BACKGROUND: Wnt signaling pathway plays an important regulative role in the embryonic development processes. Accordingly, it is of great significance to establish the cell model of Wnt signaling pathway so as to conduct study on it. OBJECTIVE: To establish Wnt signaling pathway cell model by transfecting L-M (TK-) cells with Wnt3a eukaryotic expression plasmid, and to investigate the effect of canonical Wnt signal pathway on the β-catenin subcellular distribution. METHODS: The eukaryotic expression plasmid pgk-Wnt3a-pcDNA3.0 after amplification was digested by restriction endonuclease first. Then it was transfected together with the control plasmid pgk-neo-pcDNA3.0 into L-M (TK-) cells via lipofection, after which the cell colony was screened by G418 for amplification. RT-PCR was used for detecting the expression products and the indirect immunofluorescence assay for observing the effect of Wnt3a on the β-catenin subcellular localization of L-M (TK-) cells. RESULTS AND CONCLUSION: The Wnt3a plasmid was verified by endonuclease digestion to have produced the expected plasmids after amplification. According to the RT-PCR detection to the 10 stably-transfected cell colonies achieved by 3 weeks of G418 screening, it was seen, on the L-Wnt3a cDNA, a strip of bright band of 320 bp in length, which showed that the products of amplification were exactly the expected fragments and that the Wnt3a plasmid was expressed on mRNA transcriptional level after being transfected with L-M (TK-) cells. In contrast, no expected band was found on the cDNA of L-M (TK-) calls transfecting the control plasmid. In addition, the immunofiuorescence assay detection showed that the protein expression of Wnt3a was found in the cytoplasm of the L-M(TK-) cells tranfecting Wnt3a plasmid, while for those transfecting the control plasmid, it was opposite. β-catenin, as showing by bright red fluorescence, was found to concentrate and enter into the nucleus of the L-M (TK-) cells transfecting Wnt3a plasmid, while for those transfecting the control plasmid, it was opposite. Cell model with continually activated Wnt signaling pathway is established. The stable expression of Wnt3a in L-M (TK-) cells transfected with pgk-Wnt3a-pcDNA3.0 is obtained. The expression of Wnt3a is able to promote the transfer of β-catenin from cytoplasms into nucleus in L-M (TK-) cells.

It is an important safeguard to realize TCM renovation,raise medical levels and fulfill TCM modernization by elevating information diathesis of TCM personnel.This paper introduced the contents of information diathesis,discussed the current situation of TCM person information diathesis and the importance of elevating their information diathesis.By different kinds of training,how to raise the information awareness,information acquisition ability and information morality of TCM information personnel were further expounded.

Two recombinant plasmids pVAX/Sj23 and pVAX/mIL-18 containing Schistosoma japonicum 23 000 membrane protein (Sj23) and murine IL-18 were evaluated for their ability to induce immune responses and to protect against S. japonicum challenge in mice. All animals vaccinated with pVAX/Sj23 alone or plus pVAX/mIL-18 developed specific anti-SWAP (soluble worm antigen preparation) ELISA antibody and splenocyte proliferation response,and co-injection of pVAX/mIL-18 significantly increased the production of IFN-γ and IL-2 compared with pVAX/Sj23 alone, indicating that IL-18 enhances the Th1-dominant immune response. The challenge experiment showed that worm reduction rates in pVAX/Sj23 group compared with control group (pVAX1) was 26.5% and in the pVAX/Sj23 plus pVAX/mIL-18 group was 41.9% ,and the hepatic egg reduction rates were 42.7 and 49.6%,respectively. These results indicated that co-injection of an IL-18 plasmid with Sj23 DNA vaccine efficiently improves the protective effect against S. japonicum infection.

