Objective: To isolate Mycobacterium tuberculosis H37Rv and H37Ra genes especially,and to construct two cDNA libraries by using suppression subtractive hybridization (SSH).Methods: We used suppression subtractive hybridization (SSH) to clone the differential genomic genes between Mycobacterium tuberculosis virulence strain H37Rv and attenuated strain H37Ra.After two times of subtractive hybridization and two times of PCR,the products of last PCR amplification were inserted into pGEM-T Easy vector and be transformed into the E.coli DH5α and screened of blue and white clones of the transformants.The subtracted cDNA library of differentially expressed genes were identified by RT-PCR.Results:High or especially expressed genes as tester had been obtained by SSH in correctitude reaction (H37Rv as tester) and reverse reaction (H37Ra as tester),the cDNA libraries of A and B of Mycobacterium tuberculosis H37Rv and H37Ra were successfully constructed.90% of the colonies were white clones,the single band of the colonies was 75% and 80%.Conchision:The cDNA libraries of Mycobacterium tuberculosis H37Rv and H37Ra were successfully constructed using SSH technique,which lay a solid foundation for screening and cloning new specific differentially expressed genes in them.

Objective To investigate the clinical therapeutic effectiveness of fingering expansion anus with the application of acupoint treatment to patients with severe craniocerebral injury. Methods Ninety-one patients with constipation in our hospital from May 2013 - May 2015 were randomized into the control group ( n = 45 ) and the observation group ( n = 46 ) . All patients were implemented with routine care of constipation . Based on those regular treatments , the patients in the observation group received the treatment of anus expansion with fingers accompanying plus acupoint treatment with rhubarb powder for three days. The short-term therapeutic effectiveness was evaluated after 3 days of treatment while the long-term therapeutic effectiveness was analyzed 2 weeks later. Results After performing this treatment for 3 days and 2 weeks, the overall clinical symptoms of constipation in the treatment group was significantly lower than those of in the control group (P<0.05). Both the long-term therapeutic effectiveness after treatment two weeks and the short-term therapeutic effectiveness of the observation group were significantly better than those of the control group (P<0.05). Conclusions Anus expansion anus with fingers together with acupoint application in treating constipation is convenient and accessible. Its high effectiveness and limited side-effects can be the best evldence for its unlimited potential of clinical popularization and application.

Background and purpose: Systemic reports about the prognosis of patients with renal cell carcinoma (RCC) of stage T_(1-3a)N_0M_0 combined with inflammatory enlargement of hilar lymph nodes have not been observed in the literature. This study was to investigate the prognostic role of inflammatory enlargement of hilar lymph nodes in the patients with RCC of stage T_(1-3a)N_0M_0 and its association with clinical features. Methods; Forty-nine patients with RCC of stage T_(1-3a)N_0M_0 combined with inflammatory enlargement of hilar lymph nodes were reviewed and all these patients underwent radical nephrectomy from January 1995 to January 2000. Results: The duration of follow-up was 8-14 years with the average of 7.5 years. Seven patients were lost to follow-up. Seventeen patients without RCC and four with RCC metastases are alive at present. Eighteen patients died of RCC relatad complications and three died of cerebro-cardiovascular disease. Metastases occurred in twenty-two patients one year after surgery. The 5- and 10- year survival rates were 53.1% and 42.9%, respectively. Conclusion: Most of the enlarged hilar lymph nodes in RCC were diagnosed as lymphadenitis. The rate of inflammatory enlargement of hilar lymph nodes in the long-term survival patients with RCC of stage T_(1-3a)N_0M_0 were comparatively low. Regional lymphadenectomy or resection of enlarged inflammatory lymph nodes could not protect the patients from metastasis completely. It is advised that limited lymphadenectomy with resection of enlarged inflammatory lymph nodes should be performed for the patients in RCC combined only with inflammatory enlargement of hilar lymph nodes.

