Objective To clone human beta-2 adrenoceptor gene from human bladder smooth muscle and to construct its antisense eukaryotic expression vector. Methods The ?_2-AR full length cDNA was cloned from human detrusor cells through RT-PCR and subcloned into clone vector (pUC18).The objective gene was then cut from ClaⅠ/HindⅢ sites of pUC18 with restriction endonulcease and subcloned into pLNCX vector in trans-direction.Finally the constructed ?_2-AR gene antisense expresstion vector was identified through restriction endonuclease analysis and sequencing. Results The sequence of cloned ?_2-AR full length cDNA was certified by comparison with the database of the Genebank.The constructed antisense eukaryotic expression vector was proved to be same with designed by restriction endonuclease analysis and sequencing. Conclusions ?_2-AR full length cDNA was cloned and its antisense eukaryotic expression vector was successfully constructed.This technique establishs the foundation for the further research on drug treatment of bladder dysfunction.

AIM: To establish the method of determining psoralen and isopsoralen in Bushen Yinao Capsules(Fructus Psoraleae,Radix et Rhizoma Ginseng Rubra,Poria,etc.). METHODS: HPLC conditions were as(follows:) Chromasil C_(18) column,a mobile phase of CH_3OH-H_2O(55∶45),and the wavelength at 247 nm. RESULTS: This method showed a good linear relationship in the ranges of 0.038-0.309 ?g(psoralen) and 0.036-0.295 ?g(isopsoralen).The average recoveries were 99.50% for psoralen(RSD was 0.91%,n=5) and 100.52% for isopsoralen(RSD was 1.70%,n=5). CONCLUSION: The method is simple,swift,accurate and practical in quality control of Bushen Yinao Capsules.

Purpose To explore the findings and diagnostic values of apparent diffusion coefficient (ADC) and diffusion weighted image (DWI) of the portal venous tumor thrombus (PVTT) caused by hepatic carcinoma.Materials and Methods Thirty-one patients with hepatic carcinoma (43 lesions) with 63 PVTT in the main branches and trunks diagnosed by clinical and MRI were enrolled. All patients underwent conventional MRI (cMRI) imaging, DWI and ADC imaging, the features of cMRI, DWI and ADC were observed, the relevance of ADC values between the hepatic carcinoma lesion and PVTT were analyzed.Results Among the total 43 lesions, DWI image showed low signal, iso-signal and high signal in 1, 4 and 38 lesions, respectively; and ADC image showed low signal, iso-signal and high signal in 36, 5 and 2 lesions, respectively. In the total 63 PVTT, DWI showed low signal, iso-signal and high signal in 4, 7 and 52 lesions, respectively;while their ADC images showed low signal, iso-signal and high signal in 54, 6 and 3 branches, respectively. There was good consistency for the results of two observers on the findings of ADC of tumor lesions (Kappa=0.8334,P<0.05), and a moderate consistency on that of PVTT (Kappa=0.5215,P<0.05). The average ADC value of tumor lesion and PVTT was (1.127±0.268)×10-3 mm2/s and (1.021±0.363)×10-3 mm2/s, respectively; there was a correlation of the mean values of ADC between tumor lesion and PVTT (r=0.246,P<0.05). Conclusion The features such as low signal and low value on ADC image and high signal on DWI obtain a certain clinical application value for qualitative diagnoses of PVTT.

ObjectiveTo explore the efficacy of sevoflurane or propofol combined with remifentanil during the maintenance of general anesthesia in thyroid gland surgery.MethodsSixty patients underwent thyroid gland surgery were randomly divided into tow groups.Once the larynx mask was intubated, anesthesia was maintained with propofol(the effect site concentration was 2.5 ～3.5mg/L) and remifentanil(the effect site concentration was 4.5 ～5.5μg/L) by TCI technique in group P,but with sevoflurane(2％～4％) and remfentanil(the effect site concentration was 2.5 ～4.0g/L)in group S.The depth of anesthesia was measured by the A-line TM AEP Monitor which expressed as A-Line ARX Index TM(AAI).All patients' SBP,DBP and HR were recorded at eight time points: before induction time(T0) ,after induction but before larynx mask intubation time(T1) ,intubate larynx mask time(T2) ,cut skin time (T3), separate thyroid gland time (T4), cut thyroid gland time (T5), remove larynx mask time (T6) ,leave the operation room time(T7).The emergency time,the conscious of the patients after anesthesia and the side effects were also recorded.ResultsThere were no significant differences between the groups with respect to age,gender,weight,the duration of operation,the emergency time and the conscious of the patients after anesthesia.SBP,DBP,HR of the patients in both groups showed no significant difference at T0,T1 ,T2, T3 (all P ＞ 0.05), but had significant difference at T4,T5,T6, T7 (all P ＜ 0.05).Compare with group P,the incidentce of restlessness, dizziness, drowsiness, rigor and pain was significantly lower in group S(all P ＜0.05).The incidentce of nausea,vomit and aspiration did not appear in both groups.ConclusionBoth groups showed good anesthesia effects and the patients also emerged from anesthesia quickly.But the anesthesia maintained with sevoflurane and remifentanil could bring more stable hemodynamics and lower incidence rate of the side effects.