Objective To evaluate the efficacy of self-retaining suture (QuillTM SRS) in retroperitoneal laparoscopic partial nephrectomy for complicated renal tumor by assessing perioperative parameters.Methods Between 2010 and 2012,78 cases of complicated renal tumor (R.E.N.A.L score ≥ 7) treated by retroperitoneal laparoscopic partial nephrectomy (LPN) with two layers continuous knotless barbed suture (QuillTM SRS group) (n=30) or traditional absorbable vicyl suture (non-SRS group) (n=48) were retrospectively analyzed.In QuillTM SRS group,2-0 Quill SRS was used to suture the deep wound bed,and the second outcr layer renorrhaphy was performed with a 1-0 Quill SRS by the same way.In non-SRS group,the inner layer was sutured using a 15cm in length 2-0 monicryl suture by the same method mentioned above.A second outer layer was sutured with 1-0 vicryl suture across the wound.Cases were matched for R.E.N.A.L score.Comparison was made in term of operation time,preoperative parameter and perioperative complications between SRS group and non-SRS group.Results Renorrhaphy was successfully performed in all cases except 1 case converting to open surgery in non-SRS group.Mean warm ischemia time in SRS group was shorter than non-SRS group (18 vs 25 min,P =0.021).The proportion of bleeding requiring intervention in the non-SRS group (7/48,14.5％) was 4.3-fold higher than that of the SRS group (1/30,3.3％),but the differernce is not significant (P＞0.05).There were no significant differences between two groups in postoperative creatinine changes.Limitations of this study include the absence of randomization and the relative small sample size.Conclusions SRS can be safely used for complicated renal tumor during LPN,and SRS can significantly reduce the WIT and may also reduce bleeding during the operation.

ObjectiveTo report the results of preventive control program of severe thalassemias in Zhuhai City of Guangdong Province from 1998 to 2010.MethodsAs the guide centre of marriage and childbearing and the greatest maternity hospital in Zhuhai City of Guangdong Province,Zhuhai Municipal Maternity and Child Healthcare Hospital constructed the genetic screening network for thalassemias testing and referred for follow-up and for genetic counseling.The couples for premarital medical examination or regular healthcare examination in pregnancy were enrolled to this preventive control program.A conventional strategy of screening for heterozygote was used to identify the α- and β-thalassemia traits in women and their spouses according to the standard procedures of hematological phenotype analysis which was recommended by Thalassemia International Federation (T IF).Then those suspected couples at risk were diagnosed for α- and β-thalassemia by PCR-based DNA assays.The couples at risk for severe thalassemias were counseled and offered prenatal diagnosis and termination of pregnancy in case of an affected fetus in the rights of consent and of option voluntarily.ResultsFrom January 1998 to December 2010,85 522 brides and grooms-to-be for premarital screening and 41 503 pregnant women in addition to 14 141 partners for prenatal screening were recorded,the covering rates of premarital screening and prenatal screening in the city were 92.698％ (from 1998 to 2003) and 27.667％ (from 2004 to 2010),respectively.Totally 10 726 cases were found to be the carriers of thalassemias,with 7393 for o-thalassemia (5.237％,7 393/141 166) and 3333 for β-thalassemia (2.361％,3 333/141 166).A total of 257 couples at-risk for severe thalassemias were detected including 190 for α-thalassemia and 67 for β-thalassemia.Among them,251 (97.7％,251/257) couples were performed prenatal diagnosis.During the preventive control program,a total of 72 fetuses with severe thalassemias including hemoglobin H disease were voluntarily terminated.In Zhuhai City,the average annual birth rate of fetuses with severe thalassemia was declined by 32.9％ (49/149).ConclusionsThis study has reduced effectively birth rate of perinatal infants with severe thalassemias in Zhuhai City by genetic screening and prenatal diagnosis of thalassemia in the large population of 13 years.Our summary comes out of technical proposals for prenatal screening and diagnosis,which could be take example by preventative control of thalassemia in other regions of China where are prevalent.

