Objective To analyse the etiology and clinical characteristics of 26 critically ill children with severe hand foot and mouth disease (HFMD) of Shanxi province in 2010.Methods We retrospectively analysed the clinical data of 26 children with severe HFMD from Mar to Sep 2010.Nucleic acid of enterovirus 71 ( EV71 ) and Coxsackie virus A 16 ( CoxA16) were detected in 20 out of 26 children with HFMD by reversed real time polymerase chain reaction (rRT-PCR),and the whole VP1 gene of EV71 deriving from 6 different areas of Shanxi province was amplified,sequenced,and compared with strains from other areas in china.Results EV71 nucleic acid were positive in 18 out of 20 children,while the other two were negative for EV71 and CoxAl6.Among all the critical cases,20 cases (76.9％) occurred in Weinan area,four in Xianyang area,and two in Xi'an urban area.Compared with those of Fuyang Anhui,Hong Kong China,Guangzhou,Shenzhen,Shandong,Beijing,the homology of the whole VP1 gene sequence from 6 strains of Shanxi area was 96％ ～ 100％.Most of the critical children were under 3-year-old,and the incidence rate of male children was higher than that of female children.All affected children had persisted fever,poor energy,hyperarousal,hypersomnia,and limb shaking.Meanwhile their peripheral blood leukocytes,C-reactive protein and blood glucose were markedly increased,but renal injuries were rare.Eighteen children clinically recovered on discharge,among which 2 cases had sequelae of limb activity obstacle,and 8 cases died.Conclusion Weinan is the area with the highest incidence rate of critical HFMD cases in Shanxi Province,and the major etiological organism is EV71,which is highly homologous to EV71 found in other regions of mainland China.As many cases are in dangerous condition,thus early identification and intervention could inhibit the disease progression,and play a key role in reducing the mortality.

A novel high performance liquid chromatographic method was developed for the determination of 4-O-methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation.Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins.A reverse phase column of SHIMPACK VP-ODS (150 mm× 4.6 mm,5.0 μm) was used to separate 4-O-methylhonokiol in the plasma samples.The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0.012- 1.536 mg/L.The good extraction recoveries were obtained for the spiked samples (84.7％,89.3％ and 87.7％ for low,middle and high concentrations of added standards,respectively).The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6％ to 13.5％.The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-O-methylhonokiol showed a two-apartment open model.This work developed a sensitive,stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.

Objective: To explore the effect of resveratrol (Res) on angiogenesis of human umbilical vein endothelial cells (HUVEC) with the possible mechanisms in vitro.
Methods: The HUVECs were cultured in 6 groups.①Control group, HUVEC were cultured with high glucose in DMEM,②Res group, the cell were cultured with Res at different concentrations,③Res with PI3K blocker LY294002 (Res+Blocker 1) group, ④Blocker 1 group, HUVEC were cultured with LY294002 alone, ⑤Res with eNOS blocker L-NAME (Res +Blocker 2) group and ⑥Blocker 2 group. The effect of Res on HUVEC proliferation was detected by CCK-8 kit, the protein expressions of p-Akt, p-eNOS were examined by Westin blot analysis, nitric oxide (NO) level was measured by nitrate reduction method and the endothelial cell migration was assayed by transwell chamber method.
Results: ① Compared with Control group, HUVEC proliferation increased in Res (1, 5μmol/L ) group, P<0.01, the proliferation in Res (5μmol/L) group was higher than those in Res (0.2, 10, 20μmol/L) group, P<0.01, while Res (20 μmol/L) group could inhibit the proliferation P<0.01. ②Compared with Control group, Res (5μmol/L) group had the higher protein expressions of p-Akt, p-eNOS, P<0.05-0.01, higher NO level, P<0.05.③Compared with Res group, Res+Blocker 1 group had lower expressions of p-Akt, p-eNOS, P<0.01, lower NO, P<0.05; the expressions of p-Akt, p-eNOS and NO level were similar between Res+Blocker 1 group and Blocker 1 group, all P>0.05.④Compared with Control group, the cell migration and tubing formation were higher in Res (5μmol/L) group, P<0.01;compared with Res group, the cell migration and tubing formation were lower in Res+Block2 group, P<0.01.
Conclusion: Res could up-regulate NO level via activating PI3-K/Akt/eNOS signaling and therefore, improving the proliferation, migration and tubing formation of HUVEC in vitro.

