OBEJECTIVE Gecko has been clinically used in China for many years. It has been proved that the gecko polypeptide mixture(GPM)extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci-noma(HCC)HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1)for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres-sion of apoptosis- related proteins and endoplasmic reticulum stress (ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS)generation.In this report, we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The GPM could induce ROS generation and up-regulate ERs-related proteins. CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.

OBJECTIVETo explore the effect differences between twirling-rotating reinforcing and reducing technique of acupuncture on cardiac damage in spontaneous hypertensive rats (SHR).

METHODSSixty male 11-week-old SHR were randomly divided into four groups: a model control group (group A), a twirling-rotating reinforcing technique group (group B), a twirling-rotating reducing technique group (group C) and a needle retaining group (group D), 15 rats in each one. In addition, twelve male 11-week-old Wistar rats were used as a blank control group (group E). Acupuncture was not used in group A and group E, only with grasp, capture and binding stimulation that was also adapted in the rest groups. Rats in the group B were treated with acupuncture at "Taichong" (LR 3) by twirling-rotating reinforcing technique for 1 min and then the needles were retained for 9 min; rats in the group C were treated with acupuncture at "Taichong" (LR 3) by twirling-rotating reducing technique for 1 min and then the needles were retained for 9 min; rats in the group D were treated with acupuncture at "Taichong" (LR 3) but without any technique and then needles were retained for 10 min. Before and after acupuncture, blood pressure monitor was used to measure the rats' systolic pressure and diastolic pressure every 6 days. Twenty-eight days after the treatment, HE and Masson staining were adopted to observe the status of left ventricular hypertrophy and myocardial fibrosis. ELISA method was applied to test the content of endothelin-1 (ET-1). PCR semiquantitative method was used to analyze Type I and III collagen mRNA in the left ventricular.

RESULTS(1) Blood pressure: after the treatment, the systolic pressure and diastolic pressure were both increased in the group A and the group B (P < 0.05); while the two pressures were both lowered in the group C and the group D (P < 0.05), which was more obvious in the group C (P < 0.05). (2) According to HE and Masson staining, except for the group E, the myocardial hypertrophy and fibrosis could be found in the rest groups, in which the group C was the modest, followed by the group D, while the group A and the group B were more severe. (3) Concentration of ET-1: there were differences of concentration of ET-1 among 5 groups (P < 0.05), and the concentration value from high to low was the group A, B, C, D and E. (4) Type I collagen mRNA: the difference of level of Type I collagen mRNA between group C and D was not statistically significant (P > 0.05); compared with the group A and B, the level was lower in the group C; the level was the lowest in the group E. Type III collagen mRNA: the difference between the group A and B was not statistically significant (P > 0.05); compared with the group A, B and D, the level was lower in the group C.

CONCLUSIONThe twirling-rotating reducing could reduce the systolic pressure and diastolic pressure in SHR, effectively prohibit the production of ET-1 and expression of Type I and III collagen mRNA, and it has more obvious inhibiting effect on Type III collagen mRNA. There is biological effect difference between twirling-rotating reinforcing and reducing technique.

Acupuncture Therapy ; instrumentation ; methods ; Animals ; Collagen ; genetics ; metabolism ; Endothelin-1 ; genetics ; metabolism ; Heart Ventricles ; metabolism ; pathology ; Humans ; Hypertension ; genetics ; metabolism ; pathology ; therapy ; Male ; Rats ; Rats, Inbred SHR ; Rats, Wistar

