Objective To investigate the relationship between HBV genotypes and YMDD motif mutations or BCP mutations in Xiangtan of Hunan Province. Methods HBV genotypes, YMDD motif mutations and BCP mutations were analyzed in 952 HBV infected patients. Results HBV genotyping showed that 698 HBV type B patients and 115 HBV type C patients accounted for 73.32% and 12.08% respectively of all the participants. The rest 139( 14.60% )were genotype B and C mixed infection( B + C ). The analysis of YMDD motif mutations showed that 844 YMDD wild-type which accounted for 88.66% of all the subjects and the remainder were YMDD mutation types, of which 54( 5.67% ) carried YVDD, 53( 5.57% ) YIDD,and 1 YVDD and YIDD mixed infection. Basic Core Promoter mutations showed that 1762A/1764G ( wild type )accounted for 70.59% and 1762T/1764A( mutant ) accounted for 19.75%. The rest 92 patients were 1762T/1764A and 1762A/1764G mixed infection. This study showed no significant difference in the rate of YMDD mutation( 10.04% vs 10.43% ,χ2 =0.017,P＞0.05 ) ,but a significant difference in the types of YMDD mutation(χ2 = 4.836, P ＜ 0.05 )between HBV types B and C. The YVDD mutation was more commonly seen in genotype C( 9.57% ) than in genotype B( 5.88% ). The BCP mutation rate showed a significant difference( 27.36% vs 46.09%, χ2 = 16.478, P ＜ 0.01 ). Genotype C was more frequent than genotype B. The BCP mutation rate showed no significant difference between YMDD Wild-type and YMDD mutation types( 28.67% vs 35.51%, χ2 = 2.139, P ＞ 0.05 ), but most of BCP mutations happened in YVDD mutant type( 61.11% ). Conclusions ( 1 ) The predominant HBV genotypes in Xiangtan were genotype B and genotype C, the major genotype was type B, which display the characteristics of epidemiology in Southern China. ( 2 ) Determination of HBV genotypes before lamivudine therapy was probably not an important pretreatment investigation to predict antiviral responses. ( 3 ) Detection of HBV genotypes, YMDD motif mutations and BCP mutations will contribute to the correct evaluation of prognosis and timely proper management of HBV patients.

Objective: To further study the effects of As 2 O 3 in various concentrations on the esophageal carcinoma cell line. Methods: The esophageal malignant transformed epithelial cells (SHEEC1) were induced by HPV18 in synergy with TPA in our laboratory. The cells cultured in flask and 24 wells plate were treated by As 2 O 3 with concentrations of 1, 3 and 5 ?mol/L for 2, 4, 8, 16, 24 h respectively. The morphologic changes of cells were observed under election microscopy. The mitotic index (MI) of living cells was calculated by phase contrast microscopy and the cells with TdR uptake were examined by autoradiography. The proliferative index (PI) and apoptotic index (AI) were assayed by flow cytometry. Results: A low dosage of As 2 O 3 (1.0 ?mol/L) enhancing the protiferative rate of SHEEC1 was demonstrated with TdR uptake, MI and PI increased. The high AI and low PI were found in the high concentrations (3 and 5 ?mol/L)of As 2 O 3 . The morphological changes of apoptosis and necrosis were found in 24 h after As 2 O 3 in high concentrations (3 and 5 ?mol/L) administrated. Conclusion: The effects of As 2 O 3 in various concentrations are different. Low concentration of As 2 O 3 promotes the proliferation of the esophageal carcinoma cells by increment of DNA synthesis. In high concentration of As 2 O 3 apoptosis and necrosis are induced.