Objective To investigate the effect of different pore sized hydroxyapatite for promoting bone vascularization in tissue engineering.Methods Male Wistar rats were randomly divided into three groups,named group A,B and C,which were im-planted hydroxyapatite bioceramics compositing 4 μg bone morphogenetic protein with different aperture of 200 -300,350 -450, 500-600 μm in the back subcutaneously.The size of each block was 5 mm×5 mm×1 mm in a weight about of 40.0 mg.After im-plantation,the animals were killed and the implants and the surrounding tissue were taken out at the first,second,third and forth week respectively.HE staining of histological analysis was used to detect the situation of local neovascularization.Results There was significant difference between second and third week in group A.Comparing the area of vascularization at different time points in group B and group C,there were significant difference in the comparison of intragroup (P <0.05 ).During the first week after surgery,there was only group C that had the area of vascularization.During the second and forth week after operation,the area of vascularization in group B and group C were significant higher than group A (P <0.05).The C group showed a great deal of new-born blood vessels and clear formation of bone trabeculae.Conclusion The hydroxyapatite bioceramics of 500-600 μm could better promote vascalarization of tissue engineering in bone.

Objective To evaluate the effect of age on the median-effective target plasma concentration (EC50) of propofol inhibiting body movement evoked by gastroscopy in the patients.Methods Ninety adult patients of both sexes,of ASA physical status Ⅰ or Ⅱ,with body mass index 19-25 kg/m2,scheduled for elective gastroscopy,were divided into 3 groups according to age (n =30 each):18-39 yr group (Ⅰ group),40-64 yr group (Ⅱ group) and 65-85 yr group (Ⅲ group).In Ⅰ,Ⅱ,Ⅲ groups,propofol was given by target-controlled infusion with the initial target concentrations of 2.5,2.0 and 1.5 μg/ml,respectively,and gastroscopy was performed when the target concentration was achieved.Body movement was defined as the directional movement in head or four extremities during gastroscopy.The target plasma concentration of propofol was determined by up-and-down sequential trial.Each time the plasma concentration of propofol increased/decreased by 0.5 μg/ml in the next patient depending on whether or not body movement developed.The EC50 and 95 ％ confidence interval of propofol inhibiting gastroscopy-evoked body movement were determined using Probit analysis.Results The EC50 (95 ％ confidence interval) of propofol was 4.2(3.8-4.5),4.1(3.7-4.4) and 2.4(1.8-2.7) μg/ml in Ⅰ,Ⅱ and Ⅲ groups,respectively.There was no significant difference in the EC50 of propofol between group Ⅱ and group Ⅰ.The EC50 of propofol was significantly lower in group Ⅲ than in Ⅰ and Ⅱ groups.Conclusion Age affects propofol-induced analgesia in patients with visceral pain,and the potency of propofol inhibiting visceral pain during gastroscopy in the elderly patients is significantly enhanced as compared with that in the young and middle-aged patients.

Objective To study the effects of extracted active components of Chaenomeles Speciosa (EACCS) on non-alcoholic fatty liver disease (NAFLD) in mice; To discuss the possible molecular mechanism. Methods Forty male KM mice were randomized into four groups, namely normal group, model group, low-dose (50 mg/kg) EACCS group and high-dose (100 mg/kg) EACCS group. Except that the normal group was daily given routine diet, the other groups were given high-fat–high-fructose diet (HFFD). The mice were put to death 4 weeks later. Body weight, liver weight and serum TG were measured. HE and oil red O staining were used to observe liver tissue morphology. RT-PCR and Western blot were used to detect the expression of lipid metabolism related genes. Results Compared with the normal group, the liver size, liver index (P<0.01) and epididymal fat index (P<0.05) increased significantly;The ALT and GLU in serum increased (P<0.05), TG increased (P<0.05), and pathological findings showed significant steatosis; RT-PCR and Western blot showed that the expression levels of SIRT1 and FoxO1 mRNA decreased and the level of SERBP-1c increased in the model group. Compared with the model group, the hepatic lipid accumulation of EACCS groups was obviously improved, and the serum ALT, GLU, and TG levels significantly decreased, the expression levels of hepatic SIRT1 and FoxO1 mRNA increased. Conclusion EACCS has protective effects on NAFLD mice induced by HFFD, and its mechanism may be related to the activation of SIRT1-FoxO1 signaling pathway in the liver tissues.