Objective To study the effect of continuous subcutaneous insulin injection(CSII) in treatment of type two diabetic patients with pulmonary infections. Methods From January 2001 to December 2005,83 type two diabetic patients with pulmonary infections were randomized to group CSII (42 cases) and group MSII(41 cases). The period of normalization of the blood glucose and cure of the pulmonary infections, cost and morbidity of low blood glucose were observed. Results Blood glucose of all patients in two groups reduced significantly. Significant difference was observed in the period of cure of the pulmonary infections, usage amount of insulin and morbidity of low blood glucose. In group CSII HbA1c, CRP and CHOL reduced significantly. Fasting CRP and high density lipoprotein-C (HDL-C) were remarkably increased. Conclusions Consecutive subcutaneous insulin pump injection is able to control the blood glucose better, to correct metabolic disorder, reduce the cost and hospital day and decrease the morbidity of low blood glucose. CSII is a better of method.

Objective To evaluate the efficacy and safety of sorafenib in the treatment of Chi-nese patients with metastatic renal cell carcinoma. Methods This muhicenter phase Ⅱ clinical trial was performed from May 2006 to December 2006. Sixty-two patients with metastatic renal cell carci-noma not suitable for curative treatment were enrolled. All patients received oral sorafenib as single a-gent at the dose of 400 mg twice a day until disease progression or intolerable toxicities occurred. Re-salts Partial responses were recorded as best response in 11 patients, while complete remission was found in 1 patient and stable diseases were found in another 35 patients. According to the intents-to-treatment population, the overall response rate was 19.4% (12/62), and the disease control rate was 77.4%(48/62). The median progression free survival time was 9.6 months with 1-year progression-free survival rate of 41.9%. However, the median survival time had not reached due to the short fol-low-up. The most frequent adverse events included alopecia (66.1%), diarrhea (62.9%), hand-foot syndrome (58.1%), anorexia (40.3%), rash(37.1%), fatigue (37.1%), hypertension (35.5%), hoarseness(32.3%), joint pain (25.8%), hypophosphatemia (21.0%), fever (19.4%), nausea (19.4%), abnormal transeaminase( 11.3% ), elevated total bilirubicin( 16.1% ), leucopenia( 12.9% ), bleeding under nail(16.1%), and gum bleeding(11.3%). Grade 3 adverse events included hand-foot syndrome (16.1%), hypertension (12.9%), diarrhea(6.5%), hypophosphatemia (4.8%), joint pain (3.2%), and leucopenia(3.2%). Conclusions Sorafenib has prominent anti-tumor activity in Chi-nese metastatic renal cell cancer patients with most adverse events being grade 1 or 2. More attention should be paid to hypertension and cardio-cerebral vascular events during the application of sorafenib.

Objective To evaluate the effectiveness and safety of terazosin in the treatment of Chinese benign prostatic hyperplasia (BPH) patients. Methods A multicenter prospective postmarketing observational study was conducted from June 2007 to March 2008 in 32 urologic centers.Patients were given terazosin for 4 weeks according to the routine medical care procedures following instructions. Effectiveness evaluation included the primary endpoint focusing on the changes in IPSS total score at the end of 2nd and 4th week compared with the baseline. The secondary endpoints were the changes in Qmax and QOL at the end of 4th week, diastolic and systolic blood pressures at the end of 2nd and 4th week compared with the baseline and the discontinuation rate of terazosin within the four weeks. Safety was assessed by adverse events. Results There were 1006 patients included in this study (FAS) and 992 patients (PP) completed the study. Among them, there were 344 patients having hypertension. The total IPSS score reduced from 22.32±6. 13 at baseline to 16. 98±5.92 at the end of the 2nd week and to 14.00±5. 52 at the end of the 4th week in FAS population (P＜0. 01).The total IPSS score changed from 22.32±6.15 at baseline to 16. 96±5.93 at the end of the 2nd week and to 13. 95±5.52 at the end of the 4th week in the PP population (P＜0.01). The efficacy rate was 26.54% at the 2-week treatment and 60.64% at the 4-week treatment, which was defined as obtaining improvement by 30% compared with the baseline. Patient's IPSS in different age groups with different prostatic hyperplasia levels and patients combined with or without 5-α reductase inhibitors were all decreased significantly(P＜0.01). With 4-week treatment of terazosin, Qmax and QOL were improved significantly by 32% and 45% (P＜0.01). Terazosin decreased BPH patient blood pressure with untreated or uncontrolled hypertension (P＜0.05), but had little influence on normal blood pressure of those under control. The incidence of adverse reactions was low. The most common adverse event was dizziness (3.68%). At the end of the study, 960 subjects (95%) were taking drug continuously.Conclasions Terazosin can significantly improve the symptoms and quality of life in Chinese BPH patients with good safety and compliance.

