Objective To evaluate the changes in the expression of acetylated histone H3 (Ac-H3) and deacetylase silent information regulator 1 (SIRT1) in dorsal root ganglions in a rat model of negative phenotype neuropathic pain.Methods Forty-eight male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n =24 each):sham operation group (group S) and C-fiber dysfunction group (group CFD).The rats were anesthetized with 10％ chloral hydrate 3 ml/kg.C-fiber dysfunction was induced by exposing sciatic nerve to 8％ capsaicin for 30 min in group CFD.The thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were measured before and on 1,3,7 and 14 days after CFD.Six rats were then sacrificed at each time and the lumbar segments (L5) of the dorsal root ganglions were removed for detection of SIRT1 mRNA expression (by RT-PCR) and Ac-H3 and SIRT1 protein expression (by Western blot).Results Compared with group S,TWL was significantly increased at 1,3,7 and 14 days after CFD,SIRT1 mRNA and protein expression was up-regulated and Ac-H3 expression was down-regulated at 3,7 and 14 days after CFD (P ＜ 0.05),while no significant change was found in MWT at each time point in group CFD (P ＞ 0.05).Conclusion The mechanism of negative phenotype neuropathic pain is related to up-regulation of deacetylase SIRT1 expression and decreased acetylation of histone H3 in rat dorsal root ganglions.

Objective To study role of MDC/CCI22-CCR4 axis in mouse milky spots with peritoneal carcinomatosis of gastric cancer. Methods We examined the expression of CCR4 in 615 Mouse gastric cancer cell (MFC) lines by RT-PCR and Western-blot; Peritoneal metastasis model on the 615mouse was established by intraperitoneal injection of 0.2 ml MFC cells(1×104 cells). Dil fluorescence was used to observe the transfer process and section of MFC. Immunohistochemistry was conducted to detect the expression of CCR4 and CCL22 in omental milky spot; the structure of Milky spot was observed by scanning electron microscopy. Mice were randomly divided into 2 groups, namely, the saline control group (received saline) and MFC group. The concentration of CCL22 in ascitic fluid was measured in the 615 mice injected MFC after 6,8,10 days and in the saline group. Results MFC first metastasizes to the milky spot on the omentum, the expression of CCR4 and CCL22 were observered in the milky spot. The surface layer cells in milky spot consisted of discontinuous mesothelial cells and mainly macrophages and lymphocytes. The average value of CCL22 was 43 pg/ml and 364 pg/ml respectively in saline control group and MFC group.Conclusions MDC/CCL22-CCR4 axis plays an important role in the development of peritoneal carcinomatosis in mouse gastric cancer.

Objective: To investigate the laboratory,clinical and economic results of early postoperative enteral nutrition(EEN) in patients with gastrointestinal cancer.Methods: 120 patients with gastrointestinal cancer were randomly divided into three groups: EEN,early postoperative parenteral nutrition(PN), and traditional nutrition(CON)groups.The 1/3 and 2/3 target volumes were administered on the first and second postoperative days(1POD,2POD),and the full volume was given from 3POD to 8POD in EEN and PN groups.The routine formula was offerd from 1POD to 8POD in CON group.The nutritional and immune parameters were detected one day before surgery and 9POD, and the clinical and economic results were collected in three groups.Results: The levels of ALB,TF,PA,CD3,CD4,CD4/CD8 were significantly higher in EEN and PN groups than those in CON group(both P＜0.01),and the CD3,CD4,CD4/CD8 were better in EEN group than those in PN group(P＜0.05).The side effects of EEN group were similar to those of PN group(P>0.05). The recovery of bowel function was quicker in EEN group than that in PN and CON groups(both P＜0.05).The complication rate, cost of nutrition support,and cost of complication treatment were lower in EEN group than those in PN group(P＜0.05, P＜0.01, P＜0.01).Although the cost of nutrition support was lowest but the complication rate and cost of complication treatment were all highest in CON group.Conclusion: EEN is better in immune function, recovery of bowel function,complication rate, and cost-effectiveness than those in PN, and from our multiple angular viewpoints EEN is the first choice for patients with gastrointestinal cancer.

