Objective To discuss the clinical features and treatment methods of chronic hydrocephalus after traumatic subarachnoid hemorrhage.Methods A total of 40 cases of chronic hydrocephalus after traumatic subarachnoid hemorrhage were analyzed retrospectively.Results Eight cases improved after treatment of non-operation,the lateral ventricle external drainage was done in 32 cases of chronic hydrocephalus.Shunt tube obstruction occurred in 3 patients,with abdominal,subcutaneous and intracranial infection.The shunting operation device was pulled out and the 3 patients received operation for the second time.Two cases appeared intracranial hemorrhage,and the hemorrhage was absorbed after conservative treatment.Total Glasgow prognosis score was good in 32 cases,with mild disability in 5 cases,moderate disability in 2 cases and severe disability in 1 case.Conclusions Traumatic subarachnoid hemorrhage after chronic hydrocephalus should be early diagnosed and early treated,CT and MRI scan is the effective means to the diagnosis and differentiaion.The lateral ventricle external drainage is good method in treatment.

BACKGROUND: Two-dimensional electrophoresis (2-DE) is precise and effective for the isolation and analysis of complex protein mixtures, has become one of valuable key techniques for proteome. Mice models were as research subjects to model human disease status, one of important methods for cardiovascular disease. The establishment and optimization of 2-DE for proteomes of mice myocardium will provide basis for heart disease.OBJECTIVE: To establish and optimize a stable technique of 2-DE for proteomic analysis of mice myocardium.DESIGN: Mice were used as research subjects, observational comparative study.SETTING: Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University.PARTICIPANTS: The experiment was performed at the Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University between February and September 2004. Totally 12 male BABL/c mice of 4-5 weeks without special-pathogen free with the body mass of 12-15 g were selected, which was provided by the Experimental Animal Center of Southern Medical University.METHODS: The mice were put to death by dislocation of cervical vertebra under drugged state to obtain heart. Various protein extraction method,loading sampling buffer, strip range of IPG, sampling volume of protein,staining method and so on were compared, analyzed, scanned and digilized,and then performing image analysis. Each link in the study was established and optimized.MAIN OUTCOME MEASURES: Resolution of protein and reproducibility of the trial.RESULTS: A method that was optimal for cardiac tissue can isolate about a total of 910 protein spots on pH 4-7 gel strips. 200 μg was fit for silver stain while 1000 μg was for Coomassie stains. It was found that in the myocardium of normal mice about (920±30) protein spots were obtained in 17 em pH 3-10 L gel strip, while around (880±30) were gained in the pH 4-7 L gel strip. The mean matching rate achieves to 86. 9%.CONCLUSION: The findings of the trial showed that 2-DE technique can be effectively applied for proteomics of myocardium of mice to regulate and optimize, and relatively stable separation method of myocardial protein was established.

Objective To identify the clinical phenotypic diagnosis and gene mutation detection of two kindreds with PS deficiency. MethodsPS: A was measured by chromogenic substrate method;TPS:Ag, FPS: Ag levels were measured by ELISA method; PS gene(PROS1 gene)was detected by amplifying 15 exons and flanking intron sequences from the propositus with PCR method. PCR products were purified and directly sequenced. Results For propositus 1,PS: A was 48.6% ,TPS: Ag was 136 mg/L, FPS : Ag was 41 mg/L, PROSI gene exon 2 was in c. Heterozygous base substitutions was detected in C121T locus, which led to Arg-1Cys (R-1C) heterozygous roissense mutation encoded in PS proteins. For propositus 2, PS: A was 29.2%, TPS: Ag was 83 mg/L, FPS: Ag was 26 mg/L, PROSI gene exon 14 was in c. Heterozygous base substitutions was identified in CI687T locus, in which Gln.522Stop heterozygous nonsense mutation was encoded in PS proteins. Conclusions c. C121T is a novel mutation locus detected in PROS1 gene. This heterozygous mutation could lead to type Ⅱ PS hereditary deficiency, while c. C1687T heterozygous mutation could bring about type Ⅰ PS hereditary deficiency.