Objective To investigate the correlation between the distnioution of cerebral atherosclerotic stenosis and early neurologic deterioration (END) in patients with acute large artery atherosclerotic stroke.Methods Patients with acute large artery atherosclerotic stroke admitted to the Department of Neurology,the Second Affiliated Hospital of Anhui Medical University from March 2017 to May 2018 were enrolled retrospectively.END was defined as the National Institutes of Health Stroke Scale (NIHSS) score increased by 2 from the baseline within 72 h of admission,or the NIHSS consciousness level score increased by 1,or the NIHSS motor score increased by 1,or having any new neurological deficit.According to whether the patients had END or not,they were divided into END group and non-END group.According to cerebral artery stenosis (stenosis degree ＞ 50％) identified by head and neck CT angiography,magnetic resonance angiography or digital subtraction angiography,they were divided into single artery stenosis group and multiple artery stenosis group.Multivariable logistic regression analysis was used to analyze the independent risk factors for END.Results A total of 371 patients were enrolled in the study,of which 92 (24.8％) had END.In the single artery stenosis group,the incidence of END varies with the distribution of vessel stenosis:anterior cerebral artery (2.3％),middle cerebral artery (54.4％),posterior cerebral artery (9.1％),basilar artery (4.5％),intracranial internal carotid artery (11.4％),intracranial vertebral artery (6.8％),extracranial internal carotid artery (6.8％),and extracranial vertebral artery (4.5％).The incidence of middle cerebral artery stenosis was significantly higher in the END group than that in the non-END group (54.5％ vs.21.2％;x2=17.615,P ＜ 0.001).In the multiple artery stenosis group,the incidence of END was the highest in patients with only intracranial stenosis (66.7％),followed by patients with intracranial and extracranial stenosis (29.2％),and patients with only extracranial stenosis (4.2％).The incidence of only intracranial multi-artery stenosis was significantly higher in the END group than that in the non-END group (66.7％ vs.47.6％;x2 =5.262,P =0.022).Multivariate logistic regression analysis showed that middle cerebral artery stenosis (odds ratio,1.805,95％ confidence interval 1.217-2.676;P=0.003) was an independent risk factor for END.Conclusions END was associated with the distribution of cerebral atherosclerotic stenosis in patients with acute large artery atherosclerotic stroke.The middle cerebral artery stenosis was an independent risk factor for END.

