Objective To investigate the clinicopathogenical significance of phosphorylation type of c-jun N-terminal kinase(p-JNK)and muhidmg resistance protein P-glycopretein(P-gP),multidrug resistance proteinl(MRP1)and lung resistance protein(LRP)in gastric cancer. Methods The expression of p-JNK,P-gP,MRP1 and LRP was detected in a tissue microarray containing 168 spots of gastric cancer tissue and 27 spots of normal gastric tissue by immunohistochemistry.Results The positive expression rate of p-JNK,P-gp,MRP1 and LRP in gastric cancer wag 45.8%,51.8%,45.8%and 55.4%respectively,which Was significantly higher than that in normal gastric tissue(P＜0.05).The p-JNK expression correlated with depth of invasion(P＜0.01),histology grade(P＜0.05),vessel invasion (P＜0.01),lymph node metastasis and distant metastasis(P＜0.01).The expression of p-JNK,P-gp and MRP1 in a positive relationship(P＜0.05).Kaplan-Meier analysis revealed a significant impact on survival by p-JNK,P-gp and MRPl(P＜0.05). Conclusion The p-JNK expression in gastric cancer is correlated with malignant biological behavior and may be involved in the chemotherapeutic resistance by upregulating the expression of P-g[ and MRP1.

Objective To establish an HPLC method to determine the contents of dauricine in Menispermi Rhizoma from different producing areas. Methods C18 was set as chromatographic column filler, with acetonitrile-water-triethylamine (45:55:0.1) as the mobile phase, 284 nm as the ultraviolet wavelength detection, 1 mL/min as the flow rate, 30 ℃ as the column temperature. HPLC chromatograms of eight different batches of Menispermi Rhizoma were established. Results HPLC testing conditions of Menispermi Rhizoma was established. Within 20-100 μg/mL, there was a good linear relationship between the injection volume of the reference substance and the peak area (r=0.9995). The average recovery of dauricine was 100.30%, RSD=1.000%. The contents of dauricine in Menispermi Rhizoma from different producing areas were different. Conclusion The HPLC method is with sensitivity, accuracy, precision, good reproducibility and simple operation, which can be used as detection method to determine the content of dauricine in Menispermi Rhizoma.

This study aimed at investigating the drug preparation of precise powder decoction pieces (PPDP) system,Fallopia multiflora radix preparata (FMRP) was employed in this study.Different specifications of PPDP were prepared,their extract rates were in contrast with the original pieces.Compared the quality uniformity of three batches between FMRP original slices and its PPDP extraction,the similarity of the chemical fingerprints was evaluated,and the contents of common peaks and quality uniformity were compared by relative peak areas.ITS2 sequence was taken as a DNA barcode to identify F.multiflora radix (FMR).As a result,the extract rate of PPDP was 2.5 times as much as the original slices.The average content of stilbene glucoside from the three original slices and the PPDP extraction were 3.56 ± 2.61 and 13.23 ± 0.37 mg·g-1,respectively;while the RSD were 73.28％ and 2.82％.The similarity of the fingerprints of the PPDP extraction was almost the same as that of the original slices,but the content and the uniformity of the common peaks of the PPDP extraction were significantly improved.Thus,FMR was accurately identified using ITS2 sequences.It was concluded that the PPDP considerably improve the decocting rate and quality uniformity,indicating that PPDP could save resources and improve the clinical efficacy.

Objective To explore the differences in morphology and structure of collagen fiber of the hilar bile duct between human and rat. Methods The morphology and structure of vertical and horizontal cross-section of human and rat collagen fiber of the normal and dilated hilar bile duct, and their changes under stress were observed after Masson trichrome staining. Results The morphology and structure of collagen fiber of hilar bile duet in human was similar to that in rat. The collagen fiber mainly distributed in the middle and outer layer of the hilar bile duct. The wave-like collagen fiber bundles were arranged in parallel, consistent with the longitudinal axis direction of the bile duct, and connected by the small branches. Conclusions The morphology and structure of collagen fiber of the hilar bile duct in human is similar to that in rat. The anatomical structure of the collagen fiber is adapted to its function.

Objective To investigate the expression of pancreatic thioredoxin-1 (TRX-1) in rats with acute necrotizing pancreatitis (ANP) and the effect of pretreatment of melatonin on its expression. Methods Male Spraque-Dawley rats (n = 12) were randomly divided to ANP group, melatonin group, control group with 24 rats in each group. The rats in ANP group received three intraperitoneal injections of 25 ml/kg body weight 6% L-arginine at an interval of 1 h to induce ANP. The rats in melatonin group received intraperitoneal injections of 25 ml/kg body weight 6% melatonin 30 min before ANP induction; rats in ANP group and control group received intraperitoneal injections of same amount of saline. Rats were sacrificed at 6 h, 12 h and 24 h after ANP induction. The serum level of amylase was measured and the pathological evaluation of pancreatic tissues was performed. The concentrations of malondialdehyde (MDA) and myeloperoxidase (MPO) in pancreatic tissues were measured. The expressions of TRX-1 protein were detected by immunohistochemistry and the expressions of TRX-1 mRNA in pancreatic tissues were determined by RT-PCR.Results In ANP group, serum level of amylase, MDA, MPO, TRX-1 mRNA and TRX-1 protein in pancreatic tissues were (3 012 ±1 425) U/L, (4.13 ± 1. 85)nmol/mg prot,(7.45 ± 1.26)nmol/mg prot, 0.68 ±0. 18, 66.8 ±8. 1, while they were (1 835±499)U/L, (3.03 ±2.12) nmol/mg prot, (5. 32 ± 1.06) nmol/mg prot, 0.50±0.09, 80. 29 ±8. 14, respectively in melatonin group, the values in melatonin group were significantly lower thanthose in ANP group (P < 0.05). The peak value of TRX-1 mRNA and TRX-1 protwein expressions shifted from 12 h after ANP induction in ANP group to 6 h after ANP induction in melatonin group. Conclusions The expression of pancreatic TRX-1 protein and TRX-1 mRNA in rats with ANP was significantly increased. Melatonin pretreatment could promote pancreatic tissues to express TRX-1 protein and TRX-1 mRNA, and may be protective for pancreatic tissues damages.