Objective To investigate effect of mild hypothermia on changes of somatosensory evoked potential (SEP) and synaptophysin mRNA level after traumatic brain injury (TBI) and determine hypothermia-induced neuroprotection.Methods Forty-five SD rats were allocated into mild hypothermia group,TBI group and sham operation group with 15 rats per group according to the random number table.Left-side fluid percussion impact was performed to induce models of TBI.Rats were exposed to hypothermia environment (32-35℃) for 6 hours in mild hypothermia group after TBI.Rats in sham operation group were treated by only drilling on left side of the head,rather than hitting.To evaluate function outcome,modified neurological severity score (mNSS),SEP and synaptophysin mRNA level were measured at 6 hours,24 hours and 7 days postinjury.Results The mNSS in mild hypothermia group lowered compared with TBI group,especially at 24 hours and 7 days (P ＜ 0.05).SEP in mild hypothermia group was significantly shortened at 6 and 24 hours compared with TBI group (P ＜ 0.05),but SEP revealed no significant difference among the 3 groups at 7 days (P ＞ 0.05).Level of synaptophysin mRNA in mild hypothermia group increased at 6 hours postinjury compared with TBI group [(0.08 ± 0.02) vs (0.12 ±0.04)],with further increase at 7 days postinjury[(0.06 ± 0.01) vs (0.33 ± 0.10)] (P ＜0.05).Conclusion The shortage of nerve conduction time of the injured side and promotion of nerve regeneration suggest the neuroprotective role of mild hypothermia following TBI.

Objective To investigate the effect of taurine transporter in the process of protection of brain edema in rats with severe traumatic head injury. Methods A total of 24 Male Sprague-Dawley rats were randomly divided into 4 groups. Except the control rats (Group Sham), all other three groups were subjected to lateral fluid percussion head injury. The TBI (Traumatic brain injury) models (Group TBI) and surgical control rats (Group Sham) were injected with saline through caudal vein after surgery, while the Taurine prevention and Taurine treatment models (Group Pre Tau and Group Tau) were injected with 120 g/L taurine solution before or after surgeries respectively. Water content in each brain, mRNA and protein expres?sion of aquaporin 4 and taurine transporter in the injured rat brain hemispheres were all evaluated over the time course of the study (7 d) in each group. Results Compared with rats in Group Sham, water content in each brain increase, mRNA tran?scription and protein expression of AQP4 were both up regulated but the mRNA transcription and protein expression of TauT were both down-regulated in rats in TBI group. Compared with rats in TBI group, brain water content, mRNA transcription and protein expression of AQP4 all decrease while mRNA transcription and protein expression of TauT all increase in rats in Pre tau and Tau groups. There is no statistical difference of TauT expression between rats in pre-tau group and Tau group. Conclusion Taurine exert its neuron protection role through draining water content from brain and down regulating expres?sion of AQP4 but rising expression of TauT after TBI.

Objective To monitor ventilator-associated event (VAE) for the first time in an intensive care unit (ICU) in China,understand the applicability,incidence,and clinical significance of VAE in China.Methods Targeted monitoring on VAE was performed among patients ≥18 years and with mechanical ventilation (MV)＞2 days in the ICU of a hospital between January 2014 and September 2015,incidence of VAE was calculated,and patients were grouped according to whether or not they had VAE,prognostic factors were analyzed statistically.Results A total of 1 004 patients were monitored,the total hospital stay was 13 795 days in patients who used ventilator,307 (30.58％) cases of VAE occurred,incidence of VAE per 1 000 ventilator-days was 22.25.Univariate analysis showed that patients with VAE had longer length of ICU stay and MV,and higher mortality rate than patients without VAE when they moved out of ICU (all P＜0.05).Multivariate logistic regression analysis showed that VAE was independent risk factor for length of ICU stay,duration of MV,as well as mortality when patients moved out of ICU(all P＜0.05).Conclusion Judgment of VAE is based on MV parameters,it is more objective and accurate.There is a high incidence of VAE among ICU patients,it may lead to poor clinical outcomes,and has good values for the targeted monitoring on ICU patients in large comprehensive hospitals of China.