Objective To explore the changes in serum concentrations of irisin,vaspin and reactive oxygen species (ROS) in patients with type 2 diabetes mellitus (T2DM) and metabolic syndrome (MS),and to investigated the correlation of irisin and vaspin with clinical parameters of MS.Methods A total of 260 T2DM patients were enrolled.Age and gender were recorded,anthropometrics,biochemical parameters,and levels of irisin,vaspin and ROS in fasting serum were measured,and homeostatic model assessment of insulin resistance (HOMA-IR) calculated.Wilcoxon rank sum test,correlation analysis,Logistic regression analysis,multiple linear regression analysis and receiver operating characteristic (ROC) curve analysis were performed.Results Compared to T2DM patients without MS,T2DM patients with MS had lower serum level ofirisin [male:112.81 (86.96-191.84) μg/Lvs.156.23 (110.61-225.97) μg/L,female:141.09 (77.52-175.55) μg/L vs.172.15 (95.69-240.37) μg/L,P ＜0.01],higher levels of vaspin and ROS [male:1.13 (0.95-1.38) μg/Lvs.0.36 (0.21-0.82) μg/L,1 540 (1 250-1 860) kU/Lvs.1 020 (920-1 350) kU/L;female:1.52 (1.13-1.80) μg/Lvs.0.51 (0.47-1.08) μg/L,1 650 (1 320-1 940) kU/Lvs.1 120 (980-1 420) kU/L,P ＜0.01].In the T2DM patients,serum irisin level was negatively correlated with vaspin (r =-0.382,P ＜ 0.01) and ROS (r =-0.410,P ＜ 0.01),while vaspin was positively correlated with ROS (r =0.400,P ＜ 0.01).Multiple linear regression analyses showed that irisin was significantly correlated with body mass index (BMI),waist circumference and triglyceride,while vaspin was correlated with gender,BMI,and waist circumference (all P ＜ 0.05).Logistic regression analysis revealed that irisin,vaspin and ROS were all associated with MS (OR =0.77,95 ％ CI 0.608-0.978;OR=1.39,95％ CI 1.252-1.539;OR=1.38,95％ CI1.112-1.718,all P＜0.05).ROC analysis demonstrated that irisin and vaspin had significant area under the curve (AUC =0.931,P ＜0.01;AUC =0.777,P ＜ 0.01) for the prediction of MS.Conclusions Serum irisin level was significantly decreased,while vaspin and ROS were significantly increased in T2DM patients with MS.Irisin and vaspin were associated with clinical presentations of MS,suggesting that irisin and vaspin might be valuable predictors of MS.

Objective Through the detection of the levels of serum betatrophin,anthropometric and biochemical indices,to determine the alteration of betatrophin levels in patients with polycystic ovary syndrome (PCOS) and the relationship between betatrophin levels and metabolic indexes such as 25-hydroxy vitamin D3 [25 (OH) D3].Methods The study group was composed of 24 women with PCOS and 20 age-natched healthy women as controls.Following general physical examination of the subjects,anthropometrie measurements were performed (height,weight,waist circumference,hip circumference).Body mass index (BMI) and waist-hip ratio were calculated;25 (OH) D3,Ca2+,fasting blood glucose,fasting insulin,triglycerides,high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C) were also measured and homeostasis model assessment of insulin resistance (HOMA-IR) were evaluated in all subjects.Serum betatrophin levels were examined with an enzyme-linked immunosorbent assay (ELISA).Using independent sample T test to compare the differences between groups,the linear correlation analysis was performed to study the correlation of betatrophin with anthropometric and biochemical indices,the influencing factors of betatrophin were analyzed by multiple linear regression.Results Circulating betatrophin levels [(0.341±0.034) ng/ml vs.(0.810±0.162) ng/ml,t=4.271,P＜0.001] and HDL-C [(1.014±0.321) mmol/L vs.(1.419±0.287) mmol/L,t=2.218,P＜0.05] significantly decreased in women with PCOS compared with controls,and BMI [(27.691±4.392) kg/m2 vs.(23.310±4.781) kg/m2,t=-2.073,P＜0.05],fasting blood glucose [(5.950±0.411) mmol/L vs.(4.883±0.314) mmol/L,t=-2.142,P＜ 0.05],HOMA-IR [(4.946 ± 0.741) vs.(3.387± 0.397),t=-2.493,P＜0.05],triglycerides [(1.510± 0.848) mmol/L vs.(1.037±0.402) mmol/L,t=-2.223,P＜0.05],LDL-C [(3.431±0.479) mmol/L vs.(2.396±0.435) mmol/L,t=-2.433,P＜0.05] were obviously increased.Moreover,there was a postive correlation between betatrophin and HOMA-IR (r=0.425,P＜0.05) as well as 25(OH)D3 (r=0.577,P＜0.05) and Ca2+ (r=0.448,P＜0.05),while the betatrophin was negatively related to BMI (r=-0.451,P＜0.05),triglycerides (r=-0.454,P＜0.05),LDL-C (r=-0.551,P＜0.05).Circulating betatrophin levels were higher when 25 (OH) D3 levels were beyond 35 nmol/L in PCOS patients [(0.539±0.092) ng/ml vs.(0.199±0.031) ng/ml,t=3.072,P＜0.001],and the multiple linear regression analysis showed that the main factors affecting the levels of betatrophin were BMI (OR=-0.260),HOMA-IR (OR=0.218),25(OH)D3 (OR=0.238),and glycerol (OR=-0.162).Conclusion It is speculated that betatrophin may be a valuable predictive factor of PCOS because betatrophin is closely associated with insulin resistance,lipid metabolism disorders and the lack of 25-hydroxy vitamin D3 in PCOS patients.

