Objictive The estrogen_like activities of environmental estrogens mediated by estrogen receptor (ER) may be assayed through measuring the expressed product of the reporter gene in the recombinant yeast. Methods The recombinant yeast YM_ADER_ERE was obtained by transformation of pGAD_hER and pLacZi_2ERE, the yeast expression vector of full_length human ER cDNA and reporter vector controlled by estrogen response element (ERE) which were constructed successfully in the lab, by means of the lithium acetate method. Results The activity of ?_galactosidase, the expressed product of the reporter gene lacZ in the yeast YM_ADER_ERE, increased significantly when added in the natural estrogen 17?_estradiol (E2) compared to DMSO. In the experiment with E2 at the final concentration of 10-15 to 10-7 mol/L, the maximum effect (EAmax) and the median effect concentration (EC50) of E2 in the yeast were 94.31 U and 0.34 nmol/L respectively. Conclusion The expression of the reporter gene in the recombinant yeast was dependent on the estrogen ligand, which met with the design idea and requirement of the assay system for environmental estrogens and indicated preliminarily that the recombinant yeast estrogen system (RYES) was established successfully.

Objective To establish the recombinant yeast estrogen system(RYES),a reporter vector controlled by estro-gen response elements(ERE)in yeast was constructed.Methods pLacZi-1RER was obtained first by inserting an ERE which was synthesized artificially into the yeast reporter vector pLacZi which had already been lined by SalI and the terminal dephos-phorized by CIAP.Then,pLacZi-zERE.was obtained by inserting another ERE into the XhoI site nearby the ERE in pLacZi-1ERE.Results Sequence analysis indicated that the yeast reporter vector controlled by two tandem copies of ERE was con-structed successfully.Conclusion The yeast reporter vector controlled by ERE,pLacZi-2ERE,was constructed successfully,which was the very important groundwork for further establishment of the recombinant yeast estrogen system(RYES).

OBJECTIVE:To investigate the characteristics and regulations of antibacterials-induced adverse drug reactions (ADR) in our hospital,and to provide reference for promoting rational drug use. METHODS:233 patients with antibacterials-in-duced ADR in our hospital from Oct. 2013 to Jun. 2015 were selected and analyzed statistically according to patient's age and gen-der,route of administration,the type of antibacterials,occurrence time,organs/systems involved in ADR,etc. RESULTS:There were a large number of ADR in patients age ≤18 and ≥70 years,accounting for 19.3% and 18.5%;the male was more than the female;the incidence of ADR induced by intravenous route was the highest,accounting for 94.0%. Most of ADR was caused by cephalosporins,accounting for 33.9%;ADR often occurred within 1 d after medication,accounting for 63.9%;lesion of skin and its appendants injury was main ADR,accounting for 39.3%. CONCLUSIONS:Adhere to theclear indications,a detailed inqui-ry about allergy history before drug use,right dose,suitable route of administration and course,and timely ADR disposal can re-duce the damage caused by ADR.

Objecfive To explore the value of endoscopic uhrasonography(EUS)in diagnosis and preoperative assessment of esophageal cancer.Methods A total of 57 patients with esophageal cancer were examined by EUS.The image features,depth of invasion and lymph node metastasis obtained by EUS were analyzed and compared with postoperative outcomes.Results Esophageal cancer was characterized as an irregular hypoeehoic mass with partial or total destruction of normal esophageal wall structure under EUS.Compared with post-operative pathological findings,the consistent rates of EUS diagnosis in depth of invasion,T staging,and N staging were 84.2%(48/57),87.7%(50/57)and 84.2%(48/57),respectively.Conclusion EUS exhibited high diagnostic accuracy in T staging of esophageal cancer,which is valuable in managements and prognosis of esophageal cancer.

Objective To analyze the clinical and ultrasonic features of primary squamous cell carcinoma of the thyroid(PSCCT)and diagnostic value of ultrasonography (US) in PSCCT in order to improve the cognition of PSCCT.Methods The clinical data and ultrasonic characters of 5 patients with PSCCT confirmed by pathology post surgery were retrospectively reviewed.Results The average age of the 5 patients was 62 years (range from 46 to 79).All of 5 patients with PSCCT were presented with a painless neck mass in clinical palpation examination.Of them,2 patients had different degrees of difficulty in breathing and swallowing,2 patients had hoarseness.Four of 5 patients had died after surgery with average survival time of 9.5 months.On US,the size of lesions was large(a maximum diameter of 2.9-5.5 cm),all of 5 lesions appeared as single solid mixed-echogenicity mass including irregular lamellate marked hypoechoic region,with irregular margin and undefined boundary.Of them,2 lesions presented with lobulated,only 1 lesion exhibited microcalcification and another 4 lesions had no calcification,3 lesions exhibited a? sign of breakthrough thyroid envelope.Color Doppler flow imaging showed few blood flow (3/5) and moderate blood flow (2/5),a higher vascular resistance spectrum was detected in 4 lesions (RI 0.72-0.88).Three patients exhibited suspicious cervical lymph nodes.Conclusions The characteristic ultrasonic appearance combined with the clinical features enables a confident preoperative diagnosis of PSCCT.

