Objective To investigate the influences of mutation at precore and basic core promoter(BCP) region in hepatitis B virus(HBV) on the immune response of specific cytotoxic T lymphocytes(CTL) in patients with chronic hepatitis B(CHB).Methods The number of specific CTL in peripheral blood mononuclear(PBMC) of CHB patients were tested by cytokine flow cytome- try(CFC) and HBV core18-27 peptide.HBV precore and BCP fragments were directly sequenced. Results Twenty-one(38.9%) samples were HBV precore G1896A mutation.Twenty-six(48.1%) samples were BCP region 1762/1764 combined mutation.Thirteen(24.1%) stains were three sites mutated simultaneously.Stimulated with HBV core 18-27 in vitro,the specific CTL level was signifi- cantly higher in the patients with G1896A mutation and BCP region mutation [(0.41?0.09)%, (0.36?0.08)%,(0.48?0.08)%,respectively]than those without mutation[(0.11?0.06)%, P

Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25％) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.

OBJECTIVE:To provide reference for the safe and rational drug use for children. METHODS:Information manage-ment system was used to investigate the use of essential medicines system variety in stock in 2015 and analyze the medication infor-mation for children in the drug instructionsin our hospital in 2015. RESULTS:Only 201 kinds of medicines belonged to children’s medicines in all the 685 kinds of medicines in our hospital. And 89 kinds (44.28%) of medicines belonged to essential medicine system among the 201 kinds of children’s medicines,in which,78 (87.60%) showed complete medication information for chil-dren;112 kinds(55.72%)of medicines belonged to non-essential medicine system,in which,38(33.93%)showed complete medi-cation information for children. The proportions of showing complete medication information for children in the essential medicines and in its chemicals,biological products,injections and oral preparations were higher than non-essential medicines,the differences were statistically significant(P<0.05). Only 41 kinds of medicines belonged to child-specific medicines among the 201 children’s medicines;62 showed complete medication information for children in the 73 kinds of essential medicines among the non-child-spe-cific medicines;only 13 showed complete medication information for children in the 87 kinds of non-essential medicines,the pro-portion of showing complete medication information for children in essential medicines among the non-child-specific medicines was higher than non-essential medicines,the difference was statistically significant(P<0.05). CONCLUSIONS:The use proportion of essential medicine system variety for children’s medicines is high in our hospital;but there are lacking of child-specific medicines and the medication information for children is insufficient. However,compared with non-essential medicines for children,the essen-tial medicines show better medication information for children in aspects of types,dosage form distribution and non-child-specific medicines,and it is suitable for children.

Objective To investigate the spatial distribution and clustering areas of Budd-Chiari syndrome in Heze City,Shandong Province,and to provide epidemiological information for further exploring the etiology and related risk factors of the disease.Methods Detailed residential addresses of 342 cases of patients (residents of Heze City) with diaphragm type Budd-Chiari syndrome diagnosed between 1995 and 2004 in Heze Municipal Hospital,Heze Shan County Central Hospital,Affiliated Hospital of Xuzhou Medical College,Shandong Provincial Hospital and Beijing Xuanwu Hospital were collected.Geographic information system (GIS) was used as a platform for data management and display.The nearest neighbor index,Ripley's K(d) function,Ripley's L(d) function and the nearest neighbor clustering method were applied to detect the spatial characters of Budd-Chiari syndrome in Heze City,Shandong Province.Crimestat 3.0 was used for spatial analysis.Results The nearest neighbor distance analysis showed that the nearest neighbor index was 0.6767 (Z =-11.4387,P ＜ 0.01).That was an aggregation at the first-order spatial scale.Within the study area,the first clustering radius of Budd-Chiari syndrome was 6.66 km,and the first clustering strength was 5.40; the average radius of the strongest clustering area was 126.61 km,and the clustering strength was 12.52,while the biggest clustering radius was larger than 222 km.After corrected by population,the gathering strength was slightly higher than that before the correction.Ten first-order hot spots were formed,and 95％ confidence interval aggregation number was 7,which meant the results were statistically significant(P ＜ 0.05),main clustering areas are in Mudan District,Shan County and Juancheng.One secondorder hot spot was gathered based on the first-order hot spot.Conclusions Spatial distribution of Budd-Chiari syndrome in Heze City,Shandong Province has showed spatial aggregation and heterogeneity.This study has a great epidemiological significance for further exploring the cause of Budd-Chiari syndrome.