Objective To construct the pseudovirus containing nucleic acid(NA)fragments of Marburg virus,Zaire Ebola virus,Sudan Ebola virus,Lassa fever virus and Yellow fever virus by using a lentiviral vector system in order to provide a reference standard for the detection of the five viruses.Methods The gene fragments of the above five viruses were synthesized in vitro,connected into a single gene by fusion PCR technique,and cloned into lentiviral vectors with its auxiliary vector.After co-transfecting into 293T cells,the supernatants were collected on 48 h and 72 h post transfection. The naked NA was cleaned from the supernatants using DNase and RNase digestion before pseudotype virus was purified and concentrated.After the NA of the pseudotype virus,were extracted normal PCR and real-time PCR were conducted. Results Sequence analysis showed that the five target genes in vitro synthesis were properly connected and inserted into lentivirus vectors.Using the NA of the pseudotype virus as the template,both normal PCR and real-time PCR could sensitively amplify the target gene with the primers and probes of the above five,viruses respectively.The result indicated that the pseudovirus particles containing the five kinds of hemorrhagic fever virus target genes were successfully packaged. Conclusion The pseudovirus particles containing gene fragments of five viruses are constructed,which can be used as a common reference standard for NA detection.

Objective To observe the effects of human EP cells (hEPCs)transplantation on the function recovery of rats with spinal cord injury.Methods 24 SD rats (5 ones died in the process of operation)were firstly completed transection of T10 segment of spinal cord by using quick knife slices to complete animal model.19 rats after spinal cord transection were randomly divided into:the experimental group (10 cases)and the control group (9 cases). The experimental group with spinal cord injury was injected 7.5μL suspension of hEPCs (human Endothelial Progeni-tor Cells).Adjacent area of spinal cord injury in control group was injected equal amount DMEM(Dulbecco minimum essential medium.Postoperative intraperitoneal injection of cyclosporine A,an immunosuppressant,was administrated daily.At postoperative 1,2,4,6,8 weeks Basso Beatlie Bresnahan (BBB)score were administrated to record the neural function recovery of lower limbs in SCI rats.The transection spinal cord were removed from rats 8 weeks after operation to detect transplanted survival hEPCs in transection spinal cord of ratsby immunohistochemical and hybridization technique. Results 2 weeks after SCI model established,the BBB scores in the experimental group and the control group showed no statistically significant difference(t =0.61,0.69,all P >0.05).4 weeks postoperatively,the BBB score of the experimental group (1.67 -0.71)points was more than that in the control group (1.11 -0.60),the difference was statistically significant(t =3.16,P <0.05).Untill 8 weeks scores were respectively the (4.00 -1.41)of the experimental group and the (1.44 -0.89)of the control group,the difference was statistically significant(t =5.384,P <0.05). Finally it was concluded that 4,6,8 weeks after SCI,hind limbs function of the experimental group was significantly better than that of the control group,the difference had statistical significance (P <0.05).Hybridization and immuno-histochemical detection showed that the transplanted hEPCs not only live in the host but in partial hEPCs chimeric to SCI in rats of spinal cord and its vascular system.Conclusion After transplantation,hEPCs can survive,differentiate into vascular endothelial cell,and improvement spinal cord function recovery as compared with control group.

Objective To establish a one-step recombinase polymerase amplification (RPA) method for pathogen screening and rapid detection in the field targeting for five hemorrhagic fever related viruses (Zaire ebola virus, Sudan ebola virus, Marburg virus, Lassa virus and Yellow fever virus). Methods The specific nucleic acid (NA) fragments of each virus were selected as target genes by genome sequence analysis, and the primers and probes for RPA assays were designed according to the sequence. A series of diluted template genes were used for RPA detection to determine the sensitivity. The hemorrhagic fever-related viral nucleic acids were used for RPA detection to determine the specificity. The amplification experiments were carried out at different temperature ranging from 37℃ to 42℃ to validate the reaction temperature range. Results The RPA reaction systems of the five hemorrhagic fever viruses could effectively amplify the target genes, the sensitivities were between 1.5×102 and 1.5×103 copies. No cross reactions existed with the other hemorrhagic fever-related viral genes. Meanwhile, RPA assay could effectively amplify the target genes at 37-42℃. Conclusion The isothermal RPA assays of five hemorrhagic fever viruses are established, which may amply target genes fast and react at a wide temperature range, and be potentially useful for in field pathogens detection.

