Objective Wnt signal is very important to control the gastrointestinal mucosal proliferation, but its role in the gastric mucosal proliferation after acidified ethanol injury is not clear yet,neither is the relationship with cyclooxygenase 2 (COX-2). Methods X-gal staining was used to measure the expression of Wnt signaling; Western blot and RT-PCR were used to measure the expression of TCF4 and P-Catenin in the gastric mucosa before and after acidified ethanol injury. We also used TOP/flash plasmid to further indentify the relationship between COX-2 and Wnt signal pathway. Results After acidified ethanol injury, the expression of LacZ signal, β-Catenin and TCF4 increased only in the wild type mouse. The expression of β-Catenin and TCF4 protein increased about (3.52 ±0.52) times and (3.02 ±0.62) times separately, and the expression of β-Catenin and TCF4 mRNA increased about (19. 85 ±3.63) times and (17.82 ±4.82) times separately. Without ethanol injury, the expression of TOP/flash plasmid was inhibited by COX-2 inhibitor (NS398) about 80% and promoted by prostaglandin E2(PGE2) about 50%.After 1% ethanol injury, the expression of plasmid was inhibited by NS398 about 25% ; on the contrary,PGE2 promoted the expression about 10%. Conclusion Wnt signal in gastric mucosa is activated after acidified ethanol injury. COX-2 may work through modulating Wnt signal to control the proliferation of gastric mucosa.

There are two populations of bacteria to affect the formation of urinary stones in humanity. The first one can promote the formation of urinary stone by increasing urinary pH, decreasing concentration of urinary inhibitors , and damaging the protective urothelial glycosaminoglycan layer. The second inhibit the formation of urinary stones. These bacteria ( mainly the intestinal oxalate degrading bacteria such as Oxalobacter formigenes, lactic acid bacteria, Enterococcus faecalis etc) can decrease urinary oxalate concentration by regulating exogenous oxalate. The problems faced and the developing direction were also indicated.

Objective To evaluate the capacity of atopy patch test in diagnosis of food allergy in children with atopic dermatitis (AD).Methods Egg and milk,as the most common food allergens among Chinese children,were employed in this study.Skin prick test (SPT) and atopy patch test (APT) with fresh egg and milk were carried out in 68 children with AD.Oral food challenges in an open style were performed to confirm the diagnosis of food allergy.Resuits Of these patients,58(85.3%)were sensitive to egg,40(58.8%)to milk and 34(50.0%) to both.Of 98 patients with positive challenge,47 showed late response,10 immediate reactions.and 41 mixed reactions.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and the agreement with food challenges in diagnosis of egg/milk allergy were 96.6%/67.5%.90.0%/82.1%,98.2%/84.4%,81.8%/63.9% and 95.6%/73.5%,respectively for APT alone,37.9%/30.0%,100%/89.3%,100%/80.0%,21.7%/47.2% and 47.1%/54.4%,respectively for SPT alone.APT was found to be more sensitive in diagnosis of late-phase reactions than SPT (P<0.01).No significant difference was found in the sensitivity between APT alone and the combination of APT and SPT in parallel algorithm for diagnosis of egg or milk allergy (x2=0.509,0.549,both P>0.05) or in the specificity between APT alone and that in serial algorithm( P=1.000;x2=3.514,P>0.05).Conclusions APT is superior to SPT in diagnosis of late responses to food,and the combination of SPT and APT does not facilitate the diagnosis of food or milk allergy compared with APT alone.

