Objective To study the clinical features, diagnosis,and treatment of hepatolithiasis complicated with cholangiocaminoma. MethodsClinical and pathological data were analysed retrospectively on 46 cases of hepatolithiasis complicated with cholangiocarcinoma. Results The results showed that the incidence of cholangiocarcinoma in hepatolithiasis was 4%, and 33% of patients were diagnosed as cholangiocarcinoma preoperatively. Tumor occurring in left intrahepatic ducts, right intrahepatic ducts, and hilar bile duct was 72%, 9% and 20%, respectively. Only 15(33%) cases underwent radical resection with 1-, 2-, and 3 year survival rate of 100%, 67% and 25% for the 12 cases that were closely followed-up. None of 15 cases receiving palliative bile duct drainage had survived for 1 year. KG2Conclusions Patients with a history of hepatolithiasis more than 10 years are under the risk of cholangiocarcinoma.During a surgery for hepatolithiasis a thorough exploration for the possible coexistance of cholangiocarcinoma is strongly recommended.

Objective To understand the molecular mechanism underlying the epidermal growth factor receptors(EGFR)signal transduction and its feed-back regulation.Methods Two human keratinocyte cell lines,HaCaT and CHOwt,were cultured and treated with a certain concentration of different ligands,including epidermal growth factor(EGF),heparin-bounding(HB)-EGF,transforming growth factor α (TGFα)and heregulin(HER),for various durations(2,4,8,16,20 hours).After the treatment,cells were collected and protein was extracted.The amount of total and active EGFR was measured by immunoprecipitation and immunoblot assay.The internalization and down-regulation of EGFR were visualized with immunofluorescence and laser seanning confocal microscopy.Results As shown by immunoblot technique,EGF and HB-EGF continuously down-regulated the total amount of EGFRs,whereas TGFα and HER had no significant effect on the degradation of EGFRs.The activation of EGFRs was also attenuated to different extent after long-time treatment with EGF,HB-EGF and TGFα.As indirect immunofluorescenee revealed,in untreated HaCaT and CHOwt cells,EGFRs were essentially located at the plasma membrane,with a little cytosolic distribution;after ten-minute treatment with EGF,EGFRs clustered into patch-like structures which were particularly obvious in HaCaT cells,and translocated into cytoplasmic vesicles resembling endosomes (relatively apparent in CHOwt cells),while the total amount of EGFRs remained constant in these cells.The fluorescence signal from the total EGFRs decreased evidently after four-hour treatment with EGF,indicating a strong reduction in the receptors.Conclusions EGF and HB-EGF,but not TGFα or Heregulin,could down-regulate the amount of total and active EGFRs.There might be different mechanisms for the signal transduction related to EGFRs intemalization and down-regulation between HaCaT and CHOwt cells.

Objective To investigate the clinical significance of neutrophil parameters Neut‐X and Neut‐Y in the diagnosis of blood stream infection(BSI) .Methods The data in106 patients with blood stream infection and 51 cases in the healthy control group were retrospectively analyzed .The results of WBC ,neutrophil percentage ,Neut‐X ,Neut‐Y and procaclitonin (PCT ) were compared .Results Neut‐X and Neut‐Y in the BSI group were 1 393 .5 ± 33 .4 and 416 .2 ± 30 .0 respectively ,which were signifi‐cantly higher than 1 371 .9 ± 32 .7 and 391 .7 ± 23 .7 in bacterial culture negative group and 1 347 .2 ± 26 .2 and 371 .9 ± 21 .5 in the healthy control group ,the differences were statistically significant (P<0 .05) .Neut‐X and Neut‐Y had good correlation with PCT . Neut‐X and Neut‐Y levels in the Gram‐negative bacterial group were higher than those in the Gram‐positive bacterial group .Conclu‐sion Neut‐X and Neut‐Y parameters in the patients with BSI are significantly increased and can be used as the assistant diagnosis parameters of blood stream infection .