Objective To explore the role of somatosensory evoked potentials elicited by paraspinal stimulation in the detection of spinal cord lesions.Methods Patients with clinically suspected spinal cord lesions underwent somatosensory evoked potentials(SEPs)and spinal cord conduction velocity(SCCV)test using paraspinal stimulating method.96 patients aged 15 to 75 years old with suspected spinal cord lesions of various aetiologies were tested.36 patients had demyelinating disease of the spinal cord,26 had sub-acute combined degeneration of the spinal cord,19 had myelopathy,10 had acute myelitis,5 had spinal cord corhpression.Results Paraspinal stimulation elicited somatosensory evoked potentials were Performed on all 96 patients,68 of whom underwent spinal MRI as well.SEPs and SCCV were found abnormal in a high ratio in all kinds of spinal cord lesions ; in general,78 among the 96 patients had abnormal SEPs with a sensitivity of 81.25%.27 out of 36 with demyelinating disease of the spine had abnormal SEPs with a sensitivity of 75.00%,23 out of 26 with subacute combined degeneration of the spinal cord had abnormal SEPs with a sensitivity of 88.46%,8 out of 10 with acute myelitis had abnormal SEPs,15 patients with myelopathy having abnormal findings,all 5 patients with spinal cord compression had abnormal SEPs.42 of 68 patients undergoing MRI revealed to be abnormal,in which 35 patients also had abnormal SEPs.The other 26 patients had normal spinal MRI,in which 21 patients had abnormal SEPs.Conclusions Paraspinal stimulation somatosensory evoked potentials and spinal cord conduction velocity may objectively document the abnormalities of electrophysiology,which occurs earlier than those of anatomy and radiological finds, therefore it may detect the dysfunction of spinal cord at an early stage.It is even more useful in the detecting of the metabolic myelopathy,which can hardly be detected by MRI.This technique is simple,inexpensive, and maybe useful in the diagnosis for patients with suspected spinal cord lesions.

ObjectiveTo evaluate the various methods of choledochoplasty in the repair of major bile duct defects in Mirizzi syndrome.MethodsThis is a retrospective study on 3 patient with Mirizzi syndrome with a large bile duct defect.These defects were repaired by using a vascular gastric pedicle patch in our department from July 2008 to November 2011.The authors searched the domestic medical literature on surgical repair for Mirizzi syndrome in the past ten years.The patients were treated by various surgical methods,and they were analyzed according to the Csendes Classification.ResultsThere were no surgical complications in our three patients.There was one patient with a Csendes type Ⅲ,while the remaining 2 patients were with Csendes type Ⅳ.At a median follow- up of 2.5 years,no patient developed signs of chronic cholangitis.In the medical literature,there were 93 patients who were with Csendes type Ⅰ ; and 58 patients were treated by cholecystectomy only,while 35 patients were treated by partial cholecystectomy plus mucosal ablation.Of the 40 patients with type Ⅱ,29 patients were treated by direct fistula repair,9 patients by pedicle gallbladder flap and 2 patients by pedicle round ligament.Of the 20 patients with type Ⅲ,9 patients were treated by pedicle gallbladder flap.1 patient by pedicle round ligament,3 patients by pedicle gastric flap and 7 patients by Rouxen- Y hepaticojejunostomy.For the 5 patients with type Ⅳ,they were treated by Roux-en- Y hepaticojejunostomy.Of these 159 patients,postoperative complications included biliary fistula (n=1 ),upper gastrointestinal bleeding (n=1),and biliary stricture (n=1).All the remaining patients were cured.ConclusionIn patients with Mirizzi Syndrome,the choice of treatment depends on the size of the fistula.For patient with a major tissue defect in the common hepatic duct,a pedicle vascular gastric flap is a good treatment.

Aim To investigate the effect of DHA on NGF-induced neuronal differentiation of PC12 cells and explore the possible mechanism via regulating BMP pathway. Methods PC12 cells were treated with 100μg·L-1 NGF and 100 μg·L-1 NGF + 10 μmol· L-1 DHA for 3, 6 and 9 days respectively. The length and number of neurite were detected by immunofluores-cenc. DHA content was analyzed by gas chromatogra-phy in all groups. The protein expression of BMP4, BMP7 , BMPR-II and p-Smad 1/5/8 was determined by Western blot. Results The length of total primary neurite in NGF+DHA groups was obviously increased, longer than that in NGF group; DHA content in 10μmol · L-1 DHA group was higher than that in the control group;NGF+DHA groups also unregulated the protein expression of BMP4 , BMP7 , BMPR-II and p-Smad 1/5/8 . Conclusion DHA promotes NGF-in-duced neuronal differentiation in PC12 cells, which may be associated with the upregulation of BMP path-way protein.

