Objective To investigate the change of 25-hydroxyl vitamin D level and its impact on glucose metabolism and bone mineral density in type 2 diabetic patients.MethodsTwo hundred and fifty-eight cases of type 2 diabetic patients in our hospital were collected.In accordance with 25 (OH) D =50 nmol/L for the critical values,they were grouped into vitamin D deficiency group ( n =192) and the relative lack of vitamin D group ( n =66).Dual-energy X-ray absorptiometry (DXA) measurement of the lumbar vertebrae L2 - L4 and femoral neck bone mineral density were undertaking.Taking the total value of the lumbar spine,femoral neck and total value as judgment indicators,homeostasis model assessment of insulin resistance (HOMA-IR),we collected and statistically analyzed glucose metabolism and bone metabolism indicators in patients with diabetes.Results Among 258 cases of type 2 diabetes,there were 57 cases of osteoporosis accounting for 22.1％.There was significant difference on the duration of diabetes [ (7.98 ± 1.09 ) years old vs (3.77 ± 1.21 ) years old,t =4.849,P ＜0.05],FINS [ (6.42 ± 1.30) mU/L vs (5.79 ± 1.08) mU/L,t =3.871,P ＜0.05] and HOMA-IR [ (2.35 ±0.54) vs ( 1.85 ±0.41 ),t =2.705,P ＜0.05],but no significant difference on the FPG and HbA1c ( P ＞ 0.05 ) between two groups.PTH [ ( 36.51 ± 7.59) ng/L vs ( 32.02 ± 6.89 ) ng/L,t =2.008,P ＜ 0.05 ],bone mineral density in the lumbar total value [ (0.87 ±0.14) g/cm2 vs (0.99 ±0.12) g/cm2,t =2.799,P ＜0.05 ],femoral neck Neck [ ( 0.70 ± 0.10 ) g/cm2 vs ( 0.79 ± 0.11 ) g/cm2,t =2.564,P ＜ 0.05 ] and Total [ (0.84 ± 0.14) g/cm2 vs (0.97 ± 0.15 ) g/cm2,t =3.340 P ＜ 0.05 ] were statistically different,but no significant difference on the rest of the indicators.Conclusion The prevalence of vitamin D deficiency was found in patients with diabetes.Vitamin D level could affect insulin resistance and glycemic control and bone mineral density level.Therefore,routine vitamin D testing to the diabetic patients and giving vitamin D replacement therapy to vitamin D deficiency patients in a timely manner were necessary.

To observe BAG-1, EGFR, and PARP-1 expressions in invasive breast cancer and its correlation with clini-cal pathological indicators, as well as to evaluate their clinical significance. Methods:The BAG-l, EGFR, and PARP-1 expressions in a tissue microarray of invasive breast cancer and peritumoral tissues were detected through immunohistochemical staining. The clinical and pathological significance of BAG-1, EGFR, and PARP-1 were evaluated. Results:The BAG-1, EGFR, and PARP-1 expression lev-els are higher in invasive breast cancer tissues than in peritumoral tissues (P<0.05). BAG-1 expression in invasive cancer tissues is not related to age, tumor site, lymph node metastases, and clinical TNM staging of patients, but is related to size, grade, ER, PR, and HER-2 expressions and molecular subtype (P<0.05). EGFR expression is related to size, clinical TNM staging, and molecular subtype (P<0.05). PARP-1 expression is related to grade, lymph node metastases, ER, and molecular subtype (P<0.05). BAG-1 expression is not significantly correlated with EGFR and PARP-1 in all cases, but BAG-1 and PARP-1 expressions are positively correlated in tri-ple-negative breast cancer tissues (P<0.05). Results of the univariate analysis revealed that the BAG-1 and PARP-1 expressions and the molecular subtypes are associated with the prognoses of breast cancer patients. Multivariate analysis revealed that BAG-1 and PARP-1 expressions are factors that are independent of the prognosis. Conclusion: BAG-1, EGFR, and PARP-1 overexpressions in human breast tissues suggest that BAG-1, EGFR, and PARP-1 are related to breast cancer development. BAG-1, EGFR, and PARP-1 are poten-tial biomarkers of breast cancer diagnosis and prognosis.

