Changes of syndromes and therapeutic feature of the patients with steroid-dependent asthma(SDA)during steroid withdrawal were investigated.It was held that the patient presented pathological changes from deficiency of Yin and hyperactivity of evil fire before steroid withdrawal to deficiency of Yin and Yang during steroid withdrawal to deficiency of kidney-Yang after steroid withdrawal,with the syndrome of stasis of phlegm and blood throughout the whole process.Therefore,a new scheme for weaning the steroid with the TCM three-step sequential therapy was established according to the changes of syndromes,i,e., nourishing Yin and blood,clearing away excessive heat and eliminating stasis of phlegm and blood before steroid withdrawal; nourishing Yin and blood,inhibiting pathogenic wind,replenishing Qi and Yang,activating blood circulation,resolving phlegm and lowering the adverse rise of Qi to relieve asthma during steroid withdrawal;warming kidney-Yang and adjusting Qi and blood,and regulating functional activity of Qi after steroid withdrawal.Because the three-steep sequential therapy was designed based on the dynamic change of SDA during steroid withdrawal,it conformed with the whole clinical process of the occurrence,development and changes of SDA than any other single way,and reflected the essential characteristics of the pathological changes.

Objective: To discuss the effect of non-contrast-enhanced magnetic resonance angiography in renal artery imaging. Methods:Twenty patients were selected with suspected as the research object, using non-contrast-enhanced MR examination, simultaneously enhanced MR examination, and relatively non-enhanced imaging examination effect on the results for statistical analysis and processing. Results: Non-contrast-enhanced ratio comparing contrast-enhanced, not statistically significant (x2=1.058, P>0.05). Conclusion:Non-contrast MR angiography is an effective, safe, non-invasive, high accuracy of the evaluation of renal artery screening tool, worthy of clinical application.

Objective To investigate the effects of Xuebijing injection on expression of myeloid cell triggering receptor-1(TREM-1)and the plasma levels of soluble TREM-1(sTREM-1),tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)in patients with severe sepsis. Methods Twenty patients with severe sepsis admitted into Critical Care Medicine Department,Affiliated Hospital of Nantong University were given comprehensive treatments according to the guidelines for management of severe sepsis and septic shock(2004),and they were divided into Xuebijing group and control group(each 20 cases). The Xuebijing injection group was given Xuebijing injection 50 mL,3 times daily for 5-7 days followed by regular treatments. The changes in blood TREM-1 mRNA expression and plasma concentrations of sTREM-1,TNF-α,and IL-6 were detected before and after treatments on the 3rd and 5th day,and the above indexes were compared between the two groups. Results Before treatments,there were no significant differences in TREM-1 mRNA expression and levels of sTREM-1,TNF-α,IL-6 between two groups (all P>0.05). The TREM-1 expression and plasma concentrations of sTREM-1,TNF-α,and IL-6 of two groups were declined after treatments compared to their baselines,the degree of decline being more prominent in Xuebijing group〔TREM-1 mRNA 3 days:1.065±0.277 vs. 1.217±0.301,t=-3.267,P=0.047;5 days:0.912±0.239 vs. 1.071±0.254,t=-5.072,P=0.032;sTREM-1(ng/L):146.93±13.76 vs. 176.22±19.46,t=-5.442,P=0.033;TNF-α(ng/L):77.51±11.28 vs. 107.72±13.17,t=-4.355,P=0.032;IL-6(ng/L):288.35±14.59 vs. 323.89± 24.51, t=-3.941,P=0.028〕. Conclusion Early implication of Xuebijing injection is of great significance in patients with severe sepsis,it may reduce the expression level of TREM-1 and serum levels of downstream inflammatory mediators,that is beneficial to the control of inflammatory responses and improvement of systemic inflammatory response syndrome in such patients.

Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.

Objective:To observe the effect of three-stage sequential therapy on the functions of hypothalamic-pituitary-adrenal(HPA)axis and levels of the glucocorticoid receptor in patients with steroid-dependent asthma.Methods:55 patients with SDA were divided randomly into two groups,29 patients were treated with three-stage sequential recipes plus Pulmicort Turbuhaler(400?g/d)in the treatment group,while the other 26 patients with Pulmicort Turbuhaler(800?g/d)in the control group. The therapeutic courses were both 12-14 weeks.The plasma concentrations of ACTH and COR and the levels of PBMC GCR were determined before and after treatment.Results:The plasma concentrations of ACTH and COR and the levels of PBMC GCR in the two groups were all increased remarkably and there were significant difference between the two groups after treatment(P

Objective To investigate the factors affecting the survival and prognosis of patients with stage Ⅲ～ Ⅳgastric cancers.Methods A total of 156 cases of Ⅲ ～ Ⅳ gastric cancer was studied retrospectively from September 1,2006 to December 31,2012.Kaplan-Meier analysis,Log-rank univariate analysis,Cox proportional hazards model analysis were used to analyze survival and prognostic factors.Results Twelve cases were lost,to the end of the follow-up,22 cases were alive.The median survival time was 29.3 months.1-year,3-year,and 5-year overall survival rates were 83.3％,37.8％,and 21.2％,respectively.Univariate analysis showed that tumor size,tumor node metastasis (TNM) staging,curative resection,hyperthermic intraperitoneal perfusion chemotherapy (HIPEC),and postoperative chemotherapy were correlated with prognosis (P ＜0.05 for all).Multivariate analysis showed that TNM staging,curative resection,HIPEC,and postoperative chemotherapy were independent prognostic factors (P ＜ 0.01 for all).Conclusions TNM staging,curative resection,hyperthermic intraperitoneal perfusion chemotherapy and postoperative chemotherapy are the independent factors affecting the prognosis of stage Ⅲ～ Ⅳ gastric cancer after resection.Hyperthermic intraperitoneal perfusion chemotherapy and postoperative chemotherapy can improve their survival.