Serum cortisol and plasma ACTH were determined in 10 patients with severe sepsis and 12 with septic shock on day 1,3,5 after diagnosis were made,and the data were compared with 12 control patients. To evaluate the hypothalamic-pituitary-adrenal(HPA)axis function in patients with severe sepsis and septic shock,1?g ACTH stimulation test was applied after hormone concentrations were obtained on day 1.Compared with the control patients,ACTH level was significantly higher in patients with severe sepsis and lower in septic shock(P

Objective To observe the clinical effect of digital subtraction angiography (DSA) combined with double-chamber Fogarty catheter in venous crisis after replantation of severed extremities. Methods A total of 12 cases with venous crisis after replantation of severed extremities treated by DSA combined with double-chamber Fogarty catheter from June 2011 to January 2016 were included in this study. Results Eleven cases survived after operation. Healing status of surgical incisions was stageⅠfor all patients. Of which osteofascial compartment syndrome was found in 1 case, and it was cured by surgical incision treatment; liver damage was found in another case, and acute liver damage was treated by comprehensive treatment of internal medicine. The postoperative venous thrombosis was found again in one case. The patient underwent exploratory surgery, and venous thrombosis was found again, limb partial necrosis was found, amputation was perform, and the incision was healed in Ⅰ stage. Complications including vascular rupture, air embolism, infection and sepsis were not found in all patients. Thrombosis was presented in the vein, the length of 0.6-4.2 cm. A total of 11 patients were treated with Fogarty catheters and followed up for 11 months (7 months to 29 months). The limb shape of the patient was satisfactory, blood supply of limb was improved, and capillary filling time was 1.5-3.3 s, swelling degree was improved, skin temperature was normal or lower than the limb 0.6-1.5℃, the skin color was normal, the activity improved, the feeling of recovery S0-S4 levels, the average in the S3 + level, two-point discrimination was 3-8 mm with an average of 4.5 mm. According to the evaluation criteria of the replantation function of the limb replantation of the Chinese Medical Association, 5 cases were excellent, 4 cases were good and 2 cases were poor. Conclusion The application of DSA combined with double-chamber Fogarty balloon catheter for the treatment of venous crisis shows precise localization of thrombosis, increased targeting venous branch in blood vessels, minimally invasion, quick, and satisfactory clinical results.

OBJECTIVETo study the diagnostic value and influencing factors for amplitude-integrated EEG (aEEG) in brain injury in preterm infants.

METHODSOne hundred and sixteen preterm infants with a gestational age (GA) between 27 weeks and 36(+6) weeks were enrolled as subjects. The aEEG scores of all preterm infants were obtained within 6 hours after birth. According to the diagnostic results, the 116 preterm infants were divided into two groups: brain injury (n=63) and non-brain injury (n=53). The risk factors for brain injury were evaluated using logistic regression analysis. According to the aEEG results, the 116 preterm infants were divided into two groups: normal aEEG (n=58) and abnormal aEEG (n=58). The influencing factors for aEEG results in preterm infants were determined using univariate analysis.

RESULTSThe brain injury group had a significantly higher rate of abnormal aEEG than the non-brain injury group (83% vs 11%; P<0.05). The infants in the brain injury group from two different GA subgroups (27-33(+6) weeks and 34-36(+6) weeks) had significantly lower aEEG scores than the non-brain injury group from corresponding GA subgroups (P<0.01). Logistic regression analysis showed that low GA (<32 weeks), low birth weight (<1 500 g), abnormal placenta, fetal membranes, and umbilical cord, and hypertension during pregnancy were high-risk factors for brain injury (P<0.05). There were significant differences in GA, birth weight, abnormal placenta, fetal membranes, and umbilical cord, and hypertension during pregnancy between the normal and abnormal aEEG groups (P<0.05).