Objective To observe the changes of endothelin-1 (ET-1) and prostaglandin E2 (PGE2) in exhaled breath condensate (EBC) and serum of patients with acute respiratory distress syndrome (ARDS) after treated by Qingfeitang and investigate its clinical value. Methods 52 ARDS patients receiving mechanical ventilation at intensive care unit (ICU) were divided into the Qingfeitang treatment group and the control group, with 26 cases in each group. The EBC were collected by Ecoscreen condenser within 24 h after diagnosis of ARDS and on the 5th day of medication, and the venous blood were collected at the same time. The levels of ET-1 and PGE2 in the EBC and serum of different period were measured by EIA. Results (1) After treatment, The levels of ET-1 in EBC and serum of the Qingfeitang treatment group were significantly lower than those of the control group. (2) After treatment, the levels of PGE2 in serum of the Qingfeitang treatment group were significantly lower than that of the control group. (3) The oxygenation index difference before and after treatment of Qingfeitang in the treatment group was higher than in the control group. (4) The duration of mechanical ventilation of the Qingfeitang treatment group was significantly less than that of the control group. Conclusions Qingfeitang could be an effective method for alleviating acute respiratory distress syndrome.

OBJECTIVETo observe the clinical significance of nitric oxide (NO) and 8-isoprostane (8-isoPG) changes in exhaled breath condensate ( EBC) of acute respiratory distress syndrome (ARDS) patients after treated by Qingfei Decoction (QD).

METHODSTotally 48 ARDS patients receiving mechanical ventilation were equally assigned to the QD treatment group and the control group by random digit table. EBC specimens were collected by modified Ecoscreen breath condensate collector (German JAEGER Company) on the first day and the fifth day after confirmed diagnosis of ARDS. Concentrations of NO and 8-isoPG in EBC were measured by ELISA. The oxygenation index and APACHE II scores were recorded at the same time.

RESULTS(1) The fatality rate in the QD treatment group was lower than that in the control group (8.3% vs 37.5%, P < 0.05). (2) After treatment NO and 8-isoPG concentrations in EBC were lower in the QD treatment group (34.49 ± 5.67 µmol/L, 30.09 ± 7.89 ng/L) than in the control group (39.78 ± 9.27 µmol/L, 35.65 ± 8.90 ng/L; P < 0.05). (3) After treatment improved oxygenation index value was higher in the QD treatment group than in the control group (120.88 ± 35.16 vs 101.50 ± 37.70, P < 0.05). After treatment APACHEII scores was lower in the QD treatment group than in the control group (6.21 ± 3.51 vs 10. 26 ± 4.33, P < 0.05).

CONCLUSIONTreatment of ARDS patients by QD was favorable in controlling inflammation, alleviating lung injury, and improving clinical efficacy.

Breath Tests ; Dinoprost ; analogs & derivatives ; analysis ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Inflammation ; Nitric Oxide ; analysis ; Respiration, Artificial ; Respiratory Distress Syndrome, Adult ; drug therapy

OBJECTIVETo examine the effects of H-ras gene silence on cell cycle, proliferation and apoptosis of salivary adenoid cystic carcinoma -M (SACC-M) cell lines.

METHODSThe plasmid H-ras-shRNA, containing the sequence of shRNA targeting H-ras, and HK-shRNA (without interfering effect) were constructed and transfected into SACC-M cells. The cell line with shRNA plasmid stable expression was isolated by G418. The expression levels of H-ras were detected by RT-PCR and protein immunofluorescent assay; cell cycle and cell apoptosis were analyzed by flow cytometry (FCM). The proliferation of cell was also determined by subcutaneous tumor formation in nude mice.

RESULTSAfter transfection of H-ras-shRNA plasmid, the mRNA expression of H-ras in SACC-M cells was down-regulated by 61.80% and protein expression of H-ras was inhibited by 62.76%; the cell proliferation was inhibited obviously; the G0G1 phase cells were increased. The cell apoptosis rate of H-ras-shRNA group was significantly higher than that of HK-shRNA group (P <0.05). The volume of subcutaneous tumor in nude mice was significantly smaller in Hras-shRNA group than in control group.