Objective To develop a rapid method with high sensitivity and specificity to detect 4 mycobacterial species(M. tuberculosis, M. avium, M. intracellulare and M. kansasii) which are the most common opportunistic Mycobacteria in AIDS patients. Methods The sensitivities and specificities of PCR were determined with different primer pairs targeting various mycobacterial genes. Multiplex PCR with combination of 4 primer pairs was used to detect the template mixtures of either 1, 2 or 3 mycobacterial DNA. Sensitivities of multiplex PCR were measured. Results Specific DNA fragments of 4 mycobacterial species mentioned above could be detected by PCR and sensitivities ranged from 1 ? 101 ~ 1 ? 102 cells/mL, while the other 17 mycobacterial strains were all PCR-negative. Multiplex PCR could amplify the corresponding 1, 2 or 3 DNA fragments, depending on the number of template DNA added, and sensitivities of multiplex PCR ranged from 1 ? 102 ~ 1 ? 103 cells/mL. Conclusions Multiplex PCR is a rapid, sensitive and specific method for differentiation and detection of Mycobacteria.

Objective To develop a PCR-RFLP method for the identification of eight mycobacterial species. Methods PCR was performed targeting the gene encoding 65-kDa heat shock protein which was common to all mycobacteria. Two restriction enzymes, BstE Ⅱ and Hae Ⅲ, were used to digest the PCR products, and specific restriction patterns of different mycobacteria were obtained. Results The specific restriction patterns of different mycobacteria were identical to the data previously reported. Conclusion We could differentiate M. avium, M. intracellulare, M. kansasii, M. tuberculosis, M. scrofulaceum, M. marinum, M. fortuitum and M. chelonae in one experiment by PCR-RFLP.

Objective To investigate the effect of hyperthermic intraperitoneal perfusion and cisplatinon apoptosis in human ovarian cancer cells.Methods Two human ovarian cancer cells (OVCAR-3,A2780) were divided into control group,cisplatin group,hyperthermia group and thermo-chemotherapy group;microscopy was used to observe the morphological changes of the four groups;AO/GV stain and flow cytometry(FCM) was used to analyze cell apoptosis;Apoptosis related genes caspase7,caspase8 and Bax in ovarian cancer cells were detected by fluorescence quantitative PCR.Results Inverted microscopy observeed that the ovarian cancer cells retracted and suspended partially in the cisplatin group and hyperthermia group,especially in the thermo-chemotherapy group.After AO/GV staining,the apoptotic cells were increased in the cisplatin group and hyperthermia group compared with the control group,and the thermo-chemotherapy group was more than cisplatin group and hyperthermia group.FCM results indicated that the proportion of cells apoptosis were higher in the cisplatin group and hyperthermiagroup,the thermo-chemotherapy group is the higher than the all other groups(P＜0.05).q-PCR results showed that in the thermo-treatment group the expression of pro-apoptotic genes,including caspase3,caspase6,caspase7,caspase8,caspase9,Bax,Bak and Bid,was significantly higher than other groups,apoptosis inhibitory gennes,such as Bcl-2,Bcl-xL,Mcl-1,c-FLIP,was significantly decreased than the others.Conclusions Cisplatin plus hyperthermia can promote the apoptosis in ovarian cancer cells.

Objective To investigate the efficacy of precise hyperthermic intraperitoneal perfusion chemotherapy (HIPEC) in the treatment of advanced pancreatic cancer.Methods Thirty-six patients with advanced pancreatic cancer were analyzed retrospectively.Eighteen patients who received HIPEC combined with radiotherapy and chemotherapy were assigned as the treatment group and the other 18 patients who received chemotherapy and radiotherapy were assigned as the control group.Recent curative efficacy,Karnofsky Performance Status (KPS) score,postoperative complications and survivals between the two groups were analyzed,respectively.Results Significant differences were found between two groups in total short-term effective rate (P ＜ 0.05).The total short-term effective rate of treatment group and control group were 66.67％ (12/18) and 27.78％ (5/18),respectively.The increment of KPS score of treatment group was significantly higher than that of control group (P ＜ 0.05).There was no significant difference between two groups in postoperative complications (P ＞ 0.05).The median overall survival time (OS) of treatment group was 11 months (7 ～ 31 months),and the median OS of control group was 7 months (4 ～ 18 months).The survival of the treatment group was longer than the control group (P ＜ 0.05).Conclusions HIPEC treatment improved significantly the survival and life quality of advanced pancreatic cancer patients.With acceptable morbidity and mortality rates,HIPEC regime was an effective treatment modality for patients with advanced pancreatic cancer.