Objective To explore the association of ATP-binding cassette (ABC) efflux pump gene Rv1217c-Rv1218c and the drug resistance of Mycobacterium tuberculosis.Methods A total of 34 Mycobacterium tuberculosis clinical isolates including 24 drug-resistance isolates which were resistant to riffampicin,isoniazid,streptomycin or ethambutol and resistant to at least one second-line antituberculosis drug,and 10 drug-sensitive isolates were involved in this study.The RNA of isolated strains was extracted and then reverse transcribed. Gene expression was performed by real-time polymerase chain reaction (PCR) and data was analyzed by t test and Logistic regression analysis.Results The expressions of Rv1217c in rifampicin-resistant group (2.13 ± 1.89,t =3.44,P＜0.01),isoniazid-resistant group ( 1.84 ± 1.86,t =3.16,P＜ 0.05),streptomycin-resistant group ( 1.86 ±1.96,t=2.78,P＜0.05) and ethambutol-resistant groups (3.36±2.35,t=3.04,P＜0.05) were all higher than sensitive isolates (0.42 ± 0.31).The expressions of Rv1218c in rifampicin-resistant group (2.54±1.84,t=3.82,P＜0.01),isoniazid-resistant group (2.34± 1.84,t=3.72,P＜0.01),streptomycin-resistant group (2.15±1.86,t=3.01,P＜0.01) and ethambutol-resistant groups (3.78± 1.78,t=4.22,P＜0.01 ) were all higher than sensitive isolates (0.65 ± 0.42).The expressions of Rv1217c and Rv1218c in multidrug resistant group were 2.74±2.07 and 3.33± 1.77,respectively,which were both higher than polydrug resistant group (0.79 ± 0.47 and 1.03 ± 0.79,respectively; t =2.91,P＜0.05 ; t =3.84,P＜0.01,respectively).Logistic regression analysis found that high Rv1217c expression was positively correlated with rifampicin resistance and negatively correlated with isoniazid resistance (both P＜ 0.01 ),while high Rv1218c expression was negatively correlated with rifampicin resistance and positively correlated with isoniazid resistance (both P＜0.01 ).High expressions of two genes were both positively correlated with ethambutol resistance and multidrug resistance (both P＜0.01 ) and not statistically correlated with streptomycin resistance (P＞0.05).Conclusions The expressions of ABC efflux pump gene,Rv1217c-Rv1218c,are correlated with multiple drug resistance.The overexpression may contribute to the multidrug resistance of Mycobacterium tuberculosis.

Background and purpose:Coventional radiation has been used for decades,but the 3D dose distribution has not been studied as yet.In this study,different treatment plans of conventional irradiation technique for esophageal carcinoma have been evaluated by 3-D TPS.Methods:Five patients with esophageal carcinoma at upper-thoracic and five patients at middle-thoracic were enrolled in this study.Three conventional treatment plans were created for each patient,with a prescribed dose of 70 Gy.For tumor at upper-thoracic,plan 1 consisted of one anterior field and bilateral posterior oblique fields with wedges,plan 2 consisted of anterior bilateral oblique fields with wedges ,plan 3 consisted of a pair of AP-PA portals,then followed by right anterior oblique portals and left posterior oblique portals to spare spine cord.For tumor at middle thoracic,plan 1 consisted of one anterior field and bilateral posterior oblique fields.Plan 2 consisted of a pair of AP-PA portals and followed by another pair of parallel-opposed lateral off- cord fields as boost.Plan 3 used the same plan as for the lesion at upper-thoracic.The evaluation of each treatment plan was carried out by dose-volume histogram(DVH).Results:For tumor at upper-thoracic,on average,the maximum dose to spinal-cord in plan 3 was (57.1?4.9)Gy,and was higher than that in plan 1 and plan 2.Plan 1 increased mean lung dose from (12.8?2.1)Gy to (18.2?4.1)Gy(P=0.045)compared with plan 2,but it improved the homogenous dose of PTV2,especially in the patient with long tumor.For tumor at middle-thoracic,on average,plan 2 increased mean lung dose from (11.9?1.1)Gy to (13.0?0.6)Gy(P=0.045) compared with plan 3.Plan 2 increased V_(20) and V_(30) from (23.6?2.3)% to (29.2?1.9)%(P=0.004) and (13.9?2.3)% to (20.9?1.3)%(P=0.006) compared with plan 1.The trachea volume of(?)70 Gy(V_(70))in plan 3 was larger than that in plan 1[(20.3?15.9)% VS (10.5?9.8)%,P=0.058].Conclusion:For tumor at upper-thoracic,plan 1 and 2 were superior to plan 3.Irradiation to lung in plan 2 was lower than that in plan 1,but plan 1 improved the homogenous dose of PTV2 compared with plan 2,especially for patient with longer lesion.For tumor at upper-thoracic,plan 2 increased irradiation dose to lung compared with plan 1 and plan 3.Plan 1 was comparable with plan 3,but the dose of trachea in plan 1 may lower than that in plan 3.

