AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

Objective To observe the effect of Wnt/β-catenin pathway on proliferation of human umbilical vein endothelial cell (HUVEC) using the glycogen synthase kinase 3β (GSK-3β)-targeting RNAi recombinant adenovirus vector. Methods Homologous recombination and cloning techniques were used to construct RNAi recombinant adenoviral expressive vectors specific to GSK-3β. Then, the adenovirus plasmids was transfected into HEK 293A cells to produce adenovirus and amplify the adenoviral stock. Plaque forming assay was used to titer the adenoviral stock. The GSK-3β and β-catenin protein expressions were detected by Western blot and immunohistochemistry. The proliferation of HUVEC was detected with MTr assay. Results The RNAi adenovirus vectors specific to GSK-3β were successfully produced with high titer. The expression of GSK-3β protein in HUVEC could be down-regulated efficiently by the RNAi adenovirus, along with increased β-catenin protein expression. The proliferation of HUVEC was significantly increased (P < 0.05 or P < 0.01) after infected with GSK-3β RNAi recombinant adenovirus for 3, 5, 7 days. Conclusion RNAi adenovirus is an important tool that can inhibit the expression of GSK-3β efficiently, along with increased β-catenin protein expression. Up-regulating of the Wnt/β-catenin pathway might play an important role in the proliferation of HUVEC.

Objective To explore the combination effect of unmethylated cytosine-phosphate-guanine oligodeoxynucleotide (CpG ODN) 1826 and X-rays on Lewis lung cancer in mouse and the dose response of CpG ODN.Methods The tumor-bearing mouse model was established by injecting Lewis lung cancer cells into the right infra-axillary dermis of mouse.Sixty-four C57BL /6 J mice were evenly randomized into eight groups with 8 mice each:control group,IR group,CpG OND1826 0.15 mg group,CpG OND1826 0.3 mg group,CpG OND1826 0.45 mg group,CpG OND1826 0.15 mg + IR group,CpG OND1826 0.30 mg+ IR group,and CpG OND1826 0.45 mg + IR group.On the 1st,2nd,and 9th days,CpG ODN was injected into mouse.After 3 hours of injection,the mice were start to irradiate with X-rays once a day on the 2nd-6th days,and the total dose was 12.50 Gy.Tumor growth and TGD were measured,and the apoptosis of tumor cells were examined with TUNEL.Results The Lewis lung cancer-bearing model was successfully established in all mice.Under the treatments of CpG OND1826 and irradiation,the tumor volumes were smaller than that of control group,and the tumor volumes of CpG OND1826 0.45 mg+IR group was the smallest.TUNEL results revealed that the apoptosis rate were (2.40 ± 0.51 )％ in control group,(5.62 ±0.50)％ in IR,(7.13±0.83)％ in CpG OND1826 0.15 mg,(11.63±1.06)％ in CpG OND1826 0.3 mg,(19.13 ±0.83)％ in CpG OND1826 0.45 rag,( 12.88±0.83)％ in CpG OND1826 0.15 mg+ IR,(20.57±2.37)％ in CpG OND1826 0.3 mg+ IR,and (28.17 ±3.31)％ in CpG OND1826 0.45 mg + IR group,and thus the apoptosis rate of every therapy group was higher than that in control ( t=11.15,7.91,17.82,39.48,24.73,16.61 and 17.05,P＜0.05).The apoptosis rates of CpG ODN1826 plus X-ray irradiation group were significantly higher than those in IR alone ( t =13.78,15.08 and 17.47,P＜0.05 ) or CpG ODN group (t=18.53,9.66and7.51,P＜0.05).Conclusions CpG ODN1826 can dramatically increase the efficiency of radiotherapy by inhibiting tumor growth and promoting lumor apoptosis.