CONCLUSIONSThe recombinant plasmid HRAS-shRNA could efficiently down-regulate the expression of H-ras gene and protein, induce apoptosis of SACC-M cells and simultaneously inhibit proliferation of these cells in vitro and in vivo.

Animals ; Apoptosis ; Carcinoma, Adenoid Cystic ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Gene Silencing ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins p21(ras) ; genetics ; RNA, Small Interfering ; genetics ; Salivary Gland Neoplasms ; genetics ; pathology ; Transfection

OBJECTIVETo investigate the relationship between the resuscitation promoting role of resuscitation promoting factor and the initial bacteria amount of dormant Mycobacterium tuberculosis.

METHODSMycobacterium tuberculosis (dormant bacteria) was cultured for 100 days, then diluted into 1 mg/ml concentration with 7H9, and further diluted into 0.5, 0.25, 0.125, 0.0625, and 0.03125 mg/ml. Twelve new tubes added with 5 ml 7H9 and divided into two groups: the first group was added with the resuscitation-promoting factor protein, and the second group as control was added with 7H9. In each group the above diluted solutions were added. The tubes were located at 37 degrees C for culture. Optical density (OD) was detected on day 15, 25, 30, and 35. From each tube 1 microl culture solution was plated on 7H11 medium for colony counting.

RESULTSOD detection showed that bacteria proliferation in each group had positive linear correlation (P < 0.05, P < 0.01), indicating that the resuscitation-promoting factor played a similiar role in solutions with different dilution concentrations. 7H11 results and the OD results show that these two detection methods in each group had linear correlation (P < 0.05, P < 0.01), indicating that these two methods showed consistent test results.

CONCLUSIONThe resuscitation-promoting factor has no effect on the resuscitation of dormant Mycobacterium tuberculosis and its initial bacteria amount.

Bacterial Proteins ; metabolism ; Cytokines ; metabolism ; Mycobacterium tuberculosis ; physiology ; Resuscitation

OBJECTIVETo establish a rapid, inexpensive, and simple drug susceptibility test (DST) for Mycobacterium tuberculosis (M. tb) and evaluate its feasibility.

METHODWe used nitrate reductase combined with mycobacteriophage assay (PhaB-NRA) to test 49 clinical M. tb isolates of, and the results were compared with those of PhaB-NRA and traditional absolute concentration method.

RESULTSThe sensitivity, specificity, and accuracy of PhaB-NRA for rifampicin were 89.1%, 91.67%, and 89.8%; on the contrary, those of isonicotinyl hydrazide were 86.21%, 90.0%, and 87.8%, respectively. The coincidence between PhaB-NRA and traditional assay were 0.746 for rifampicin and 0.750 for isonicotinyl hydrazide.

CONCLUSIONSPhaB-NRA is an inexpensive, rapid, and simple DST method. It is a promising rapid screening technique for DST of M. tb.

Anti-Bacterial Agents ; pharmacology ; Biological Assay ; methods ; Humans ; Microbial Sensitivity Tests ; methods ; Mycobacteriophages ; physiology ; Mycobacterium tuberculosis ; drug effects ; Nitrate Reductase ; metabolism ; Rifampin ; pharmacology ; Sensitivity and Specificity

OBJECTIVETo investigate the heparitinase (HPA) expression and cell invasiveness in human ovarian cancer cell line SKOV3 during hypoxia, and explore their relationship with hypoxia inducible factor-1alpha (HIF-1alpha).

METHODSSKOV3 cells were incubated with normoxia, hypoxia, and hypoxia plus rapamycin, respectively. SKOV3 cells of hypoxia group were incubated in 5% CO2 + 1% O2. Cells in hypoxia plus rapamycin group were incubated with 10 ng/ml of rapamycin before cultured under hypoxic condition. Cells in each group were collected for analysis after incubated with hypoxia for 12, 24, and 36 hours, respectively. Western blotting and RT-PCR were performed to detect the expressions of HIF-1alpha and HPA. Cell invasiveness was measured by matrigel invasion assay.