OBJECTIVETo assess the response in THP-1 treated with Rv3671c protein in Mycobacterium tuberculosis (M.tuberculosis).

METHODSThe gene encoding Rv3671c protein of M.tuberculosis was cloned into pET-28a vector and then expressed in Escherichia coli. The Rv3671c was purified with Ni-NTA affinity and ion exchange chromatography. The detection of protein concentration was by Lowry method.THP-1 cell was stimulated with Rv3671c protein and cells were analyzed by Hochest staining under fluorescence microscopy to assay cell death (apoptosis and necrosis). TNF-α and IL-1β were detected by ELISA at each stimulating time.

RESULTSThe Rv3671c protein of M.tuberculosis was successfully expressed in Escherichia coli. The purity of recombinant Rv3671c protein was 95%, and the protein concentration was up to 0.4 mg/ml. The nucleus of THP-1 was isolated and necrosis-like under fluorescence when cells were stimulated by Rv3671c protein. The levels of TNF-α and IL-1β in supernatant were 19 000 and 16 500 pg/ml respectively, and were significantly higher than control cells with the levels of 2100 and 3800 pg/ml separately.

CONCLUSIONThe necrosis of THP-1 cells could be stimulated by Rv3671c protein of M.tuberculosis and it was probably associated with high cytokines TNF-α and IL-1β levels.

Bacterial Proteins ; pharmacology ; Cell Death ; Cell Line ; Humans ; Interleukin-1beta ; metabolism ; Macrophages ; cytology ; metabolism ; Mycobacterium tuberculosis ; genetics ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo evaluate the effect of portal hypertension on prognosis in patients with decompensated liver cirrhosis.

METHODSThe clinical data of decompensated cirrhosis patients in our hospital, between 2003 and 2006, were retrospected and followed up. Model for end-stage liver disease (MELD) score and Child-Turcotte-Pugh (CTP) classification was calculated using the standard formula. Kaplan-Meier survival analysis was used to compare the mortality in subgroups ranked by the syndromes. Cox proportional hazards regression was used to evaluate the effect of the syndromes on prognosis.

RESULTSA cohort of 322 patients was admitted in this study at the end of the follow-up. The mortality of variceal bleeding, hepatic encephalopathy, a large volume ascites, spontaneous bacterial peritonitis, the type I and type II hepatorenal syndrome was 45.9%, 79.4%, 66.7%, 100%, 100% and 84.6% respectively. On the whole, the occurrence of all the syndromes was correlated with CTP classification and MELD score. Kaplan-Meier survival analysis showed that all of these syndromes, except for low to medium volume of ascites, significantly affected the survival rate (P<0.01). In Cox regression analysis, all the syndromes were the independent predictors of prognosis, the regression coefficient values of hepatic encephalopathy, spontaneous bacterial peritonitis, type I and type II hepatorenal syndrome, variceal bleeding and ascites were 0.973, 0.928, 0.935, 0.866, 0.464 and 0.369 respectively.

CONCLUSIONSThe portal hypertensive syndromes have significant effect on the prognosis of the patients with decompensated cirrhosis, hepatic encephalopathy is the worst one.

Adult ; Aged ; Esophageal and Gastric Varices ; epidemiology ; etiology ; Female ; Follow-Up Studies ; Gastrointestinal Hemorrhage ; epidemiology ; etiology ; Hepatic Encephalopathy ; epidemiology ; etiology ; Hepatorenal Syndrome ; epidemiology ; etiology ; Humans ; Hypertension, Portal ; complications ; epidemiology ; Liver Cirrhosis ; complications ; mortality ; pathology ; Male ; Middle Aged ; Models, Statistical ; Predictive Value of Tests ; Prognosis ; Severity of Illness Index ; Survival Analysis