Objective　To study the indication, contraindication and short-term clinical effects of stereotactic radiotherapy (X-knife) in lung cancer. Methods　A total of 56 cases (Treatment group) of lung cancer were treated with X-knife composing of the modern 3-D treatment planning system and the stereotactic body frame, and SL-75 accelerator. And another 53 cases only treated with routine external radiation served as the control. In treatment group, 42 cases received 50 Gy radiation, at 2 Gy per day, 5 d per week in the upper mediastinum, hilus of lung, below the primary focus first and then underwent X-knife for the primary focuses, and other 14 recurrent and metastatic cases received irradiation of 8-12 Gy per day, 5 d by X-knife alone. Results　The actually reexamination rate for 1, 3, 6, and 12 months after X-knife treatment in 78 focuses from 56 cases was 79.5% (62/78), 89.8% (70/78), 87.2% (68/78) and 78.2% (61/78) respectively. The focuses reappeared in 3 months after the treatment and the growth rates of focus in 3, 6, and 12 month after the treatment were 2.6% (2/78), 5.1% and 9% (7/78) respectively. There were 2 focuses received a second treatment with X-knife because no change after the first one. Both the 1-and 2-year survival rates of 36 cases of the primary lung cancer in the treatment group were higher than that of control group (89.1%, 53.9% and 77.4%, 39.6%). Conclusion　The stereotactic radiotherapy is of applicable and prospective in the treatment of lung cancer. The indications include: ①As a boost dose in case when primary focus is less than 5 cm after the first external radiotherapy may reduce the exposure of lung tissue to x-ray and the occurrence of radiation pneumonia; ②Treatment for the recurrence after radiotherapy and operation; ③Radical treatment for the metastatic focus less than 5 cm. Attention must be paid to when multiple treatment is carried on the volume of the focus, the Karnofsky scores and the general condition when stereotactic radiotherapy combined with X-knife for lung cancer.

Objective To investigate the clinical characteristics, pathogenesis, diagnosis, treatment and prognosis of acute myeloid leukemia secondary to angioimmunoblastic T-cell lymphoma. Methods The clinical data of 3 patients with acute myeloid leukemia secondary to angioimmunoblastic T-cell lymphoma including immunohistochemistry and flow cytometer analysis were analyzed retrospectively, then the literature was reviewed. Results All the 3 patients were elderly men and the initial diagnosis was angioimmunoblastic T-cell lymphoma. The 3 cases developed secondary acute myeloid leukemia in 8 months, 14 months and 34 months after treating primary neoplasms respectively. After diagnosed acute myeloid leukemia, one case died 10 months later without treatment, one case died 13 months later despite aggressive treatment and one case lost follow-up. Conclusion Angioimmunoblastic T-cell lymphoma has risk to developing acute myeloid leukemia, and there is a poor survival and the pathogenesis is unclear.

Township and Village Health Services Integration Management (TVHSIM) is an essential form of China's two-tiered health service integration plan at the township and village level.Its main purpose,also one of the target goals in China's new healthcare reform,is to gradually integrate rural health services and appropriately allocate rural health resources.This study aims to assess the village doctors' satisfaction with the TVHSIM and provide scientific base to further improve TVHSIM.A cross-sectional study was carried out in which 162 village doctors from Qinghai,Inner Mongolia and Xinjiang in western China were interviewed.Descriptive analysis,independent t-test,one-way ANOVA,Spearman rank correlation and multiple linear regression were used to analyze the difference and relevance between village doctors' personal characteristics and their satisfaction with TVHSIM and six subscales.Village doctors with different years of practice,social insurance status and essential medical knowledge level showed statistically significant differences in their satisfaction levels (all P＜0.05).Age (P＜0.05) and years of practice (P＜0.01) were negatively correlated with Drug and Medical Device Management and Financing Management.Essential medical knowledge level (P＜0.05) was negatively correlated with Operations Management as well.However,social insurance status (P＜0.05) was positively correlated with Human Resources Management and Drug and Medical Device management.Gender,age and years of practice respectively had significant influence on village doctors' satisfaction with TVHSIM (P＜0.01).In conclusion,in order to further promote TVHSIM policy in rural China,a well-rounded social insurance model for village doctors is urgently needed.In addition,the development of TVHSIM is regionally imbalanced.Efficient and effective measures aiming at rationalizing gender and age structure and enhancing essential medical training should be carefully considered.