanghai Pulmonary Hospital and 326 patients with sputum positive pulmonary tuberculosis who were selected by random systemic sampling method in the same period were recruited as control.The data were retrospectively analyzed.The related factors were compared between groups by chi-square test.The risk factors of pulmonary NTM infection were analyzed by binary Logistic regression model.Results There were statistically significant differences of age,history of smoking,chronic obstructive pulmonary disease (COPD),bronchiectasis,thin walled cavities focus,purified protein derivative (PPD) test between patients with pulmonary NTM infection and patients with pulmonary tuberculosis (all P＜0.05).Univariate analysis showed that age [45-60year (OR=2.637,95％CI:1.631-4.264; P＜0.001); ＞60 year (OR=4.194,95％CI:2.581-6,813 ; P＜0.001)],history of smoking [10-20 year (OR=1.842,95％CI:1.0843.070; P=0.024),＞20 year (OR=2.040,95％CI:1.167-3.567; P=0.012)],COPD (OR=2.698,95％CI:1.588-4.583; P＜0.001),bronchiectasis (OR=3.566,95％CI:2.343-5.427;P＜0.001),thin walled cavities focus (OR=2.592,95％CI:1.581-4.250; P＜0.001) and a weak-positive reaction of PPD test (OR=2.389,95％CI:1.276-4.472; P=0.006) were all risk factors of pulmonary NTM infection.Multivariate analysis showed that age＞60 year (OR=3.961,95％CI:2.183-7.189 ; P＜0.001),bronchiectasis (OR =3.880,95 ％ CI:2.342-6.487 ; P＜0.001),thin walled cavities focus (OR=2.898,95％CI:1.567-5.360; P＜0.001),COPD (OR=2.503,95％ CI:1.289-4.857; P=0.007),age45-60 year (OR=2.452,95％CI:1.391 4.325; P=0.002)anda weak-positive reaction of PPD test (OR=2.295,95％CI..1.132-4.652; P=0.021) were independent risk factors of pulmonary NTM infection.Conclusion In sputum acid-fast bacilli positive and/or mycobacteria culture positive patients,age≥ 45 year,COPD,bronchiectasis,thin walled cavities focus and a weak-positive reaction of PPD test are risk factors of pulmonary NTM infection.The clinicians should pay close attention to the results of species identification.

Objective To evaluate cerebrospinal fluid adenosine deaminase(CSF-ADA)activity in the diagnosis of tuberculous meningitis(TMB), and to observe its dynamic changes. Methods A total of 160 patients were included and were divided into two groups: 76 cases of TBM and 84 cases of non-TBM.Among the cases of non-TBM, there were 36 cases of bacterial meningitis, 30 cases of viral meningitis and 18 cases of cryptocoocal meningitis. All the patients were measured with their CSF-ADA activity by Enzymecoupled assay(Trinder method)and 47 patients of TBM were measured again after 2 weeks' and 6 weeks'antitubercular therapy. Results were expressed as(-x)± s. Mann-Whitney U test and paired-samples t test were used. Results CSF-ADA activity in TBM group was(12.9 ±6.4)U/L, while that in the non-TBM group was(6.0 ± 4.1)U/L, the difference was of statistical significance(U = 7.860, P ＜ 0.05). With the cutoff value of 9 U/L, the sensitivity and specificity to differentiate TBM from non-TBM was 84.21% and 83. 33%, respectively. CSF-ADA activity decreased in TBM patients after antitubercular treatment.Conclusions CSF-ADA activity can be an effective laboratory marker for early differential diagnosis of TMB with the cut-off value of 9 U/L. Dynamic changes of CSF-ADA activity may be a indicator for the effect of antitubercular treatment.

Objective To explore the incidence and risk factors of anti-tuberculosis (TB) drugs induced liver injury (ATDILI) and to discuss its impact on the treatment outcome of patients treated with first line anti-TB drugs.Methods Among the patients who received anti-TB treatment with directly-observed treatment strategy (DOTS),121 patients with ATDILI and 817 patients without ATDILI were included in this retrospective cohort study.Binary Logistic regression model was used to analyze the risk factors of ATDILI in univariate and multivariate analysis.The x2 test was used to compare the treatment success rates and drug resistant rates.Kaplan-Meier analysis and Log-rank test were used to compare the sputum smear/culture conversion rates and cavity closure rates.Results The incidence of ATDILI was 12.9％ (121/938) in this cohort.Multivariate Logistic regression showed that hepatitis B virus carrier with both hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive (OR=4.29,95％CI:2.15-8.58,P＜0.01),complicated with systemic lupus erythematosus (OR=3.34,95％CI:1.46-7.63,P=0.004),serum albumin ≤25 g/L (OR=3.14,95％CI:1.50-6.58,P=0.002) and alcoholism (OR=1.79,95％CI:1.14-2.82,P=0.012) were independent risk factors of ATDILI.The treatment failure rate in patients with ATDILI was significantly higher than that in patients without ATDILI (19.1％[24/121] vs8.0％[65/817],OR=2.86,95％CI:1.71-4.78,P＜0.01).The drug resistant rates of two groups were not significant different (4.1％[5/121] vs 1.7％[14/817],P＞0.05).The sputum smear/culture conversion rate (85.4％[41/48] vs 94.0％ [298/317],x2 =38.912,P＜0.01) and cavity closure rate (84.6％[22/26] vs 93.0％[198/213],x2 =20.709,P＜0.01) in patients with ATDILI were both significantly lower than those in patients without ATDILI.Conclusions The incidence of ATDILI is relatively high in hospitalized patients treated with first line anti-TB drugs.ATDILI has negative effects on treatment outcome of TB patient.Hepatitis B carrier with positive HBsAg and HBeAg,systemic lupus erythematosus,albumin ≤25 g/L and alcoholism may increase the risk of developing ATDILI.