Objective:To investigate the changes in peripheral blood and liver-infiltrating natural killer-like B (NKB) cells in patients with primary hepatocellular carcinoma (HCC), and to assess the influence of IL-18 on NKB cells in vitro and the underlying mechanism. Methods:Forty-three HCC patients and 21 normal controls (NC) were enrolled in the study. Peripheral blood samples were collected to isolate plasma and peripheral blood mononuclear cells (PBMC). Intrahepatic lymphocytes (IHL) were isolated from tumor tissues and para-tumor tissues obtained from 16 HCC patients who received surgery. IL-12, IL-18 and IL-18 binding protein (IL-18BP) levels in plasma were measured by enzyme linked immunosorbent assay. The percentages of CD3 -NKp46 + CD19 + NKB cells and IL-18 + NKB cells in PBMC and IHL were analyzed by flow cytometry. Changes in the percentages of NKB cells and IL-18 + NKB cells were measured after stimulating PBMC and IHL with recombinant human IL-18 (1 ng/ml and 10 ng/ml). Changes in IL-18BP levels in the culture supernatants and phosphorylated nuclear factor-κB (NF-κB) in NKB cells were also assessed. Student′s t test, one-way analysis of variance or LSD-t test was used for statistical analysis. Results:There was no significant difference in plasma IL-12 level between HCC patients and NC ( P=0.245). Compared with NC, HCC patients had decreased IL-18 level in plasma [(224.3±58.89) pg/ml vs (327.0±52.27) pg/ml, P<0.000 1], but increased IL-18BP level [(4.421±0.97) ng/ml vs (0.92±0.18) ng/ml, P<0.000 1]. The percentages of peripheral blood NKB cells and IL-18 + NKB cells were lower in HCC patients than in NC [(2.68±1.23)% vs (8.88±2.95)% and (54.42±12.60)% vs (69.74±12.65)%, both P<0.000 1]. The percentage of NKB cells in IHL was reduced in tumor tissues as compared with that in para-tumor tissues [(2.89±0.86)% vs (4.66±1.17)%, P<0.000 1]. Moreover, the percentage of IL-18 + NKB cell was also down-regulated in tumor tissues as compared with that in para-tumor tissues [(51.50±13.18)% vs (62.13±9.24)%, P=0.013]. Recombinant human IL-18 stimulation reduced the IL-18BP level in the culture supernatants ( P<0.05). IL-18 stimulation at 1 ng/ml did not affect NKB cell percentage, IL-18 + NKB cell percentage or NF-κB phosphorylation in NKB cells from PBMC or IHL ( P>0.05), while 10 ng/ml of IL-18 not only elevated NKB cell percentage and IL-18+ NKB cell percentage, but also promoted NF-κB phosphorylation in NKB cells ( P<0.01). Conclusions:In vitro stimulation with high concentration of IL-18 might promote NF-κB phosphorylation by inhibition of IL-18BP expression. This process might play a positive feedback role to induce the activation of NKB cells and IL-18 secretion.

Objective To explore the effects of repressor element 1-silencing transcription factor (REST)/REST coinhibitory factor (CoREST) on axonal regeneration and repairmen of mouse cortical neurons after traumatic brain injury (TBI).Methods (1) The primary cortical neurons were obtained from fetal C57BL/6 mice;one,three,5,7,9,and 11 d after cultivation,miR-124 expression was detected by quantitative-(q-) PCR.(2) Neurons cultured for 5 d were divided into miR-124 mimics group,blank control group,and miR-124 inhibitor group,and miR-124 mimics,nonsense control sequences and miR-124 inhibitor were transfected,respectively;0,6,12,24,48,and 72 h after transfection,miR-124 expression was detected by q-PCR.(3) Neurons cultured for 7 d were divided into blank control group Ⅰ,oxygen glucose deprivation (OGD) model group,up-regulated miR-124+OGD model group,and down-regulated miR-124+OGD model group,and neurons in the later two groups were transfected with miR-124 mimics and miR-124 inhibitor;48 h after transfection,OGD models in the later three groups were prepared;0,6,12,24,48,and 72 h after OGD,miR-124 expression was detected by q-PCR;GAP-43,REST and CoREST expressions were detected by Western blotting 48 h after OGD;the REST and CoREST expressions were measured by immunofluorescent staining 48 h after OGD.Results (1) One,three,5,and 7 d after cultivation,miR-124 expression gradually increased,and 7,9,and 11 d after cultivation,miR-124 expression gradually decreased,with significant differences (P＜0.05).(2) Twenty-four and 48 h after transfection,miR-124 expression in the miR-124 inhibitor group was significantly lower than that in the blank control group (P＜0.05);12,24,48 and 72 h after transfection,miR-124 expression in the miR-124 mimics group was significantly higher than that in the blank control group (P＜0.05),and peak level was noted at 48 h.(3) The miR-124 expression in the OGD model group was significantly higher than that in the blank control group Ⅰ at 12,24,48 and 72 h after OGD (P＜0.05),and peak level was noted at 48 h;0,6,12,24,48,and 72 h after OGD,the miR-124 expression in the up-regulated miR-124+OGD model group was significantly higher than that in the blank control group Ⅰ (P＜0.05),and peak level was noted at 48 h;Western blotting indicated that GAP-43 and CoREST gradually increased,and REST gradually decreased in blank control group Ⅰ,OGD model group and down-regulated miR-124+OGD model group,with significant differences (P＜0.05);neurons in the up-regulated miR-124+OGD model group had significantly lower GAP-43 and CoREST expressions,and significantly higher REST expression than those in the OGD model group (P＜0.05);the results of immunofluorescence staining were consistent with those of Western blotting.Conclusion REST/CoREST,as a pair regulator,may play a key role in the repairment and regeneration of neuron axons after TBI.