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Objective To investigate the management of iatrogenic bile duct injury and evaluate the longterm efficacy.Methods The clinical data of 62 patients with iatrogenic bile duct injury who were admitted to the Peking Union Hospital from January 1982 to April 2012 were retrospectively analyzed.Of the 62 cases of iatrogenic bile duct injuries,24 were caused by laparoscopic cholecystectomy (LC) and 38 were caused by open cholecystectomy. Ten patients received non-surgical treatment, including 8 patients received percutaneous transhepatic cholangiography and drainage (PTCD) and 2 received endoscopic retrograde cholangiopancreatography (ERCP) + stent implantation. Fifty-two patients received surgical treatment,including 47 received cholecystojejunostomy,2 received cholecystoduodenostomy,3 received biliary end-to-end anastomosis. Thirty-six patients received PTCD preoperatively,and 42 received biliary stent implantation intraoperatively.Fifty patients received intraoperative peritoneal drainage to prevent postoperative peritoneal effusion or encapsulated bile collection.Results Of the 52 patients who received surgery,13 patients were complicated by more than 1 complication,including 1 case of wound infection,4 cases of cholangitis,2 cases of anastomotic leakage,2 cases of anastomotic bleeding,1 case of anastomotic occlusion, 1 case of biliary stent falling out and 3 cases of gastrointestinal diseases.Eighteen patients received cholangiography postoperatively,and 2 patients were diagnosed as with bile leakage.Fifteen patients received PTCD + biliary stent implantation,and 1 patient of them received percutaneous puncture drainage.Two patients received reoperation due to anastomotic bleeding.The mean time of peritoneal drainage for the 50 patients was (7.7 ± 2.6) days.No perioperative death was observed,and the mean operation time was (18 ± 12) days.Fifty-five patients were followed up (10 patients received non-surgical treatment,and 45 patients received surgical treatment),with a median time of 93 months.Of the 10 patients who received nonsurgical treatment,1 received reoperation due to bile leakage,3 received PTCD for the second time due to repeated cholangitis after PTCD. Of the 45 patients who received surgical treatment,6 patients had long-term complications,including 6 cases of fever,4 cases of jaundice,3 cases of choledocho-lithiasis and 1 case of bile duct canceration; 4 received reoperation at the second year after operation.Conclusions Early diagnosis of bile duct injury and operation carried out by experienced surgeons are important for a better short- and long-term prognosis.Surgical repair is the first line therapy for bile duct injury.

Chinese herbal pieces are a key factor to protecting the quality of the clinical efficacy of traditional Chinese medicine (TCM), and it is one of the basic elements of ensuring the quality of TCM and people's usage safety. However, Chinese herbal pieces has massive problem such as adulteration and counterfeit, dyeing and weighting, pesticide residues, heavy metals in excess of the standards, and all the issues are repeated excessive in the clinic treatment. These issues impacted sound development of production, management and use of TCM, but also brings common people hidden trouble for the clinical safety of medication. Protect and improve the quality of the Chinese herbal pieces demand that continue improve quality system, in-depth scientific research, and strengthen self-discipline and other factors. So it is fundamentally to ensure good quality of Chinese herbal pieces with the color, taste and shape by systematic supervision to it from the source, production, management and research, with strengthened implementation and en- forcement of the "3G".
Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; adverse effects ; Medicine, Chinese Traditional ; adverse effects ; Total Quality Management ; methods

0.05).Conclusion Febrile seizures seldomly cause severe neurological damage.

Twelve populations of Inula lineariifolia were used as materials to measure morphological characteristics, photosynthetic parameters and chemical constituents. It aims to provide a theoretical basis for germplasm resources evaluation. The results showed that I. lineariifolia had relatively rich morphological diversity, there were significant differences of morphological characteristics, photosynthetic parameters and chemical constituents among populations. There was positive correlation on morphological characteristics and P(n). Twelve populations were divided into three-type. The three populations of Xuyi, Mingguang and Fengyang were of narrower-longer leaf, bigger biomass,better photosynthetic and higher chemical constituents. Then they were classified for a similar group. It proved that the three populations were more suitable for cultivation and promotion.
Biomass ; China ; Flowers ; chemistry ; growth & development ; metabolism ; Inula ; chemistry ; classification ; growth & development ; metabolism ; Photosynthesis ; Plant Leaves ; chemistry ; growth & development ; metabolism ; Plant Stems ; chemistry ; growth & development ; metabolism

In 2009,5 523 participants of She nationality living in Ningde City of Fujian province were multistage-stratified and randomly collected,and 5 080 were enrolled in analysis eventually.The TSH screening (reference range,0.3-3.0 mIU/L) revealed biochemical hyperthyroidism in 2.4％ (2.1％ of males and 2.7％ of females) and biochemical hypothyroidism in 9.7％ (7.4％ of males and 11.4％ of females) of the participants.The prevalence of positive TPOAbs was 10.8％ (7.4％ of males and 13.5％ of females).TSH values outside the normal reference range were associated with a higher prevalence of positive TPOAbs,indicating that autoimmunity seems to be an important factor in both hyperthyroidism and hypothyroidism.

In order to explore reasonable artificial cultivation pattern of Thesium chinense, the biological characteristics and nutrients change in the process of winter dormancy of T. chinense was studied. The phenological period of T. chinense was observed by using fixed-point notation and the starch grains changes were determined dynamically by PAS-vanadium iron hematoxylin staixjing method. Soluble sugar and starch content were measured by anthrone-sulfuric acid method and amylase activity was determined by DN'S method. The results showed that the normal life cycle of T. chinense was two years. T. chinense was growing by seed in the first year, but growing by the root neck bud in the second year. During the process of dormancy, starch and soluble sugar could mutual transformation in different periods. T. chinense had sufficient carbohydrate to maintain growth and also a lot of small molecules to improve their ability to fight against adversity.
Plant Dormancy ; Plant Leaves ; chemistry ; growth & development ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; Plant Stems ; chemistry ; growth & development ; metabolism ; Santalaceae ; chemistry ; growth & development ; metabolism ; Seasons ; Starch ; analysis ; metabolism