RESULTSWestern blotting showed that the expression of HIF-1alpha significantly increased compared with normoxic group (P < 0.05). However, hypoxia had no obvious impact on HIF-1alpha mRNA expression. The expressions of HPA protein and mRNA of SKOV3 cells of hypoxia group were significantly higher than normoxic group (P < 0.05). The up-regulation of HPA expression in hypoxic group decreased after the utilization of rapamycin. The cell invasion of hypoxic group was significantly higher than that of normoxic group (P < 0.05); meanwhile, the expression of HPA was positively correlated with the invasiveness of SKOV3 cells (r = 0.9863, P < 0.05).

CONCLUSIONHypoxia may promote the expression of HPA and the invasiveness of SKOV3 cells through the HIF-1alpha pathway, which plays an important role in the pathogenesis of ovarian cancer.

Cell Hypoxia ; Cell Line, Tumor ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Neoplasm Invasiveness ; Ovarian Neoplasms ; enzymology ; genetics ; metabolism ; pathology ; Polysaccharide-Lyases ; genetics ; metabolism ; Signal Transduction ; Up-Regulation

BACKGROUNDThe precise mechanisms responsible for the development and growth of intracranial arteriovenous malformations (AVMs) remain unclear. Osteopontin (OPN) is a phosphorylated glycoprotein with diverse functions. This study aimed to analyze the expression of OPN in human brain AVMs.

METHODSThe AVM nidus was surgically obtained from patients with AVM, whereas control brain artery specimens were surgically obtained from patients with epilepsy. Reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the expression of OPN mRNA in biopsy specimens. OPN protein expression was localized by immunohistochemistry. The statistical differences between different groups were assessed by two-way analysis of variance (ANOVA).

RESULTSWe analyzed 36 brain AVM specimens and 8 control brain artery specimens. Eleven patients with brain AVM received embolization treatment, and five underwent gamma knife radiotherapy before resection. Nineteen patients with brain AVM had a history of hemorrhage from AVMs. The expression of OPN mRNA was significantly higher in AVMs than that in the control specimens (25.76 ± 2.71 vs. 21.46 ± 2.01, P < 0.01). There was no statistically significant difference in the extent of OPN mRNA expression between the AVM group with and that without history of hemorrhage (26.13 ± 2.45 vs. 25.34 ± 2.99) or gamma knife radiotherapy (24.39 ± 2.10 vs. 24.53 ± 1.85). However, the difference between the AVM group with and that without embolization treatment history was statistically significant (24.39 ± 2.10 vs. 28.80 ± 1.13, P < 0.01). In the group with gamma knife radiotherapy history, OPN expression was found in arteries with early-stage radio-effect.

CONCLUSIONSOPN may contribute to the vascular instability of brain AVMs. It may play an important role in the pathophysiological process related to embolization treatment.

Analysis of Variance ; Brain ; metabolism ; pathology ; Immunohistochemistry ; Intracranial Arteriovenous Malformations ; genetics ; metabolism ; pathology ; Osteopontin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo determine the correlation between methylation of p16 gene in promoter region and the carcinogenesis and progression of squamous cell carcinoma (SCC) of buccal mucosa.

METHODSMethylation of pl6 gene in SCC and leukoplakia of buccal mucosa was investigated by MSP and pl6 protein was analyzed by Western blot.

RESULTSThe methylation of p16 gene was found in 15 of 30 cases SCC and 1 of 10 cases of leukoplakia of buccal mucosa (P < 0.05). Methylation of p16 gene in SCC of buccal mucosa was not related with age, sex, cell differentiation and clinical stage. But methylation of p16 in the cases with lymph node-metastasis was higher than that in the cases without lymph node-metastasis protein (P < 0.05). Meanwhile Methylation of p16 gene was positively correlated with no-expression of p16 protein (P < 0.01).

CONCLUSIONSThe methylation of p16 gene leaded to the inactivation of p16 gene and was related with the carcinogenesis and progress of SCC of buccal mucosa.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cheek ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; Genes, p16 ; Humans ; Leukoplakia, Oral ; embryology ; genetics ; pathology ; Mouth Neoplasms ; genetics ; metabolism ; pathology ; Promoter Regions, Genetic