Objective To determine the correlation between Smad4 mRNA,protein expression,abnormal methylation of CpG islands of Smad4 and colorectal cancer.Methods The expression of Smad4 in 43 cases of colorectal cancer and its adjacent tissue.30 cases of colorectal adenoma and 12 normal colon mucosa were analyzed by RT-PCR,sequencing,semi-quantitative RT-PCR,methylation-specific PCR(MSP)and immunohistochemistry.Results The mRNA expression of Smad4 was detected in 25 cases of colorectal cancer(58.14%),with expression levels at 0.73±0.25,which was significantly lower than those in corresponding adjacent tissues(88.37%,0.95±0.29),eolorectal adenoma(90.63%,1.01±0.37)and normal mucosa(100.00%,1.18 ±0.33)(P＜0.05).The positive rates of Smad4 gone promoter methylation in colorectal caner(60.53%)was significantly higher than that in other tissues(27.03%,25.00%and 16.67%,respectively,P＜0.05).The expression of Smad4 protein was significantly lower in colorectal cancer(44.19%)than that in other tissues(81.40%,87.50%,91.67%,respectively,P＜0.05),which was correlated with tumor invasion and lymph node metastasis.Conclusion Down-regulation of Smad4 expression may be associated with the development,the biological behavior and prognosis of colorectal cancer,and it can be an important biological marker in evaluation of disease progression.

AIM: To determine the contents of ephedrine、laetrile、glycyrrhizic acid and liquiritin in Huagai Powder traditional decoction and granule decoction(Herba Ephedrae,Semen Armeniacae Amarum,Radix et Rhizoma Glycyrrhizae,etc.) by HPLC. METHODS: An Agilent SB-C_(18) column was used for ephedrine,the mobile phase was acetonitrile-0.1% phosphoric acid(4∶96),detection wavelength at 207 nm;An Agilent XDB-C_(18) column was used for laetrile,the mobile phase was methanol-water(23∶77),detection wavelength at 215 nm;An Agilent XDB-C_(18) column was used for glycyrrhizic acid,the mobile phase was methanol-0.2 mol/L ammonium-acid(67∶33∶1),detection wavelength at 250 nm; An Agilent XDB-C_(18) column was used for liquiritin,the mobile phase was acetonitrile-0.5% acetic acid(20∶80),detection wavelength at 276 nm. RESULTS: There were discrepancies between traditional decoction and granules in the contents of four ingredients,the average contents in granules were more than those in the traditional decoction. CONCLUSION: The peaks of the four ingredients are(segregated)(effectively),and the method is simple,conveient,exact and has good reproducibility,the other ingredients do not have interferences for the determination.

Objective To investigate the possibility and efficacy of laparoscopic peritoneal vaginoplasty for the treatment of congenital absence of vagina. Methods A total of 4 patients with congenital absence of vagina underwent laparoscopic peritoneal vaginoplasty. Results Operations were performed successfully in the 4 cases. The mean operative time was 114 min (range: 90-160 min), and the mean postoperative hospital stay was 5.4 d (range: 5-6 d). There was no complication in all patients. Conclusion Laparoscopic peritoneal vaginoplasty is a safe, minimally invasive, and reliable method for the treatment of congenital absence of vagina.

Objective To explore the efficacy of craniotomy and aneurysm clipping in elderly patients with ruptured intracranial aneurysms.Methods Totally 75 elderly patients with ruptured intracranial aneurysms in our hospital who underwent craniotomy and intracranial aneurysm clipping:approaching into head by wing point based on CT image,selecting the appropriate position and angle for clipping the aneurysm.Surgical efficacy and postoperative complications were observed.Prognosis was assessed by Glasgow Outcome Scale (GOS).Results 77 aneurysms presented in 75 patients and 76 aneurysms were clipped.The main postoperative complications were intracranial hematoma (5 cases),pulmonary infection (4 cases),electrolyte imbalance (10 cases),intracranial infection (2 cases),arrhythmia (3 cases),gastrointestinal bleeding (5 cases),cerebral infarction (3 cases).GOS assessment demonstrated that prognosis in 48 cases was good (64.0％),16 cases was moderate disability (21.3％),6 cases was severe disability (8.0％),2 cases was vegetative survival (2.7％),3cases was in death (4.0％).The cure rate was 64.0％ and the overall good-prognosis rate was 85.3％.Conclusions Craniotomy and aneurysm clipping has a better surgical efficacy and a high cure rate in elderly patients with ruptured intracranial aneurysm.

AIM: To explore the role of TGF-?_1 and signaling transduction molecule, Smad4, in the development of glomerulosclerosis. METHODS: Expression levels of TGF-?_1, Smad4, collagen Ⅰ proteins were evaluated by immunohistochemistry in renal biopsies from 38 cases with a spectrum of glomerulonephritis, and compared with 20 normal kidney tissue with image analysis system. After stimulation with TGF-?_1, expressions of endogenous Smad4 and collagen Ⅰ mRNA and proteins and its modulation by TGF?_1 were evaluated by RT-PCR and Western blotting analyses in cultured human mesangial cells. RESULTS: All types of proliferative and sclerotized glomerulonephritis showed an increased expression of TGF-?_1, Smad4 and collagen Ⅰ ompared with the 20 normal kidney tissue in glomerular (P