Objective:To compare the osteogenic differentiation of dental pulp stem cells(DPSCs)induced by osteopontin(OPN)and mineralizing culture medium(MCM).Methods:DPSCs were cultured with OPN(OPN group)and MCM(MCM group)respectively. The morphology of the DPSCs were observed under inverted microscope.The mineralize nodules were observed by alizarin red staining. RT-RCR was used to detect the mRNA expression of bone sialoprotein (BSP),Runt-related transcription factor 2(Runx-2),osteocal-cin(OCN)and collagen-1(Col-1).Results:Similar number of mineralized nodules was found in the 2 groups(P >0.05)after 28 day culture.The mRNA expression level of BSP gene in OPN group was higher than that in MCMgroup(0.864 ±0.112 and 0.514 ±0.068, P <0.05),while the expression level of Runx-2 gene in OPN group is lower than that in MCMgroup(0.186 ±0.017 and 0.324 ±0. 058,P <0.05).The expression level of Col-1 and OCN genes in both groups were similar(P >0.05).Conclusion:The capabilities of OPN and MCMin inducing osteogenic differentiation of DPSCs are similar.

Objective:To investigate the expression of tissue factor pathway inhibitor-2 (TFPI-2) and synuclein gamma (SNCG) in esophageal cancer and their correlation with local invasion, lymph node metastasis and apoptosis. Methods:The expression of TFPI-2, SNCG, and matrix metalloproteinase-9 (MMP-9) was detected by immunohistochemical SP methods in 82 cases of esophageal cancer tissues, 20 cases of atypical hyperplasia tissues, and 54 cases of para-cancerous tissues. The apoptosis of esophageal cancer cells was detected by TUNEL staining and apoptosis index (AI) was calculated. Results:The positive rates were 30.4%, 60.0%, and 87.0% for TFPI-2 protein and 63.4%, 30.0%, and 3.7% for SNCG protein in the tumor tissues, atypical hyperplasia tissues,and tumor-adjacent normal tissues, respectively. There was a significant difference between the three groups(P<0.01). The positive expression of TFPI-2 and SNCG correlated with the lymph node metastasis, invasion depth, TNM staging, and differentiation degree of esophageal cancer (P<0.01), but did not correlate with age at surgery, gender, tumor location, and pathologic classification(P>0.05). The expression of TFPI-2 and MMP-9 was negatively correlated (r=-0.636, P=0.000). The expression of SNCG and MMP-9 was positively correlated(r=0.393,P=0.000). AI was related with TFPI-2 and SNCG expression (P<0.05). Conclusion:TFPI-2 not only inhibited the expression of MMP-9 but also induces apoptosis of esophageal cancer to prevent tumor invasion and metastasis, however, SNCG plays a contradictory role in cancer development. TFPI-2 and SNCG might serve as new tumor markers and the new targets for tumor gene therapy.

A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins. A reverse phase column of SHIM- PACK VP-ODS （150 mm ×4.6 mm, 5.0 μm） was used to separate 4-O-methylhonokiol in the plasma samples. The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0. 012 - 1. 536 mg/L. The good extraction recoveries were obtained for the spiked samples （84.7%, 89.3% and 87.7% for low, middle and high concentrations of added standards, respectively）. The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6% to 13.5%. The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-0-methylhonokiol showed a two-apartment open model. This work developed a sensitive, stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.