Objective To study the effects of melatonin on the expression of endothelin-1 (ET-1) and endothelial nitricoxide synthase (eNOS) in renal cortex of insulin-resistant (IR) rats. Methods Insulin-resistant rat model was established with 6-8 week ages Sprague-Dawwley (SD) rats by high-glucose diet (70% calories from glucose) for 6 weeks. IR rats models were divided into control normal group (CN, n=10), IR group (n=10) and melatonin group (MEL, n=10). Rats in MEL were affused melatonin 10mg/(kg·d) (4PM) for further 6 weeks. The protein and mRNA in rats' renal cortex of each group were assayed by immunohistochemistry and RT-PCR. Results Arterial blood pressure (ABP) and serum triglyceride (TG), free fatty acid (FFA), malondialdehyde (MDA), insulin (Ins) and homeostasis model analysis insulin resistance (HOMA-IR) in MEL were lower than that in IR (all P＜0.01). Serum high-density lipoprotein (HDL-c) and superoxide dismutese (SOD) were higher in MEL than that in IR (all P＜0.01). HE staining showed that the structure of renal cortex in MEL is approximately normal. The levels of ET-1 protein and mRNA in renal cortex of MEL rats were lower than that in IR group. The levels of eNOS protein and mRNA of renal cortex in MEL group were higher than that in IR group (all P＜0.05). Conclusion At the early stage of IR, melatonin application can ameliorate the imbalance between ET-1 and eNOS and protect the angio-endothelial function of renal cortex in high-glucose induced IR rats.

The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

OBJECTIVETo investigate the effect of different pH on rectum permeability of chlorogenic acid and geniposide.

METHODFour kinds of Reduning suppositories of different pH were separated and put into the rectum to study the suppositories in vitro and the content of chlorogenic acid and geniposide samples was determined by HPLC to calculate the permeation in 24 hours.

RESULTWith increase of pH within 2.5-7.4, the steady state flux of chlorogenic acid was increased, but the steady state flux of geniposidesamples was steady.

CONCLUSIONAdjusted the pH can increase the rectum permeability of active ingredients in Reduning auppositories.

Animals ; Chlorogenic Acid ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Hydrogen-Ion Concentration ; Iridoids ; pharmacokinetics ; Male ; Permeability ; Rats ; Rats, Sprague-Dawley ; Rectum ; metabolism ; Suppositories ; pharmacokinetics

The theoretical basis of the alcohol precipitation process control was provided, the alcohol precipitation was optimized and the relationship equation was got. The monod glycoside, loganin and paeoniflorin were used as the evaluation indexes to determine the impact factors of alcohol precipitation techniques of Liuwei Dihuang decoction by the Plackett-Burman experimental design and the levels of non-significant factors were identified. Then, Box-Behnken response surface methodology was used to research and discuss the critical process parameters influence the effect of alcohol precipitation and draw interaction between key process parameters and the correlation equation with index components. Through the establishment and solving the quadratic regression model of composite score, the optimum preparation conditions of alcohol precipitation techniques of liuwei were as follows: stirring speed was 580 r x min(-1), standing time was 17 hours, alcohol concentration was 34%, the density of Liuwei Dihuang decoction was 1.13. The response surface methodology for optimized alcohol precipitation techniques of Liuwei Dihuang decoction is. reasonable and feasible.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry

Objective To study the high-field MRI findings of gastrointestinal stromal tumors(GIST), investigate the diagnostic value of the tumor.Methods The image findings and clinical date of 45 cases with GIST proved by pathology were analyzed retrospectively.Results In 45 cases of GIST,tumors were located in the sacroanterior 2 cases, in the perineum 5 cases, in the small bowel 5 cases, in the rectum 10 cases and in the stomach 23 cases.Of 45 cases, there were 11 cases of recurrence,and 34 cases of new-onset.High-field MRI showed the tumors with inhomogeneous signal intensity, the small lesions with even homogeneous signals, the larger with non-uniform signalwhich may be cystic degeneration, necrosis, hemorrhage.After enhanced scan,the solid part of the tumor were abnormal enhancement,necrosis andcystic degeneration no enhancement.Clear or not clear of tumor boundaries andadjacent tissue pressure were prone to metastasis or intraperitoneal planting.Pathology results showed: the tumors cut surface was gray, gray-red, visible envelope.The microscopic morphology was varied, ranging from the number of spindle cells.Immunohistochemical findings were as following: CD34(+) in 43 cases;CD117(+) in 40 cases;Desmin(+) in 18 cases;Vimentin(+) in 15 cases.Conclusion High-field MRI can accurately position gastrointestinal stromal tumor, and has great value for its diagnosis and differential diagnosis.