Objective To explore the signal transduction pathway of cyst fluid of Echinococcus granalosus in anti-parasite mechanisms through investigating the effect of cyst fluid on the expression of Toll-like receptor(TLR) in cells. Methods Changes of TLR4 and TGF-β1 expression of 7404 liver cells were detected by quantitative PCR. Results After treatment with increasing concentration cyst fluid the expres-sion of TLR4 was reduced. TGF-β1 expression of liver cells increased with the dose. TLR2 expressions in each group were very low. Conclusion Cyst fluid can increase the expression of TLR4, suggesting that the TLR4 signal transduction pathway involve anti-cyst fluid of Echinococcus granulosus. High concentrations of cyst fluid contribute to TGF-β1 expression which plays a role in immune evasion.

This study aimed at comparing the precise powder decoction pieces and market raw TCM slices of P.cacumen over the decocting quality.ITS2 sequence was adopted as a DNA barcode to identify P.cacumen.The chemical composition of the medicinal materials was characterized by HPLC fingerprints for the evaluation of the similarity of precise powder decoction pieces and market TCM slices.The concentrations of quercitrin were determined using UPLC,and the characteristic common peaks were identified.In addition,the extraction efficiency between the market TCM slices and the precise powder decoction pieces was also compared by standard decoction method.It was found that P.cacumen was accurately identified by ITS2 sequences.HPLC fingerprints showed that the extraction efficiency and similarity of the precise powder decoction pieces increased compared with the market TCM slices.However,the extraction yield rate of the precise powder decoction pieces was improved by 20％ increased in accordance with the standard decoction method,while the contents of the index component,quercitrin,presented rare increase and the decocting rates of the other chemical components little change in the study.In conclusion,it was indicated that precise powder decoction pieces improved the extraction efficiency and uniformity in comparison with TCM slices.

This study aimed at evaluating the quality of the precise powder decoction pieces (PPDP) of L.japonicae Flos (LJF) compared with the traditional commercial slices with chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,their dry extract contents were in contrast with that of commercial slices.The three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were studied by HPLC-DAD and DNA sequence alignment.As a result,the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 11.93％,which was reduced to 8.29％ after mixing and preparing into PPDP.The fingerprint showed that the slimilarity of the fringerprint of the mixed and powdered LJF was elevated with 7 common peaks.All the common peaks were increased at different levels.In conclusion,compared with traditional commercial slices of LJF,PPDP apparently improved the dissolution rate and the quality uniformity,indicating that the boiled powder of CRP obviously presented vantages in clinic.

This study aimed at evaluating the quality of precise powder decoction pieces (PPDP) of E.Folium (EF) compared with the traditional commercial slices by chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,and their dry extract contents were in contrast with that of commercial slices.The slices of EF were identified using ITS2 and psbA-trnH sequences.Three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were detected by HPLC-DAD and DNA sequence alignment.It was found that the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 15.56％,which was reduced to 6.82％ after mixing and preparing into PPDP.The fingerprints showed that the similarity of the PPDP of EF was elevated with the inceases of 10 marketed common peaks.The PPDP of EF was accurately identified by ITS2 and psbA-trnH sequences.In conclusion,compared with traditional commercial slices of EF,the PPDP apparently improved the dissolution rate and the quality uniformity,demonstrated that the boiled powder of CRP achieved obvious clinic advantages.

In this study,DNA molecular identification technology and chemical fingerprint method were adopted to evaluate the quality system of precise powder decoction pieces (PPDP) of S.suberectus dunn (SSD).ITS2 sequence was taken as DNA barcode to indentify SSD.Different specifications of PPDP were prepared,their dry extract contents were quantified in contrast with that of original slices.Three batches of SSD original slices were gleaned and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were valued by HPLC-DAD.As a result,ITS2 successfully and accurately identified the SSD in this study.The extract rate of PPDP was 15.5％,1.11 times as much as the original slices.RSD of inter-assay dissolution of cepicatechin from the original slices was 11.0％,which was reduced to 1.0％ after mixing and preparing into PPDP.The relative peak area of the 14 common peaks identified by fingerpringts were larger,while the RSD values significantly decreased.It was concluded that the PPDP of SSD improved the extraction efficiency and uniformity of the original slices,featuring quite prospective in more reasonable and scientific clinical use.