AIM: To study the roles and mechanisms of ERKs and intracellular free calcium in cardiomyocyte hypertrophic response induced by endothelin-1(ET-1). METHODS: (1) Neonatal rat cardiomyocyte hypertrophic response was assayed by measuring cell surface area and protein content; (2) ERKs activity was determined by Whatman Paper Filter method; (3) Intracellular free calcium concentration ([Ca 2+ ]i) was measured using Fura-2/AM as a fluorescent indicator. RESULTS: (1) ET-1 could increase total protein production, surface area, ERKs activity and [Ca 2+ ]i in cultured cardiomyocyte in dose-dependent manner at concentrations ranging from 10 -9 to 10 -7 mol/L. And this effect could be abolished by BQ123, an antagonist of ET A receptor, partly inhibited by PTX, but not by BQ788, an antagonist of ET B receptor.(2)The activation of ERKs and the increase of [Ca 2+ ]i induced by ET-1 were obviously inhibited by PD98059, a selective ERKs kinase inhibitor, and nifedipine, a calcium channel blocker, respectively. Both antagonists partially inhibited ET-1-stimulated cardiomyocyte hypertrophic response. (3) Staurosporine, a selective PKC inhibitor, could inhibit ET-1-stimulated cardiomyocyte hypertrophic response and increase of [Ca 2+ ]i, but not affect the activation of ERKs. CONCLUSION: Cardiomyocyte hypertrophic response induced by ET-1 is mediated by ET A receptor coupled to PTX-sensitive G-protein, which involves at least two signalling pathways: PKC-mediated increase of [Ca 2+ ]i , and PKC-independent activation of ERKs. [

BACKGROUND: During the process of isolating and culturing neural stem cells, trypsin is used to digest tissues and cells, but it is difficult to control the digestion time.OBJECTIVE: To isolate and cultivate neural stem cells from fetal brain of Kunming mice by trypsinization combined with mechanical isolation and to identify by the immunohistochemical method.DESIGN, TIME AND SETTING: In vitro observation of cytology. The experiment was performed at Basic Medical Laboratory of Guangxi Medical University from October 2006 to September 2007.MATERIALS: Kunming mice of pregnant 14 to 16 days were provided by the Laboratory Animal Center of Guangxi Medical University.METHODS: Brain tissues were isolated from the fetal brain of Kunming mice, pipettad mechanically after trypsinization, and incubated in serum-free DMEM/F12 medium containing B27, basic fibroblast growth factor and epidermal growth factor.MAIN OUTCOME MEASURES: Immunohistochemistry was performed to identify the cultured neural stem cells.RESULTS: After incubation in serum-free DMEM/F12 medium for 24 hours, the cells were suspensions with the manner of little neurospheres. After incubation for 48 hours, the cultured cells grew much larger, formed typical neurospheres, with no marked process, and survived well after passaged. In addition, immunocytochemical method showed nestin-positive.

Objective To evaluate the effects of continuous intra-tracheal gas insufflation (TGI) during mechanical ventilation for protecting the juvenile piglets with acute lung injury (ALI) induced by endotoxin. Method Twelve healthy juvenile piglets were anesthetized and mechanically ventilated at 2 cmH2O PEEP with 10 cmH2O peak inspiration pressure. The piglets were challenged with lipopolysaccharide (LPS) and randomly (random number) assigned to two groups (n = 6 each): (1) piglets treated with mechanical ventilation alone (group MV) and (2) piglets treated with TGI by continuous airway flow of 2 L/min (group TGI). FiO2 was set at 0.4 to avoid oxygen toxicity, and the piglets were continuously monitored with an oxygen analyzer. Results Tidal volume, ventilation efficacy index and mean airway pressure were significantly improved in piglets of TGI group (P < 0.01 or P < 0.05). Four hours after ALI, pH decreased to below 7.20 in piglets of MV group, and was higher in piglets of TGI group (P < 0.01). Similarly, PaCO2 was stable and was significantly lower in piglets of TGI group than that in piglets of MV group (P < 0.01). PaO2 and PaO2/FiO2 increased in piglets of TGI group (P < 0.05). There were no significant differences in heart rate, respiraaatory rate, mean arterial pressure, central venous pressure, dynamic lung compliance and mean resistance of airway between two groups. Lung histopathological changes showed severe inflammation,and intra-alveolar hemorrhage and interstitial patchy hemorrhage were ameliorated and the lungs were more homogenously expanded in piglets of TGI group. Conclusions Continuous TGI during MV can significantly improve gas exchange and ventilation efficacy, and may provide a better treatment for acute lung injury.