Objective To explore the effectiveness of monitoring for developmental delay (DD) and the feasibility of firsthand data reporting among 0~3 years old children. Methods A study population consisted of a cluster sample of 20907 children aged 0~3 years, from 22 communities in 5 districts in Beijing, was monitored for 1 year using questionnaires and data reports. The results were compared against the non-project communities. Results The screening rate of DD was 62.18%, higher than the non-project groups, and the rate of 0 year old group was significantly higher than the other groups. 268 children were positive screening, referral of 253 children, the referral rate was 94.4%; 97 children were diagnosed as DD with the prevalence rate of 7.46 ‰; 43.28% of the children received intervention and rehabilitation training. Conclusion The surveillance means of DD for 0~3-year-old children are effective and the firsthand data report is feasible

AIM:To detect the expression of basic fibroblast growth factor (bFGF) and matrix metalloprotei-nase 9 (MMP9) in nasopharyngeal carcinoma (NPC) and its correlation with clinicopathological features and prognosis of the patients.METHODS:The expression of bFGF and MMP9 was detected by the method of SP immunohistochemical staining in biopsy tissues of NPC patients.The relationship between the expression and the clinical significance was analyzed as well.RESULTS:In 289 cases of NPC patients, the positive rates of bFGF and MMP9 were 71.3% and 61.6%, respectively.Correlation analysis demonstrated that the expression rates of bFGF and MMP9 were both positively associated with N stage and clinical stage in NPC patients.The high expression rates of both bFGF and MMP9 were associated with poor overall survival and progression-free survival of NPC patients.Furthermore, the positive rate of bFGF was positively correlated with that of MMP9, and over-expression of both bFGF and MMP9 was correlated with the poorest survival outcomes in NPC patients.CONCLUSION:bFGF and MMP9 are over-expressed in NPC tissues and significantly associated with NPC recurrence and poor outcome.The combined interpretation of bFGF and MMP9 expression levels leads to refinement of the risks for the NPC patients and could be chosen as the prognostic biomarkers.

To study the coumarins of Anemone raddeana Regel, the compounds were separated by silica gel column chromatography and HPLC. Their structures were identified by their physicochemical property and spectral analysis. Two new compounds were isolated and identified as 4, 7-dimethoxyl-5-methyl-6-hydroxy coumarin (1) and 4, 7-dimethoxyl-5-formyl-6-hydroxycoumarin (2). The bioassays indicated that compounds 1 and 2 could significantly inhibit the proliferation of cancer cell, and showed the agonist effect on the transactivity of retinoic acid receptor-alpha (RARalpha). In addition, the two compounds had inhibitory effect against human leukocyte elastase (HLE).

ObjectiveTo observe the effects of parenteral nutrition (PN) containing ω-3 polyunsaturated fatty acids (ω-3PUFAs) on postoperative systemic inflammatory response syndrome (SIRS) in patients with obstructive jaundice. MethodsTotally 40 patients with obstructive jaundice who underwent hepatobiliary surgery in the Affiliated Hospital of Binzhou Medical College from June 2008 to October 2009 were enrolled in this study and randomly divided into the conventional PN group and PUFAs group with 20 cases in each group. The conventional PN group was provided with medium-chain/long-chain triglycerides lipid emulsion, while the PUFAs group was provided with medium-chain/long-chain triglycerides lipid emulsion supplemented with ω-3PUFAs. The two groups received PN support with equal nitrogen content and calories for 9 days. The non-protein caloric value given was 117.15 kJ/(kg · d) with 0.2 g/(kg · d) of nitrogen. Interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor-α (TNF-c) in serum were measured on the day before operation and on the postoperative day 1,3, 5, 7, and 9. Meanwhile, the incidences of SIRS and multiple organ dysfunction syndrome (MODS) were analyzed. ResultsThe levels of IL-6, CRP, and TNF-α on the postoperative day 3, 5, 7, and 9 were significantly lower in the PUFAs group than those in the conventional PN group ( all P ＜ 0.05). The duration of SIRS in the PUFAs group [(3.85 ±2.36) days] was significantly shorter than that in the conventional PN group [(5.31 ±1.47 ) days, P =0.0230]. The incidence of MODS was significantly lower in the PUFAs group ( 10％ ) than that in the conventional PN group (25％, P =0.0076). Conclusionsω-3PUFAs-supplemented PN improves the functions of liver and pancreas and alleviates acute inflammatory response in patients with obstructive jaundice.