CONCLUSIONSThe risk factors for brain injury are consistent with the influencing factors for aEEG results in preterm infants, suggesting that aEEG contributes to the early diagnosis of brain injury.

Birth Weight ; Brain Injuries ; diagnosis ; physiopathology ; Electroencephalography ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Logistic Models ; Pregnancy ; Risk Factors

Altitude ; Heart Rate ; Humans ; Oxygen ; Oxygen Consumption

OBJECTIVETo screen out related microRNAs in keloid tissue, and identify their effect on the proliferation of keloid fibroblasts.

METHODS8 cases of keloid tissue and 8 cases of normal skin tissue were collected as specimens. The differently expressed miRNA in keloid tissue from normal skin tissue were screened out with gene chip( Exiqon company), which was validated with quantitative real-time PCR. Then miRNA mimics was transfected into keloid fibroblasts line to stimulate high expression of mature miRNA in cells. The effect on the proliferation of fibroblasts in keloid was tested by Edu.

RESULTS(1) A total of 17 differently expressed microRNAs were found, including miR-199a-5p. (2) The expression of miR-199a-5p had been verified by qRT-PCR to be down-regulated in keloid, which was consistent with the result of array. (3) The positive rate of EdU in miR-199a-5p mimics transfected group and negative control group was (20.72 +/- 2.50)% and (27.68 +/- 4.92)%, respectively. The proliferative rate of keloid fibroblasts turned down in miR-199a-5p-transfected group (t = 2.183, P = 0.047). Besides that, the cell cycle changed after transfection. The percentage of S and G2/M phase in miR-199a-5p mimics transfected group was 33.93 +/- 1.30 and 10.87 +/- 0.80, respectively, while it was 31.39 +/- 0.79 and 9.27 +/- 0.46 in negative control group, and the difference was statistically significant.

CONCLUSIONS(1) The miRNA expression profile is different between keloid and normal skin; (2) The expression of miR-199a-5p is down-regulated in keloid and miR-199a-5p can affect the cell cycle and suppress proliferation of keloid fibroblasts. It indicateds that miR-199a-5p may be involved in regulating fibroblastic proliferation.

Cell Proliferation ; Cells, Cultured ; Down-Regulation ; Female ; Fibroblasts ; metabolism ; Gene Expression Profiling ; Humans ; Keloid ; genetics ; metabolism ; pathology ; Male ; MicroRNAs ; genetics ; metabolism

Objective: To evaluate the clinical efficacy of the Governor Vessel-unblocking and mind-regulating acupuncture method plus repetitive transcranial magnetic stimulation (rTMS) in the treatment of post-stroke insomnia. Methods: A total of 72 patients with post-stroke insomnia were randomly divided into 2 groups, with 36 cases in each group. The control group received rTMS treatment with a frequency of 1 Hz and a motion threshold value of 90％. The observation group received acupuncture with Governor Vessel-unblocking and mind-regulating method based on the rTMS treatment of the control group. The points were Baihui (GV 20), Shenting (GV 24), Yintang (GV 29), Fengfu (GV 16), Sishencong (EX-HN 1), Shenmen (HT 7), Sanyinjiao (SP 6), Shenmai (BL 62) and Zhaohai (KI 6). The treatment was performed once a day for 5 d a week followed by 2 d of rest for 4 weeks. The improvements of Pittsburgh sleep quality index (PSQI), self-rating anxiety scale (SAS) and self-rating depression scale (SDS) scores were observed after treatment, and the clinical efficacy was compared between the two groups. Results: After 4 weeks of treatment, the PSQI, SAS, and SDS scores of the two groups were all reduced, and the intra-group differences were statistically significant (all P<0.001). After treatment, the three scores in the observation group were all lower than those in the control group, and the differences between the two groups were statistically significant (P<0.05, P<0.05, P<0.001). There was a statistically significant difference between the observation group and the control group comparing the clinical efficacy (P<0.05). Conclusion: The therapeutic effect of the Governor Vessel-unblocking and mind-regulating acupuncture method plus rTMS in treating post-stroke insomnia is better than rTMS alone, and it can better improve the anxiety and depression of patients.