Animal medicine is an important part of traditional Chinese medicine with a long application history in China. Systematically understanding the history of the development of animal medicine is of great significance to scientific protection and rational use of animal medicine resources. It has certain guiding significance to protection of wild resources, exploitation of new substitutes, standardization and summary of artificial breeding, and artificial reproduction technology. Taking the development of bezoar as an example, this article expounded the following four aspects:the development history of animal medicine, national animal protection, technical development, and prospect forecast by summarizing the Chinese ancient medical books and consulting the relevant laws and regulations. The entire above are about to offer new ideas for the sustainable development, the development of new medicine resources, and the development of animal medicine related preparation product.

Objective To investigate the effects of estrogen on mismatch repiar gene expression in colonic mucosa in vivo. Methods A total of 42 healthy individuals underwent colonoscopy were enrolled in the study. Half an hour before colonoscopy examination, blood sample was taken for determining the serum estradiol (E2) level. N ormal colonic mucosal tissues determined by naked eye under colonoscopy examination were taken in the right hemi colon to detect HMLH1 and hMSH2 gene expression by semi-quantitative RT-PCR and immunohistochemistry staining. Then the correlation of serum E2 levels with hMLH1 and hMSH2 expression in colonic mucosa was analyzed. Results A bimodal curve was presented for the correlation between serum E2 level in healthy individuals and hMLH1 expression in colonic mucosa. A strong positive correlation of E2 level with hMLH1 expression in normal colonic mucosa was observed when serum E2 level was more than 45 pg/ml (For mRNA, P=0. 003, r=0. 701; for immunohistochemistry positivity index, P=0. 000, r=0. 874).However there was no correlation between E2 level and hMSH2 expression. Conclusion High serum E2 level might increase the hMLH1 gene expression in colonic mucosa in vivo.

OBJECTIVE:To compare in vivo and in vitro permeation behaviors of the ethosome gels of testosterone,testoster-one propionate and testosterone undecanoate. METHODS:The ethosome gels of testosterone,testosterone propionate and testoster-one undecanoate were prepared. With cumulative permeating amount and permeation rate as the indexes,Franz diffusion cell and HPLC were employed to compare in vitro permeation behaviors of 3 kinds of ethosome gels in mouse skin. With testosterone patch as the positive control drug, electrochemistry method was adopted to detect the concentration of testosterone in plasma 0,3,6, 9,12,24,36 and 48 h after applying such 3 kinds of ethosome gels on the back of rats,and then pharmacokinetic parameters were calculated with DAS 2.0 software. RESULTS:24 h cumulative permeating amounts of the ethosome gels of testosterone,tes-tosterone propionate and testosterone undecanoate were(234.31±13.8),(175.63±41.1)and(72.60±15.3)μg/cm2,and the per-meation rates were(10.25±1.9),(7.64±1.4)and(2.96±0.8)μg/(cm2·h),respectively. The pharmacokinetic parameters of the above-mentioned three kinds of ethosome gels and the positive control drug were respectively as follows as cmax of(20.19±2.57), (17.50±2.91),(0.23±0.04),(14.97±2.12)ng/ml,t1/2Ka of(2.80±0.45),(3.36±0.59),(4.02±0.62),(4.20±0.71)h,AUC0-48 h of(13.85±1.96),(13.93±2.13),(0.35±0.07),(11.76±2.31)ng·h/ml. CONCLUSIONS:in vivo and in vitro permeation behav-iors of the ethosome gels of testosterone and testosterone propionate are fairly good.