Objective To investigate effect of taurine on respiration chain enzyme activity of mitochondria 24 hours after severe traumatic brain injury (TBI) in rats.Methods Fifty-six SD rats were divided into sham group,TBI group,taurine treatment group,and taurine prevention group according to the random number table,with 14 rats per group.Fluid percussion brain injury models were used.Via the caudal vein,normal saline was administered to animals in sham and traumatic brain injury groups immediately after injury,while taurine (200 mg/kg)was administered to animals in taurine treatment group after injury and in taurine prevention group 4 days before injury.Brains were harvested 24 hours postinjury for assays of HE staining and electron microscopy.Mitochondrial respiratory chain complex Ⅰ-Ⅴ activities were detected.Results TBI group presented swelling neurocytes,cell loss,karyopyknosis,shortened even vanished process,and inflammation cell infiltration at the edge of necrosis in HE staining.By contrast,morphological improvement was significant in taurine treatment group but only some neurons were intact in taurine prevention group.Swelling mitochondria and broken or vanished mitochondrial crests were seen in TBI group under the electron microscope.However,normal or minor swelling mitochondria was seen in taurine treatment group and cytoplasm slightly porous and absence of mitochondrial crests were seen in taurine prevention group.Activities of complex Ⅰ,Ⅱ and Ⅴ were significant lower in TBI group (32.52±2.41,4.68 ±0.15,2.49 ±0.73) compared to those in sham group (34.03 ±0.46,5.04 ±0.29,3.20±0.68) and in taurine treatment group (33.95±0.85,5.12-±0.23,3.53 ±0.48) (P＜0.05).And complex Ⅰ in taurine prevention group was significantly enhanced as well (34.44 ± 0.36,P ＜ 0.05).Conclusion Taurine may protect the brain tissues and mitochondrial structure from impairment in TBI rats by improving mitochondrial enzymes activity and reducing secondary energy loss.

OBJECTIVEImmortal cell line of human embryonic esophageal epithelium (SHEE) was induced by E6E7 genes of human papillomavirus (HPV) type 18 in our laboratory. To identify the fully malignant transformation at its 85th passage (SHEE85), the malignant phenotype, tumorigenesis and invasive potency were studied.

METHODThe cultured SHEE85 cells were observed under the light and the electron microscope (EM) for cell morphology, analyzed by flow cytometry for cell cycle. The tumorigenesis was assayed by plating cells in soft-agar and transplanting cells into the nude mice and SCID mice. To detect invasive potency, cells were cultured on amniotic membrane in vitro and transplanted into peritoneal cavity of mice in vivo.

RESULTSSHEE85 cells were crowded in cultivation with different sizes and shapes under light microscope, and displayed proliferative morphology under EM. Proliferative index was 47% with 12% hyperploidy cells in determination of DNA histogram. Many large colonies grew in soft-agar (4%) and the transplanted tumors were found in all 4 nude and 4 SCID mice, with strong invasive potency demonstrated in vitro and in vivo.

CONCLUSIONThe immortal esophageal epithelial cell line induced by HPV18 E6 E7 is derived from a fully malignant transformation with a strong invasive potency at the 85th passage. It is also a reliable model for studying the cellular and molecular mechanisms of carcinogenesis of the esophageal carcinoma.

Animals ; Cell Division ; genetics ; Cell Transformation, Neoplastic ; Cell Transformation, Viral ; genetics ; Cells, Cultured ; Epithelial Cells ; cytology ; ultrastructure ; virology ; Esophagus ; cytology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mice, SCID ; Microscopy, Electron ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Ploidies ; Transplantation, Heterologous

OBJECTIVESTo investigate telomerase activity in esophageal squamous cell carcinoma (SCC) and its preneoplasia lesions, and to study the relationships between telomerase activity and cancer differentiation, cancer invasiveness, and lymphatic metastasis.

METHODSTelomerase activity in esophageal SCC tissues, adjacent dysplasia tissues and normal epithelia from the surgical edge were assessed by microdissection-TRAP (telomeric repeat amplification protocol)-silver staining assay.

RESULTSTelomerase activity was detected in 37 (82.2%) of 45 esophageal tumors, 23 (79.3%) of 29 dysplasias, and 2 (5%) of 40 normal epithelia. There was a significant difference in activity between dysplasia and normal epithelium, as well as between tumor and normal epithelium. Twenty-six (92.9%) of 28 tumors with lymphatic metastasis had detectable telomerase activity compared to 11 (64.7%) of 17 non-lymphatic metastasis tumors. These relationships were statistically significant (P < 0.05), but the one between telomerase activity and tumor grade was not.

CONCLUSIONTelomerase activity was high both in esophageal SCC and their preneoplasia lesions. The telomerase activity in SCC tissue was related to lymphatic metastasis, but not to cancer differentiation.