Background Human LECs can express telomerase activity,which participates in the formation of cataract.It is reported that estrogen can increase the expression of telomerase activity in human endometrial cancer and breast cancer cells and play an important role in promoting proliferation and anti-apoptosis,but whether estrogen exerts its role on human LECs is still unclear.Objective This study aimed to investigate whether β-estradiol (β-E2) can increase the telomerase activity of human LECs and the influence of β-E2 on proliferation and apoptosis of human LECs.Method Human LECs line was cultured and passaged in vitro,and 1×10-6 mol/L β-E2 was added in the medium for 0,6,12,24 and 48 hours,and reverse transcription PCR was used to determine the optimal time of the expression of human telomerase reverse transcriptase (hTERT) mRNA in the cells.Cultured cells were divided into five groups.The cells in the blank contol group were cultured in the routin medium.Ethanol of 0.1％ was added in the solvent control group,and 1 × 10-8,1 × 10-7 or 1 × 10-6 mol/L β-E2 was added in the medium in different contents of β-E2 groups,respectively.The relative expression level of hTERT mRNA in different groups was detected by reverse transcription PCR.Telomere repeat amplification protocol (TRAP)-ELISA was employed to determine the telomerase activity.The proliferative value of the cells was assayed by cell counting kit-8,and the apoptosis rate of the cells was examined by Hoechst33258 staining.Results The optimal time of β-E2 to rise the expression of hTERT mRNA (absorbance) was at 24 hours under the 1×10-6 mol/L.The relative expression levels of hTERT mRNA in the cells were 0.477±0.015,0.712±0.013 and 0.914±0.031 in the 1 ×10-8,1 ×10-7 and 1 ×10-6 mol/L β-E2 group,which were signifincatly higher than 0.428±0.010 in the blank control group and 0.426±0.010 in the solvent control group (all at P＜0.05).The telomerase activity values (absorbance) were 0.711 ±0.015,0.941±0.010 and 1.249±0.047 in the 1×10-8,1×10-7and 1×10-6 mol/L β-E2 group,which were higher than 0.535±0.013 in the blank control group and 0.543 ±0.013 in the solvent control group (all at P =0.000).The proliferantive values of the cells (absorbance) were significantly raised in the 1 × 10-8 mol/L β-E2 group compared with l × 10-7 and 1 × 10-6 mol/L β-E2 group (both at P =0.000),and no significnant difference was found in the proliferetive values between the blank control group and the solvent control group (P =0.718,0.856).The apoptosis rates of the cells in the 1 × 10-8,1 × 10-7 and 1 × 10-6 mol/L β-E2 group were lower than those in the the blank control group and the solvent control group (all at P=0.000),and there was no significant difference between the blank control group and the solvent control group (P =0.777).No obvious correlation was found between the HLECs preliferative values and hTERT mRNA expression levels or telomerase activity values (r=-0.299,P=0.278;r=-0.157,P=0.576).However,significantly negative correlations were seen between apoptosis rates and hTERT mRNA expression levels or telomerase activity values (r =-0.975,P=0.000;r=-0.981,P=0.000).Conclusions β-E2can increase the activity of telomerase in human LECs,and high dose of β-E2can inhibit apoptosis,but it dose not promote proliferation.