Background and purpose:Hepatocellular carcinoma(HCC) is one of the most common malignancies in the world,It has a poor prognosis due to its rapid infiltrating growth and complicating liver cirrhosis.Surgical resection is considered the best curative option,the 5-year survival rate could reach 50%,especially in the patients with lesions less than 5 cm in diameter and good residual liver function.However curative surgery is feasible for only about 20% of patients.Transcatheter arterial chemoembolization(TACE) has been demonstrated that its short-term curative effect is better than the other nonsurgical treatments.However,it has not provided any survival benefit.Many studies have been done to improve the overall therapeutic effects of HCC with multi-modality approaches.The purpose of our trial was to evaluate the effects of TACE combined with 3-dimensional conformal radiation therapy(3-DCRT) for primary hepatic carcinoma.Methods:90 patients with primary hepatic carcinoma were randomly divided into two groups :45 were treated with 3-DCRT plus TACE and the others were treated with TACE alone.For TACE,80-120 mg cisplatin and 1 000 mg 5-Fluorouracil or 30 mg hydroxycamptothecine)were perfused into the hepatic arteries,then 50-100 mg adriamycin(or epirubicin) or 16 mg mitomycin C and iodized oil 10-30 ml were given to embolize the hepatic arteries,and followed by 1-2 mm gel form particles.TACE of 1-3 sessions was given to both groups.For the combined group,3-DCRT was given by 6MV X-ray.The patients with planned target volume(PTV) lesser than 512 cm~(3) received a total dose of 45-56 Gy,4.5-7 Gy per fraction,and the patients with PTV larger than 512 cm~(3) received a total dose of 45-56 Gy,2.5-4.5 Gy per fraction,5 times every week.Results:The response rates (CR+PR)of 3-DCRT plus TACE group and TACE alone group were 91.1% and 60%,respectively(?~(2)=11.79,P

Objective:To observe the effect of moxibustion on T lymphocyte subsets in peripheral blood of rats with gastric cancer. Methods:Sixty healthy Sprague-Dawley (SD) rats were adaptively fed for one week. By the random number table method, 10 rats were randomly selected as a blank group, and 12 rats were randomly selected to simulate the tumor transplantation process; after transplantation, 10 rats were randomly selected as a sham operation group. The remaining 38 rats were used to prepare gastric cancer models by gastric transplantation of the Walker-256 tumor tissue; 8 rats were randomly selected to verify the successful modeling after 7 d; the remaining 30 rats were randomly divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. From the first day of enrollment, the rats in the moxibustion group received mild moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36) (the first group) and bilateral Pishu (BL 20) and Weishu (BL 21) (the second group), and the two groups of acupoints were alternated every other day. The rats in the infrared group received infrared radiation on the stomach area and the area on the back between the T12-T13 spinous processes, the two areas were alternated every other day. Rats in the moxibustion group and the infrared group were treated for 20 min each time, once a day for 21 d. Rats in the blank group, the sham operation group, and the model group were simultaneously grasped and fixed, and no other treatment was performed. After 21 d of intervention, the rats in each group were fasted for 12 h, and blood was collected from the orbits. The numbers of CD3+, CD3+CD4+, CD3+CD8+ T lymphocytes in peripheral blood were determined by flow cytometry, and the ratio of CD3+CD4+/CD3+CD8+ was calculated. The rats were sacrificed and the thymus was dissected under sterile conditions to calculate the thymus index. Results:Compared with the blank group, the thymus index, peripheral blood CD3+, CD3+CD4+, CD3+CD8+ and CD3+CD4+/CD3+CD8+ ratio in the sham operation group did not change significantly (allP>0.05). Compared with the blank group, the thymus index of the model group was increased (P<0.05), the CD3+ and CD3+CD8+ T lymphocytes were increased (bothP<0.01), and the CD3+CD4+/CD3+CD8+ ratio was decreased (P<0.05). Compared with the model group, the thymus index of the moxibustion group was increased (P<0.01), and CD3+, CD3+CD4+ and CD3+CD8+ T lymphocytes and the ratio of CD3+CD4+/CD3+CD8+ in peripheral blood were increased (allP<0.05). Compared with the infrared group, the thymus index of the moxibustion group was significantly increased (P<0.05), the CD3+ and CD3+CD4+ T lymphocytes in the peripheral blood were significantly increased (bothP<0.01), and the CD3+CD8+ was increased (P<0.05). Conclusion:Moxibustion can significantly increase the thymus index of gastric cancer-bearing rats and activate CD3+CD4+ and CD3+CD8+ T lymphocytes in peripheral blood.

Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70 (HSP70) in gastric cancer-bearing rats. Methods: A total of 40 healthy Sprague-Dawley (SD) rats were adaptively fed for one week. The gastric cancer model was prepared by Walker-256 cancer tissue transplantation. After 7 d, 10 rats were randomly selected to verify the successful modeling, and the remaining 30 rats were divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. After enrollment, the moxibustion group received suspended moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36), (the first group of acupoints) on the 1st day, and suspended moxibustion at bilateral Pishu (BL 20) and Weishu (BL 21), (the second group of acupoints) on the 2nd day, 20 min each time, once a day. Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d. From the day of enrollment, rats in the infrared group were irradiated with the infrared radiation at the stomach area on the 1st day, and at the T12-T13 interspinous region on the 2nd day, 20 min each time, once a day, and the two locations were alternately irradiated every other day for 21 d. During the treatment, rats in the model group were intervened by grasping and fixation without treatment. At the end of the treatment, blood was collected from the inner eye orbit, and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed, the tumor volume and growth inhibition rate were measured. The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase (SP); HSP70 protein expression was determined by ELISA; HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. Results: In comparison of the model group, the volume growth of the gastric cancer in the moxibustion group was significantly restricted (P＜0.01); the volume growth inhibition rate in the moxibustion group was 37.93%; the HSP70 expression in peripheral blood and the cancer tissues was significantly increased (both P＜0.01); the expression of HSP70 mRNA and HSP70 content in gastric tumor were both obviously increased in the moxibustion group (P＜0.01); and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group. In comparison of the model group, the volume growth of the gastric cancer in the infrared group was slightly restricted (P＜0.05) with a volume growth inhibition rate of 15.89%; the HSP70 expression in the infrared group was increased significantly in peripheral blood (P＜0.01) and in the gastric cancer tissues (P＜0.05); more HSP70 was released outside of the cancer cells in the infrared group. In comparison of the infrared group, the volume growth of gastric cancer was more restricted in the moxibustion group (P＜0.05), and the HSP70 expression in the gastric cancer tissues was also higher (P＜0.05); more HSP70 was released outside of the cancer cells in the moxibustion group. Conclusion: Moxibustion and infrared treatment inhibit the gastric cancer growth in the gastric cancer-bearing rats, up-regulate the HSP70 expression in gastric cancer tissues, and promote the production and extracellular release of HSP70, and the effect of moxibustion is more obvious.

With the kernel of efficacy, "Xiaohe Silian" was a pattern and method for new drug discovery which was constituted with "metabolism-efficacy, toxicity-efficacy, quality-efficacy and structure-efficacy". Its connotation was in keeping with traditional Chinese medicine (TCM) clinical pharmacy. This paper systematically summarized the research method of new drug discovery practice process for TCM. To avoid western drug like in TCM new drug discovery, we carried out combination analysis with TCM clinical pharmacy. The correlation analysis between basic elements of "Xiaohe Silian(n) and TCM clinical pharmacy was studied to guarantee this method could integrate closely with TCM clinic from all angles. Hence, this method aimed to provide a new method for TCM new drug discovery on the basis of TCM clinical pharmacy with insisting on holistic view of multicomponent study, kinetic view of metabolic process when the curative effect occurred and molecular material view of quality control and structure-activity exposition.
Drug Discovery ; methods ; Drug Therapy ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Humans ; Medicine, Chinese Traditional

OBJECTIVETo investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL).

METHODSThe human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology.

RESULTSThe flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles.

CONCLUSIONSThe proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.