To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral ; Animals ; Biological Availability ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Iridoids ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; administration & dosage ; blood ; pharmacokinetics

Objective To study the correlation of multi-slice CT portography (MSCTP) and digestive endoscopy in the diagnosis and evaluation of esophageal and gastric varices (EGV) caused by cirrhosis.Methods A total of 92 patients with cirrhosis were enrolled in the prospective study.All the patients were examined by endoscopy and 64-slice spiral CT scan in 4 weeks.The types,grading of EGV were observed by endoscopy and MSCTP,and Kappa conformance test was applied with the endoscopic findings as gold standard.The sensitivity,specificity,consistency,and Youden index were evaluated for the diagnosis of sophageal and gastric varices by MSCTP.Results Sixty-five patients were diagnosed to have EGV by endoscopy and 27 were negative.The positive patients included 45 patients of GOV1,19 of GOV2 and 1 patient of IGV1.MSCTP diagnosed 67 cases of EGV and 25 patients of negative results.The positive patients included 46 of GOV1,18 of GOV2 and 3 of IGV1.Two patients of IGV1 varicose veins without positive findings on endoscopy were diagnosed by using MDCTP,which revealed isolated varicose veins under the gastric mucosa.There was high consistency between MSCTP and EGV in the diagnosis of EGV (Kappa =0.732,P ＜ 0.01).The sensitivity of MSCTP was 93.8％,specificity was 77.8％,consistency was 89.1％,and Youden index 71.6％.There was high consistency between MSCTP and EGV in the classification of EGV (Kappa values were 0.743 and 0.763,P ＜ 0.01).Conclusions There is high consistency between MSCTP and digestive endoscopic in the diagnosis and classification of EGV in cirrhosis.MSCTP is superior to endoscopy in the detection of gastric varices.

Objective To investigate the relationship of TCRCα-575A/G polymorphism with anti-neutrophil antibody(ANCA) associated vasculitis in Chinese Han population. Methods 86 cases of ANCA associated vasculi-tis in Chinese Han population and 196 healthy subjects were enrolled. TCRCα-575A/G was genotyped by PCR-re-striction fragment length polymorphism (PCR-RFLP) assay. Case-control study was performed. Results No signifi-cant difference was found in either genotype distribution(AA,AG,GG) or allele frequencies between 86 patients and healthy subjects(P>0.05);But significant differences between AA group, AG group, and GG group in systolic pres-sure[(127.47±24.18)、(124.11±25.21)、(148.92±19.23) mm Hg],diastolic pressure [(75.35±14.12)、 (74.50±13.01)、(85.46±9.40) mm Hg],red blood cell count[(3.41±1.01)×109/L、(3.46±1.04)× 109/L、(2.68±0.67)×109/L] and hemoglobin [(90.45±20.69)、(100.66±29.80)、(77.61±15.81) g/L (P<0.05 for each) were found. The patients in GG group had higher blood pressure and more severe anaemia;By following the patients about (16.0±36.8) months,no statistics significance was found between groups with and without chronic renal failure in distributions and genetypes of TCRCα-575A/G (P>0.05 ). Conclusions In Chi-nese Han population,TCRCα-575A/G polymorphism might not be related to genetic susceptibility and chronic renal failure of ANCA associated vaseulitis;but G allele might be associated with more serious anaemia and hypertension.

BACKGROUND: Induced pluripotent stem cells (iPSCs) are a special type of cells with self-renewal and multi-differentiation potential, which can differentiate into intestinal organoids under certain conditions. OBJECTIVE: To explore whether iPSCs can differentiate into intestinal organoids under specific conditions in vitro.METHODS: iPSCs from B6J mice were recovered and cultured for 3 days until clone units covered about 80％ of the culture dish, and then the cells were cultured in the medium containing Activin A for 3 days until the deterministic endoderm formed. Further, the culture medium was replaced by the medium with fibroblast growth factor 4 and Wnt3A for 4 days to differentiate into the spheroids with CDX2+. After that, spheroids were collected and mixed with Matrigel,and then the mixture was dropped into the 4-well plate and cultured with Rspondin1, Noggin, epidermal growth factor, B27 and other growth factors to differentiate into intestinal organoids. Cell morphology was observed, FoxA2 and Sox17 expresson in the deterministic endoderm was detected, and CDX2, Sox9, CGA, MMP7 were measured.RESULTS AND CONCLUSION: iPSCs were cultured with Activin A for 3 days with higher cell fusion, initial differentiation and FoxA2/Sox17 expression (P < 0.05) than those of non-induced iPSCs. Spheroids began to appear at the 3rd day after culture with fibroblast growth factor 4 and WNT3A, and formed a lot at the 4th day. And CDX2 expression in spheroids was significantly increased compared with that in the deterministic endoderm (P < 0.05). Organoids gradually formed after 3 days culture, which contained all cell types of intestinal organoids, and expressions of specific markers, Sox9, CGA, MMP7, were significantly higher than those in spheroids (P < 0.05). To conclude, iPSCs can be induced to differentiate into intestinal organoids in three-dimensional niche in vitro.