Adult ; Aged ; Carcinoma, Squamous Cell ; enzymology ; pathology ; Cell Differentiation ; Dissection ; Esophageal Neoplasms ; enzymology ; pathology ; Female ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Precancerous Conditions ; Repetitive Sequences, Nucleic Acid ; Telomerase ; genetics ; metabolism ; Telomere

OBJECTIVEStudy on the promoter effects of sodium butyrate in high or low dosages on carcinogenesis process, based on the immortalization of human fetal esophageal epithelium induced by human papillomavirus (HPV) 18E(6)E(7) genes.

METHODSThe immortalized esophageal epithelium SHEE was treated with high concentration of the sodium butyrate (80 mmol/L) and then with low concentration (5 mmol/L) for 8 weeks respectively. The cells were cultured continuously without sodium butyrate for 14 weeks. The morphology, proliferation and apoptosis of the cells were studied by phase contrast microscopy, immunohistochemistry and flow cytometry. The dead and the viable cells were assayed by fluorescent microscopy with Hoechst 33342 and Propidium iodide staining. Tumorigenesis of the cells was assessed by soft agar colony formation and by transplantation of cells into nude mice and SCID mice.

RESULTSWhen cells were exposed to high concentration of sodium butyrate, cell death was increased leaving few live cells. When cells were cultured in the medium with low concentration of sodium butyrate, the first proliferative stage appeared. Removal of the butyrate caused the cell to enter a crisis stage with a long doubling time resembling senescent cells. After the crisis stage, the cells progressed to the second proliferation stage with continuous replication and atypical hyperplasia. At the end of the second proliferative stage, carcinogenesis of the cells appeared with large colonies in soft-agar and tumor formation in transplanted SCID mice and nude mice.

CONCLUSIONSThe malignant change of the immortalized epithelium by the effects of sodium butyrate is the consequence of a two-stage mortality mechanism: cells death by butyrate cytotoxicity and cell crisis by abrogation of sodium butyrate. These data reveal that in high dosage, sodium butyrate induces cell death and in low dosage, it induces cell proliferation, which emphasizes the importance of butyrate as a promotor of carcinogenesis.

Animals ; Butyrates ; toxicity ; Carcinogens ; toxicity ; Cell Death ; drug effects ; Cell Division ; drug effects ; Cell Line, Transformed ; Cell Transformation, Neoplastic ; chemically induced ; Esophageal Neoplasms ; etiology ; Esophagus ; pathology ; Humans ; Mice ; Mice, Inbred BALB C ; Papillomaviridae ; pathogenicity

We analyze the epidemiology,clinical features,and outcome of the patients with Creutzfeldt-Jakob diseases (CJD) in Guizhou Province from 2010 to 2015.The epidemiology,clinical characteristics and follow-up data of CJD suspected patients obtained from Guizhou CJD surveillance network were analyzed.The testing results of cerebrospinal fluid (CFS) and blood from the patients were also collected and analyzed.Results showed that a total of 11 CJD cases was found from 23 reported CJD suspected patients in Guizhou from 2010 to 2015,including 8 probable sporadic CJD(sCJD) cases,2 possible sCJD cases and 1 genetic CJD(gCJD) case.In 11 cases,rapidly progressive dementia was the major initial symptom,following by mental symptoms,extrapyramidal symptoms,signs and cerebellum cortical blindness.Clinical symptoms of progressive dementia were the main symptoms,following by visual or cerebellar dysfunction,myoclonus,cone system/extrapyramidal dysfunction,and akinetic mutism.Most of cases were abnormal in MRI (45.45％) and 14-3-3 protein detection in CSF(70％).The 14-3-3 blood samples of prion gene 129 amino acids (PRNP)polymorphisms were M/M type,excepting for 1 case gCJD confirmed diagnosis cases with D178N mutation in PRNP gene.Eleven CJD cases did not show season and regional clusterings and vocational tendency.The majority of the cases were male,the median age was 65,and mainly were the Han nationality.For all cases of CJD reported during that year for follow-up,the lost-tofollow-up rate was 27％,and the majority of cases died within one year.The sCJD cases were the majority in CJD cases of Guizhou Province,2010-2015.The epidemiological characteristics were similar to the national monitoring cases in the same period.