Aim To determine the function of nuclear factor-κB ( NF-κB ) in immunological liver injury of rat model and its effect on CYP2E1 expression, content and metabolic activity. Methods The immunological liver injury rat model was prepared by injection of Ba-cillus Calmette Guérin ( BCG,125 mg · kg-1 ) for 14 days. The hepatic tissue injury was revealed by hema-toxylin and eosin ( HE ) method and serum concentra-tion of alanine aminotransferase( ALT) , aspartate ami-notransferase ( AST ) respectively. CYP450 total con-tent in hepatic homogenate was determined by spectro-photography. The expression of CYP2E1 protein was detected by Western blot analysis. The enzyme kinetics of CYP2 E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography ( HPLC ) as-say. Results The results showed that BCG-pretreat-ment ( 125 mg · kg-1 ) significantly increased the weight of liver and spleen, serum levels of ALT and AST(P<0. 01) , and decreased CYP2E1 expression, content and metabolic activity ( P <0. 05 ) . Adminis-tration of ammonium pyrrolidine dithiocarbamate (PDTC) (50, 100, 200 mg·kg-1) reversed the a-bove hepatic injury stimulated by BCG in vivo. Moreo-ver, PDTC dose-dependently inhibited the down regu-lation of CYP2 E1 ( P<0. 05 ) . Conclusion Passiva-tion of NF-κB can inhibit the down regulation of CYP2 E1 in liver tissue of immunological liver injury rats;NF-κB may be involved in CYP2 E1 down-regula-tion.

Objective To investigate the correlation between 28-day prognosis and red cell distribution width (RDW) in sepsis patients.Methods This was a prospective observational study.Two hundred and thirteen sepsis patients were consecutively selected,and the patients were divided into 2 groups according to RDW:normal RDW group (RDW ＜ 0.15,160 cases) and high RDW group (RDW≥0.15,53 cases).The general conditions,acute physiology and chronic health evaluation (APACHE) 1Ⅱ score,sequential organ failure assessment (SOFA),hypersensitive C reactive protein (hs-CRP),procalcitonin (PCT),arterial blood lactic acid,liver function injury,renal function injury and 28-day mortality were compared between 2 groups.The independent risk factors of 28-day prognosis were analyzed by multifactor Logistic regression analysis.Kaplan-Meier survival analysis was used to draw the 28-day survival curve,and the survival rate was compared between 2 groups by log-rank test.Results The 28-day mortality in high RDW group (35.8％,19/53) was significantly higher than that in normal RDW group (17.5％,28/160),and there was statistical difference (P =0.007).RDW ≥0.15 was the independent risk factor of 28-day death in sepsis patients (OR =2.634,95％ CI 1.316-5.273,P =0.006).After adjusted by gender,age and other relative factors,RDW≥0.15 was the independent risk factor of 28-day death in sepsis patients (OR =2.895,95％ CI 1.155-7.252,P =0.023).The 28-day accumulative survival rate in high RDW group was significantly lower than that in normal RDW group (50.5％ vs.63.0％),and there was statistical difference (P =0.014).Conclusion The high RDW in sepsis patients is the independent risk factor of 28-day death,and RDW ≥0.15 shows an important predictive value in the prognosis of sepsis patients.

Objective To analyze the relationship between the abdominal fat and the progress of carotid atherosclerosis in the subject ageing 51-100 years.Methods 140 subjects receiving health examination in the department of health of Peking Union Medical College Hospital from 2015 to 2016 were included in the research.The abdominal fat area and distribution were calculated according to abdominal CT,and the progress of atherosclerosis in carotid artery was determined by ultrasound.Results In the population of 51-100 years old,there were no statistically significant difference in abdominal fat area and distribution among carotid artery plaque thickening group,arteriosclerosis nori-progressing group and plaque reducing group;In the population of 51-80 years old,the total abdominal fat was significantly higher in carotid artery plaque thickening group than in arteriosclerosis non-progressing group and plaque reducing group (P=0.05,P =0.03),abdominal visceral fat area also increased,but no significant difference was found (P＞0.05),and no significant differences in abdominal fat distribution was found (P＞0.05.Conclusion The less total area of abdominal fat is,the slower the progress of atherosclerosis in carotid artery is in the population of 51-80 years old.