Animals ; Cell Culture Techniques ; Cell Division ; Cells, Cultured ; Collagen ; Extracellular Matrix ; Fibroblasts ; cytology ; Humans ; Rats ; Skin ; cytology ; Tissue Engineering ; methods

The argE gene from Escherichia coli coding for N-acety-L-ornithine deacetylase(NAOase), the key enzyme involved in the L-arginine biosynthesis, had been cloned in pET22b and transformed into BL21 (DE3). With 32.5% expression level in the optimal fermentation medium at 37 degrees C, most NAOase was expressed as inclusion bodies. The soluble and active proportion could be slightly increased when expressed at low temperature. The specific activity of soluble NAOase purified by Ni-NTA resin chromatography was 1193.2u/mg. The species and proportions of whole cell proteins varied with induction conditions. The inclusion bodies expressed at 37 degrees C was more pure than 22 degrees C after gradient wash with urea. Inclusion bodies could be partly refolding and reactivated by dilution and dialysis. Low protein concentration and suitable rate of oxidant/reducing agents were important to renaturation. In the optimal conditions 17.78% of Urea-denatured NAOase could be refolding and reactivated by dilution. The purified fusion protein was obtained after wash, solubilization and Ni-NTA resin affinity chromatography purification of inclusion bodies.
Amidohydrolases ; chemistry ; genetics ; metabolism ; Biocatalysis ; drug effects ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Gene Expression Regulation, Enzymologic ; Inclusion Bodies ; enzymology ; Protein Folding ; drug effects ; Recombinant Proteins ; chemistry ; isolation & purification ; metabolism ; Urea ; pharmacology

Objective:To explore the inhibitory effect of moxibustion on tumor growth and metastasis, and also its possible mechanism, in gastric tumor-bearing rats by investigating the expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF). Methods:Fifty healthy Sprague-Dawley (SD) rats (half male and half female) were routinely housed for 1 week. A total of 20 rats were randomly divided into a blank group and a sham operation group, with 10 rats in each group. The remaining 30 rats were used to make gastric cancer models by implantation of ascites-type Walker-256 cancer cells. After successful modeling, rats were randomly divided into a model group, a moxibustion group and an infrared group, with 10 rats in each group. From the day of modeling, the body weight of each group was weighed every 4 days. Warm moxibustion was alternately performed at two-group acupoints [Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36) in one group, and bilateral Pishu (BL 20) and Weishu (BL 21) in another group] in the moxibustion group. The body surface projection area of the stomach was irradiated with short-wave infrared rays in the infrared group, once a day, 20 min per time for 21 d. At the end of the treatment, the gastric tumor was completely dissected, and the tumor volume and tumor growth inhibition rate were calculated. Then the gastric tumor cell metastasis was recorded. The levels of VEGF and EGF in rat gastric tumor tissues were determined by enzyme-linked immunosorbent assay (ELISA). Results:Compared with the blank group, the body weight of the model group decreased significantly after modeling (P<0.05); compared with the model group, the rats in the moxibustion group had increased body weight during the middle and late stages (bothP<0.05). The tumor volumes of rats in the moxibustion group and the infrared group were smaller than the volume in the model group (bothP<0.05). The tumor growth inhibition rate in the moxibustion group was significantly higher than that in the infrared group (P<0.05). The case number of tumor metastasis in the moxibustion group was smaller than that in the model group and the infrared group. The VEGF level in the tumor tissues of the model group was statistically significantly higher than that in the blank group (P<0.05). Compared with the model group, the VEGF levels in the moxibustion group and the infrared group were statistically significantly lower (bothP<0.05). The EGF levels in the tumor tissues of the model group was statistically significantly lower than that in the blank group (P<0.05); compared with the model group, the EGF levels in the moxibustion group and the infrared group were statistically significantly increased (bothP<0.05). Conclusion:Moxibustion can increase the body weight, inhibit the tumor growth, invasion and metastasis in gastric tumor-bearing rats, which may be related to the regulation of VEGF and EGF expressions in tumor tissues.