Objective: To study the mechanism and significance of pH change in the coronary artery microthrombosis of rats. Methods: After the sodium laurate-induced model of coronary artery microthrombosis of rats was constructed, the vascular endothelial cells were separated and then cultured in the mediums with different pH values for 24 h. Enzyme linked immunosorbent assay was used to detect the content of von Willebrand factor (vWF) in the medium; while the real-time PCR and western blot assay were used to detect the expression of fibrinogen-like protein 2 (FGL2) at the mRNA and protein level. The comprehensive evaluation was performed to discuss the effect of pH change on the coronary artery microthrombosis of rats. Results: The expression level of vWF detected by enzyme linked immunosorbent assay was 336.67 ± 24.95, 311.33 ± 14.98, 359.67 ± 39.63, 354.67 ± 49.01 and 332.00 ± 33.42 (pg/mL) respectively; while the expression of vWF in the model group was 570.00 ± 57.94, 524.67 ± 57.94, 437.00 ± 95.38, 415.33 ± 44.38 and 444.67 ± 74.31 respectively. Being cultured under the different pH values, the relative expression level of FGL2 mRNA in the model group was 7.93 ± 0.93, 6.70 ± 0.70, 5.03 ± 0.32, 5.13 ± 0.40 and 5.57 ± 0.83 respectively. Conclusions: The coronary artery microthrombosis of rats can cause the high expression and secretion of vWF. Meanwhile, FGL2 is also up-regulated in the thrombosis and such up-regulation is more significant in the condition with low pH, which indicates that the low pH condition may be one of factors that contribute to the cardiovascular diseases.

Objective: To analyze the difference in blood uric acid levels between patients with polycystic ovary syndrome (PCOS) and healthy women of childbearing age, and to investigate the correlation between body composition and blood uric acid levels. Methods: A total of 153 eligible childbearing age patients with PCOS treated at Tianjin Medical University General Hospital from January 2018 to March 2022 were selected, and 153 healthy women with normal menstruation were selected as the control group. Fasting blood uric acid levels were measured by venous blood test, and body composition was measured by a body composition analyzer. Group comparisons were made to analyze the correlation between body composition and blood uric acid levels. Results: The incidence of hyperuricemia was higher in patients with PCOS than that in the control group [30.1% (46/153) vs 2.0% (3/153)], with a statistically significant difference (χ²=44.429, P<0.001). Blood uric acid level was also significantly higher in patients with PCOS than that in the control group [(371±98) vs (265±67) μmol/L; t=11.170, P<0.001]. Among PCOS patients, there were statistically significant differences in weight, body mass index (BMI), body fat mass, skeletal muscle mass, percent body fat, lean body weight, fat mass/lean body weight, percent skeletal muscle, and visceral fat level between the hyperuricemia group and the normal blood uric acid group (all P<0.001), but no significant difference was observed in waist-hip ratio (P=0.348). The following body composition indicators: weight, BMI, waist-hip ratio, body fat mass, skeletal muscle mass, percent body fat, visceral fat level, lean body weight, and fat mass/lean body weight in all subjects, the PCOS patients and the control group, were positively correlated with blood uric acid levels (all P<0.01). The blood uric acid level in PCOS obese patients was higher than that in non-obese PCOS patients, and the difference was statistically significant [(425±83) vs (336±91) μmol/L; t=6.133, P<0.001]. The blood uric acid level in central obesity PCOS patients was also higher than that in non-central obesity PCOS patients [(385±95) vs (299±79) μmol/L], the difference was statistically significant (t=4.261, P<0.001). The blood uric acid level in normal-weight obese PCOS patients was higher than that in normal-weight non-obese PCOS patients [(333±73) vs (277±54) μmol/L], and the difference was statistically significant (t=2.848, P=0.006). Blood uric acid levels in normal-weight [(315±74) vs (255±67) μmol/L], overweight [(362±102) vs (276±57) μmol/L], and obese PCOS patients [(425±83) vs (303±74) μmol/L] were all higher than those in the corresponding control groups, with statistically significant differences (all P<0.001). Conclusions: PCOS patients have a higher incidence of hyperuricemia than healthy women of childbearing age. Blood uric acid levels are closely correlated with body composition indicators, such as weight, BMI, waist-hip ratio, body fat mass, skeletal muscle mass, percent body fat, and visceral fat level. Body composition analysis of women with PCOS could help identify potentially obese people more accurately and carry out individualized treatment, thereby reducing the risk of metabolic abnormalities.
Humans ; Female ; Polycystic Ovary Syndrome/complications* ; Uric Acid ; Hyperuricemia/complications* ; Insulin ; Body Composition/physiology* ; Obesity/complications* ; Body Mass Index