Aim: To investigate the pathological changes in NTBC[2-(2-nitro-4-trifluoro-methyl-benzoyl) -1,3 cy-clohexanedione]-induced hepatic injury in mice and in the repopulation of adult hepatocytes in Fah~(-/-) mouse. Methods: Autogenous hepatic injuries in Fah~(-/-) mice were induced by the treatment of NTBC. Injection of hepatocytes obtained from wild-type mice to spleen were transplanted into the Fah~(-/-) mice. Then, changes to body weight and the likelihood of the transplanted Fah~(-/-) mice, and hepatic immunohistochemistry were ob-served. In addition, pathological changes to liver damage induced by NTBC treatment were analyzed under HE-staining microscopy and electron microscopy. Results: The surviving Fah~(-/-) mice subjected to hepatocyte trans-plantation were found to be healthy and in stable body weight. liver repopulation reached to 90% in the 8th week. Repopulating hepatocytes caused no alteration to histological structure of the recipient liver, and subacute hepatic injury occurred in the Fah~(-/-) mice after NTBC treatment. Electronic microscopy observations indicated that necrosis in the hepatocytes occurred at early stage and that apoptosis gradually appeared. It was also shown both necrosis and apoptosis co-existed in the same samples of interest at the following stages of the induced liver injury. Conclusion: Transplanted hepatocytes proliferated in Fah~(-/-) mice allow 90% of the hepatocytic repopula- tion. Repopulation renders normal hepatic function and structure in the recipient Fah~(-/-) mice, as a model of liver repopulation, could be applicable in study of stem cell derived hepatic cells in transplantation assay.

Objective:To explore the clinical characteristics, diagnosis and treatment of acute leukemia patients during pregnancy.Methods:The clinical data of 16 cases with acute leukemia during pregnancy from January 2009 to December 2018 in the First Affiliated Hospital of USTC were retrospectively analyzed. The diagnosis and treatment regimens, pregnancy outcome, the early fetus and survival status of patients were also analyzed.Results:All 16 leukemia cases were confirmedly diagnosed and classified by bone marrow puncture, including 13 cases of acute myeloid leukemia (5 cases of non-acute promyelocytic leukemia and 8 cases of acute promyelocytic leukemia) and 3 cases of acute lymphoblastic leukemia. At the time of confirmed diagnosis, 6 patients were in first trimester, 6 cases in second trimester and 4 cases in late trimester. As for pregnancy outcome, 1 patient had natural birthing, 5 patients underwent cesarean operation, 9 patients underwent artificial abortion and 1 patient had spontaneous abortion. Chemotherapy was performed in 15 patients during pregnancy, 11 patients received chemotherapy for treatment of primary disease after pregnancy, 3 patients died during the treatment. During the follow-up of 13 cases, 8 patients survived and 5 patients lost follow-up.Conclusions:Early diagnosis of acute leukemia during pregnancy is very important. Bone marrow puncture should be performed timely to make clear diagnosis when blood routine result is abnormal during antenatal care. Multidisciplinary consultation should be initiated in time, and the best treatment plan should be worked out to guard against serious complications during pregnancy.

Objective To investigate the place reaction of support drainage tube in cholecystojejunostomy and its pathological changes.Methods Eight pigs were randomized into 2 groups.The tube-placing cholecystojeunostomy(n=2)or choledochocholedochostomy(n=2)and non-tube-placing cholecystojejunostomy(n=2)or choledochocholedochostomy(n=2)were performed respectively.All the 8 pigs were sarcrificed 3 months after the operation for pathological examination.Results Gross anatomy catheter group showed anastomotic inflammatory hyperplasia, wall hardening hyperplasia,luminal narrow contracture and stone formation.Microscopic biopsy showed bile duct mucosa in interstitial lymphocytes, plasma cells and other chronic inflammatory cell infiltration, a large number of fibrous tissue hyperplasia.However, these features were not found in pigs without placing the tube.Conclusion The tube can not prevent the stenosis.It might promote the formation of stricture since it can stimulate inflammatory hyperplasia and tissue hardening.