Objective:To investigate the effect of ethyl acetate extract from Cortex periplocae (CPEAE) on apoptosis of human esophageal carcinoma cell line TE-13 and to elucidate its mechanism. Methods:Inhibitory effect of CPEAE on TE-13 proliferation was tested by MTT assay. The morphological changes of cell apoptosis were observed by Giemsa staining and transmission electron microscopy. Cell cycle and apoptotic ratio were tested by flow cytometry (FCM). The protein expression of CDK4 was observed by Western blotting.Results:CPEAE inhibited proliferation of TE-13 cells in a concentration-dependent and time-dependent manner, and its IC_(50) value was (2.443±0.005) μg/mL at 48 h (P<0.05). The characteristic morphological changes of apoptosis were observed in TE-13 cells after treatment with CPEAE under transmission microscope. A typical subdiploid peak was detected by flow cytometry. CPEAE decreased the expression of gene CDK4 in TE-13 cells. Conclusion:CPEAE can induce apoptosis of TE-13 cells. The effect is related with down-regulation of CDK4 expression.

Objective To investigate which methods can promote the serum microRNA extract production.Methods The sera of healthy persons were treated with 10％ SDS (sodium dodecyl sulfate) and/or sonication at first,then extracted miRNA by routine Trizol method,or the serum miRNA was just extracted by routine Trizol method.The contents of serum miRNA-218 and miRNA-346 were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique,and qRT-PCR relative quantities among different groups were compared to evaluate influence of different methods on microRNA extract efficacy.Results Compared to routine Trizol extraction method,the qRT-PCR relative quantities of miRNA-218 and miRNA-346 were improved when serum was treated with 10％ SDS and/or sonication at first (P ＜0.05),which was most obviously improved when treated with 10％ SDS combined with sonieation.Conclusions The method that serum was treated with SDS incubation and sonication at first can promote sensitivity of serum microRNA detection.

Objective To investigate the associations of a single-nucleotide polymorphisms (SNPs) in the interleukin (IL)-6 gene with the migraine of a Han Chinese northeastern population.Methods We collected 252 patients of the migraine attack,who were diagnosed at People' s Hospital of Liaoning Province from March 2012 to June 2013,all being Han nationality in the northeast of China.At the same time,we collected 270 healthy people over the same period who had health examination or were healthy workers of People' s Hospital of Liaoning Province.We used polymerase chain reaction and gene sequencing to analyze the genotype and allele frequencies of the-572G/C (rs1800796) IL-6 polymorphisms between migraine cases (n =252;including migraine without aura (MO) group (n =168) and migraine with aura group (n =84);the male group (n =109) and the female group (n =143)) and control group (n =270;the male group (n =116) and the female group (n =152)),as well as analyzed the genotype and allele frequencies distribution between the subgroups.Results We found higher frequencies of the distribution of the C/C genotype and the C allele of the-572G/C polymorphism in migraine cases than in control group (32.54％ (82/252) vs 19.63％ (53/270),x2 =12.061,P =0.002;55.56％ (280/504) vs 45.37％ (245/540),x2 =10.818,P =0.001),higher distribution frequencies of the C/C genotype and the C allele in female group (21.83％ (55/252) vs 12.59％ (34/270),x2 =10.187,P =0.006;33.73％ (170/504) vs 26.48％ (143/540),x2 =10.073,P =0.002).Between MO group and controls,these distribution frequencies had statistically significant difference (24.60％ (62/252) vs 19.63％ (53/270),x2 =18.172,P=0.000;39.88％ (201/504) vs 45.37％ (245/540),x2 =17.307,P=0.000).After correction by conditional Logistic regression,the frequency distribution difference of the C/C genotype between the migraine cases and controls remained statistically significant (OR =22.861,95％ CI 5.218-27.098,P =0.023).Conclusion Our data indicate that the SNPs of the IL-6 are associated with migaine and the C/C genotype increases the migraine susceptibility in a Han Chinese northeastern population.