In this study ,the objective is to establish a nested‐PCR assay for the detection of H .bilis with high sensitivity and specificity .The nested primers were designed based on sequences of 16S rRNA gene of seventeen subtypes of H .bilis .Af‐ter optimizing reaction condition ,the sensitivity and specificity of the assay were examined via the detection of feces simulated samples ,mice infection model samples and clinic patients’ samples .The detection sensitivity of H .bilis strain for feces simu‐lated samples was 10 CFU/100 μL .H .bilis was successfully detected in the liver ,caecum and feces of experimentally infected mice .Moreover ,H .bilis was successfully detected in the bile ,cholecyst mucous membrane and feces samples from two of ten patients with cholelithiasis .Due to the PCR assay’s high sensitivity and specificity ,the method may be used to detect the infec‐tion of H .bilis .

Objective To investigate the immunomodulatory effect of mesenchymal stem cells (MSCs) and their role in prolonging allograft survival in rat heart transplantation. Methods Inbred Wistar rats were used as donors, and Fisher 344 as recipients. MSC were isolated from femur and tibia bone marrow of donors and cultured in vitro. Mixed lymphocyte reaction assays were performed to assess the immunosuppressive effects of different concentrations of MSC on allogeneic T cell proliferation. Cardiac allograft model was established and according to different intervention measures recipients were divided into two groups (MSC treatment group and control group) (n=8 in each group). In MSC treatment group, recipients were infused with 2×106 MSC via the tail vein at designated intervals (one week before operation, during operation and consecutive three days postoperation), while in control group, the recipients were treated with Ringer's solution at the same interval& At day 5 posttransplantation real-time PCR was used to detect the changes in the expression of Thl and Th2 cytokine genes in transplanted hearts. Results In vitro allogeneic T cell response was greatly suppressed by MSC in a dose-dependent manner. Real-time PCR revealed that IL-1β,IFN-γ, IL-4 and IL-10 were expressed in MSC treatment group, while IL-4 and IL-10 were not expressed in control group but with significantly higher expression of IL-1β and IFN-γ. As compared with control group, survival of MSC-treated allografts was markedly prolonged as compared with control group (mean survivaldays: 12.4±5.3 vs 6.4±2.0, P＜0.05). Conclusion Intravenous adrninistmtion of MSC can prolong the survival of transplanted heart possibly by induction of allograft tolerance through changing Th1/Th2 balance.

Objective To analyze risk factors for metabolic syndrome in Mongolia versus Han nationalities in Xilinhaote city. Methods Using the epidemiology investigation data of health examination,we calculated the prevalence of metabolic syndrome of Mongolia and Han nationalities, then used logistic regression model to explore risk factors of two nationalities. Results The crude prevalence of MS in Mongolia and Han nationality was 34.3%and 24.6% respectively. The multivariate logistic regression showed that male, the meat-rich diets and aging(OR:2.18, 1.92, 1.04 respectively)were the risk factors for Mongolia nationality, and smoking, family history of hypertension, drinking, the meat-rich diets, aging(OR:1.89, 1.84, 1.72, 1.61 and 1.04 respectively)were the risk factors for Han nationality. Conclusions Xinlinhaote population has higher MS prevalence, and different nationalities have different risk factors. We should take preventive actions to control it.

Objective To explore the correlations of adult attachment with interpersonal conflict resolu-tions styles and differences between soldiers and employees. Methods Relationship questionnaire (RQ),experi-ence in close relationships (ECR),and Rahim handling interpersonal conflict inventory (ROCI-II) were adminis-tered to 543 employees from different enterprises in Beijing and Liaoning province and 701 soldiers. Results The results suggested the distribution of adult attachment style between soldiers and employees were significantly differ-ent. Compared with employees, there were higher dismissing hat lower secure rate in soldier sample. On handling interpersonal conflict strategy indices, such as integrating, avoiding, dominating and obliging but not compromising (t_((1120))=1.97, P<0.05), there were no significant differences between soldiers and employees. Only on in-tegrating, there were significant ditterenees among four adult attachment styles (F_((3,1009))=10.18, P<0.001).Attachment avoidance had a significantly negative correlation with integrating, obliging and compromising (r=-0.31,-0.11,-0.13 respectively), while attachment anxiety had moderate but significant correlations with a-voiding, dominating and obliging and compromising (r=0. 16,0.35, 0.35, 0.39, 0.30 respectively). Conclu-sion Adult attachment is closely related to handling interpersonal conflict strategies, and there was difference on compromising between soldiers and employees.