Objective To explore the clinical value and safety of early enteral nutrition support in patients after liver transplantation.Methods We retrospectively analyzed the clinical data of 86 cases who used early enteral nutrition support therapy after liver transplantation between January 2008and October 2011.All of patients were uproot the gastric tube at the first day after the operation,and gradual to the normal diet.The patients who used parenteral nutrition support therapy were as the control group(n＝112).Then we compared the data of patients in the two groups.Results The early enteral nutrition is more useful to the patients after liver transplantation than intravenous nutrition [In the seventh day after the operation,the control group's ALT was (45.2 ± 12.9) U/L,AST was (40.2±9.4) U/L,ALBwas (35.6±2.5) g/L,P＜0.05].The early enteral nutrition also can decrease hospital stay and hospital costs [(14.2±3.4) d,P＜0.05].Conclusion The early enteral nutrition is useful and safe to the patients after liver transplantation.

Objective To investigate the value of partial hepatectomy and vascular resection in the treatment of hilar cholangiocarcinoma. Methods Seventy four patients with hilar cholangiocarcinoma who underwent hepatectomy of Chinese People' s Liberation Army from January 2008 through December 2011 were analyzed retrospectively.Results Of the 74 patients,33 underwent radical resection and 19 palliative resection,22 received internal or external drainage.In the radical resection group,the median survival time was 27 months,and the overall survival rate at 1,2 and 3 years were 79％,64％ and 49％.In the palliative resection group,the median survival time was 14 months and the overall survival rate at 1,2 and 3 years were 56％,25％,and 19％.In the drainage group,the median survival time was 9 months and the overall survival rate at 1,2 and 3 years were 23％,15％,0.Conclusions Hepatectomy combined with hilar vascular resection helps increase survival rate of patients in radical excision of hilar cholangiocarcinoma and Surgical resection is the most elective method for treatment of hepatic hilar cholangiocarcinoma,and the radical resection might improve the prognosis of the patients with hilar cholangiocarcinoma.

Objective To evaluate the feasibility,safety and efficacy of laparoscopic radiofrequency ablation (RFA) therapy for hepatocellular carcinoma (HCC).Methods Sixty-eight cases of liver cancer lesions were underwent laparoscopic radiofrequency ablation,and their postoperative recovery state,focal necrosis rate were observed.Results All the 68 cases were successfully performed operation,114 lesions were treated including 20 missed lesions at preoperative imaging diagnosis.There were no serious postoperative complications,the average hospital stay was (2.5 ± 1.2) days,focal necrosis rate 3 months after operation was 85.9％,lesion recurrence rate 6 months after operation was 12.2％,the 1-year survival rate was 76.47％.Conclusions Laparoscopic radiofrequency ablation in treatment of hepatocellular carcinoma has high security,few complications,short hospital stay and remarkable clinical effects.It's well worth clinical outreach.

BACKGROUND:The induced concentration for osteoblasts is often introduced as reference to induce odontoblast differentiation. However, there are no reports on other concentrations. OBJECTIVE: To observe the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein during low-dose β-glycerophosphate-induced differentiation of dental pulp stem cels into odontoblasts. METHODS:Human dental pulp stem cels were isolated and cultured, and then induced by different concentrations of inducing solution to differentiate into adipocytes and osteoblasts, which could verify the multi-directional differentiation ability of human dental pulp stem cels. Under 5 mmol/L β-glycerophosphate, dental pulp stem cels differentiated into odontoblasts. At 7, 14, 21, 28 days of culture, RNA samples were extracted from dental pulp stem cels in each group, and reverse-transcription PCR was used to detect the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein. Mineralized nodules were detected by alizarin red S staining to show the successfuly osteogenesis induction. RESULTS AND CONCLUSION: Human dental pulp stem cels could be induced to adipocytes and osteoblasts. The results of reverse-transcription PCR showed that the dental pulp stem cels under 5 mmol/L β-glycerophosphate could increase the expression of dentin matrix protein-1 and dentin sialoprotein, but downregulate the expression of matrix extracelular phosphoglycoprotein at 7, 14, 21 days. At 28 days of culture, dental pulp stem cels were al successfuly mineralized detected by alizarin red S. There were some red mineralized nodules. These findings indicate that the 5 mmol/L β-glycerophosphate can induce the differentiation of dental pulp stem cels into odontoblasts successfuly, up-regulate the mRNA expression of dentin sialoprotein and dentin matrix protein-1, and meanwhile down-regulate the mRNA expression of matrix extracelular phosphoglycoprotein.