Objective:To analyze the relationship between antithrombin Ⅲ(AT-Ⅲ) activity and survival, bleeding and thrombosis complications in patients with acute-on-chronic liver failure (ACLF), and to explore the prediction value of AT-Ⅲ activity in the prognosis of ACLF patients.Methods:The clinical data of 130 hospitalized patients with ACLF were retrospectively collected in Wuxi No.5 People′s Hospital from January 1, 2013 to April 1, 2019. The liver function, international normalized ratio (INR), and 90-day survival rate were detected. The AT-Ⅲ activity values at admission, week two, week four, and week eight of hospitalization were recorded, and the occurrences of fecal occult blood and femoral vein thrombosis were also recorded. The measurement data were compared by t test, analysis of variance, or rank sum test, and the categorical data were compared by chi-square test. The risk factors affecting the survival of ACLF patients were analyzed by Cox regression. The survival analysis was performed using the Kaplan-Meier method. Results:At the end of 90-day follow-up of 130 patients, 56 patients died, 20 patients (15.38%) were fecal occult blood positive and 15 (11.54%) had femoral vein thrombosis. The baseline AT-Ⅲ activity in the death group was lower than that in the survival group ((17.89±13.68)% vs (36.03±11.96)%), and the difference was statistically significant ( t=-8.045, P<0.01). The baseline AT-Ⅲ activities in fecal occult blood positive and negative patients were (18.26±11.52)% and (25.06±10.97)%, respectively, and in femoral vein thrombosis and non-thrombotic patients were (17.55±10.33)% and (32.48±11.88)%, respectively. The differences were both statistically significant ( t=8.746 and 8.090, respectively, both P<0.01). Through dynamic monitoring of AT-Ⅲ, the AT-Ⅲ activity showed a downward trend in the death group, while that showed an upward trend in the survival group, but the differences were not statistically significant ( F=0.282 and 0.401, respectively, both P>0.05). The Cox regression analysis suggested INR (odds ratio ( OR)=1.364, 95% confidence interval ( CI) 1.078-1.726, P=0.010) and AT-Ⅲ activity ( OR=0.930, 95% CI 0.906-0.954, P<0.01) were the independent factors affecting the survival of patients with ACLF. The area under the receiver operator characteristic curve of the AT-Ⅲ activity for predicting 90-day survival outcome of the patient was 0.706 (95% CI 0.773-0.952, P<0.01), and the cut-off value was 25%. Patients with AT-Ⅲ activity ≥ 25% had a higher survival rate than those with AT-Ⅲ activity <25% ( χ2=58.20, P<0.01). Conclusions:AT-Ⅲ activity is associated with fecal occult blood positive and femoral vein thrombosis in ACLF patients. The AT-Ⅲ activity is an independent influencing factor for predicting the prognosis of ACLF patients. Patients with AT-Ⅲ activity less than 25% have the higher mortality rate.

Objective To investigate the risk factors of early neurological deterioration (END) in patients with cerebral infarction induced by symptomatic intracranial artery stenosis.Methods One hundred and eighty-nine cerebral infarction patients with symptomatic intracranial artery stenosis were collected. According to National Institutes of Health Stroke Scale (NIHSS) scores at 72 h of admission minus baseline NIHSS scores, these patients were divided into END group (n=51,≥2) and non-END group (n=138, <2). Clinical data and laboratory results were retrospectively collected. Univariate Logistic regressionwas used to analyze the differences of above data between the two groups, and multivariate Logistic regression was used to analyze the risk factors of END in patients with cerebral infarction induced by symptomatic intracranial artery stenosis.Results There were significant differences between the END group and the non- END group in age ([69.1±10.6] yearsvs.[65.8±10.4] years), baseline NIHSS scores (10.6±4.6vs. 5.1±4.1), intracranial artery stenosis site (anterior circulation, 82.4％％vs. 66.7％), intracranial artery stenosis degree (occlusion, 66.7％vs.44.2％), cholesterol level ([6.7± 1.0] mmol/Lvs. [4.8±0.8] mmol/L) and lymphocyte count ([2.3±2.2]×109/Lvs.[1.5±0.6]×109/L,P< 0.05). Multivariate Logistic regression analysis showed that age (OR=2.411, 95％C1: 1.102-5.273,P= 0.028), intracranial artery occlusion (OR=122, 95％CI: 3.635-4102,P=0.007) and baseline NIHSS scores (OR=2.464, 95％CI: 1.189-5.105,P=0.015) were risk factors for END.Conclusion Patients with symptomatic intracranial artery stenosis induced cerebral infarction, especially those with old age, intracranial artery occlusion or low baseline NIHSS scores, need more attention to avoid END.