Purpose To investigate the clinicopathologic characteristics, differential diagnosis, treatment and prognosis of differentia-ted-type vulvar intraepithelial neoplasia ( dVIN) . Methods Clinicopathologic findings and immunophenotypes of 6 cases diagnosed as“dVIN” were retrospectively analyzed, and the relevant literatures were also reviewed. Results 6 patients were all female ranged 53~80 years old with mean age of 62 years old. Clinical aspects included leukoplakia vulvar, pruitis, irritation, pain, ulcer and so on. The histopathological features were hyperplasia of basal and parabasal layer with elongation and anastomosing reteridges. Cells were marked atypia with obvious nucleoli, atypical mitosis, and dyskeratosis. In the middle and surface layer, cells were well differentiated with pronounced intercellular bridges, and eosinophilic cytoplasm, hyperkeratosis and parakeratosis. Oedema and band of infiltration of chronic inflammatory cells of subepidermal could been seen. Immuohistochemistry showed the expression rates of p53 and p16 in totally 6 cases were 83. 3% (5/6), 0 (0/6), respectively. The Ki-67 index was more than 90% in basal and parabasal cells. Four patients were followed up ( mean follow-up 17 months, range 6~36 months) , one patient died at 9 months later after surgery, another patient recurred at 6 months later after surgery, both of the 2 cases were all with invasive lesions after resection, and the rest two cases had no recur in 18 months and 36 months after surgery, respectively. Conclusion dVIN is a high grade squamous intraepithelial lesions of vulvar with low incidence rate, but had more risk of progression. p53, p16 and Ki-67 stain were useful in the diagnosis of dVIN.

Objective To clone ,construction ,express and purify Tp47 of Treponema pallidum (Tp) ,and assess the immunoreac‐tivity by Western‐Blot .Methods Tp47 was amplified by polymerase chain reaction ,and then cloned to the vector pGEX‐6P‐1 .The correct sequence of the recombinant plasmids pGEX‐6P1‐Tp47 was transformed into Escherichia coli BL21 (DE3) and induced .The expression product was analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western‐Blot .The expression protein was purified .Serum of different clinical stages of syphilis was used as the antibody to detect the immunoreactivity of the protein by Western‐Blot .Results A fusion protein with molecular weight about 71 × 103 was attained .Western‐Blot proved that the recombinant protein can react with Tp IgG positive sera .And the specificities and sensitivities of the diagnostic reagent detected by sera were 100% .Conclusion The recombinant protein Tp47 was expressed and purified with good antigen activity ,which could provide the basis of theory and practice for the development of early diagnostic kit applying to detect Tp infection .

OBJECTIVETo investigate the difference of the transcripts between reticulocyte and non-reticulocyte cells in human blood.

METHODSGenomic DNA, reticulocyte RNA and total RNA of K-, K+ and Kell-null(K0) were extracted, then PCR, reverse transcription-PCR(RT-PCR) and nested PCR followed by sequencing or cloning-sequencing were used to analyze the KEL gene mRNA exons 1-19 and exons 2-8. Four kinds of monoclonal antibodies were labeled to detect the expression of Kell glycoprotein on red cells or leukocytes with flow cytometry.

RESULTSIn reticulocyte, only one normal KEL transcript faithful to the genomic structure was found in all tested samples except K0 which had 4 different transcripts. Sequence analysis of exons 2-8 of total RNA confirmed the alternative KEL transcripts existed in different samples, mostly caused by abnormal splicing, among them, skipping of exon 3 and a 16 bp insertion of intron 6 at the beginning of exon 7 were the most frequent. Although only one band was observed after amplifying the exons 1-19 from total RNA, the sequencing result showed it was a mixture of different sequences. There was strong expression of Kell glycoprotein on red cells except K0, but no or low expression on leucocytes by flow cytometry.

CONCLUSIONAlternative transcripts of KEL gene exist in different cells, which would be responsible for different Kell glycoprotein expression patterns on different cells. This study suggested that reticulocyte RNA was more suitable than total RNA for molecular study of KEL gene transcription.

Base Sequence ; Cloning, Molecular ; DNA ; genetics ; Exons ; genetics ; Genome, Human ; Genomics ; Humans ; Introns ; genetics ; Kell Blood-Group System ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; blood ; genetics ; Reticulocytes ; cytology ; metabolism ; Sequence Analysis, DNA