The development of cholestatic liver disease is closely associated with the enterohepatic circulation disorder of bile acid.This article elaborates on metabolism of bile acid,enterohepatic circulation-related mechanism of hereditary cholestasis,molecular regulation of related transporters,and association between current therapeutic regimens and enterohepatic circulation,in order to investigate the pathogenesis of cholestatic liver disease and provide a basis for the research on new drugs or therapies.

OBJECTIVETo investigate the predominance ticks and the infectious status of severe fever with thrombocytopenia (SFTSV) in Penglai and Laizhou counties, Shandong province.

METHODSTwo towns with high incidence rate were selected in Penglai and Laizhou, respectively, then three villages were selected in each towns. Parasitic ticks were collected from the host skin by hand manually and free ticks manually with white cloth from the grassland, monthly, during April to December in 2011. Samples were classified by original, varieties, developmental stages, then extracted RNA, using Realtime RT-PCR to test severe fever thrombocytopenia syndrome virus, S fragments were amplified with nested PCR, then isolated virus. By neighbor joining method in the phylogenetic tree, the minimum infection rate (MIR) was used to represent the infection status of ticks in novel bunyavirus.

RESULTSA total of 3 145 ticks were collected totally from 5 categories, there were 3 048(96.92%) of Haemaphysalis longicornis, 73(2.32%) of Rhinpicephalus sanguineus, 10(0.32%) of microplus Boophilus, 9(0.29%) of Haemaphysalis campanulata, 5(0.16%) of Dermacentor sinicus, respectively. The positive rate of nucleic acid of 2 044 samples was 6.16% (126/2 044), minimum infection rate (MIR) was 4.01%, there were 122(96.83%) of Haemaphysalis longicornis, 3(2.38%) of Rhinpicephalus sanguineus, and 1(0.79%) of microplus Boophilus, MIR was 4.00%, 4.11%, and 10.00%, respectively. There were no nucleic acid positive samples in Haemaphysalis campanulata and Dermacentor sinicus. The 11 S segments were amplified in 126 positive samples, the homology of S fragment was 95.6%-99.9% with 11 strains isolated from the identified SFTS cases in local area, 3 strains isolated from animals, and 11 strains isolated from other areas. There was no significant difference among original, varieties and developmental stages.

CONCLUSIONHaemaphysalis longicornis was the predominant species in Penglai and Laizhou counties, it could be propagation medium with Rhipicephalus sanguineus and microplus Boophilus, S sequence in ticks was higher homology with virus isolated from local SFTS cases.

Objective To establish and evaluate the predictive value of the risk prediction model for lung infection within postoperative 1 year in kidney transplant recipients. Methods Clinical data of 197 kidney transplant recipients were retrospectively analyzed. All recipients were divided into the infection group (n=42) and non-infection group (n=155) according to the incidence of lung infection within postoperative 1 year. The incidence and risk factors of lung infection after kidney transplantation were analyzed. Risk prediction model was established by multiple logistic regression analysis. Forty-five kidney transplant recipients who met the inclusion criteria, including 8 cases in the infection group and 37 cases in the non-infection group, were selected to verify the predictive effect of the established model. Results The incidence of lung infection within 1 year after kidney transplantation was 21.3% (n=42), including 38 cases (90%) of pneumonia severity index (PSI) class Ⅰ, 1 case (2%) of PSI class Ⅲ and 3 cases (8%) of PSI class Ⅴ. Lung infection occurred within 1 month after operation in 13 cases, within postoperative 2-6 months in 22 cases and after postoperative 6 months in 7 cases. Nineteen recipients were diagnosed with bacterial infection, 7 cases of fungal infection, 10 cases of viral infection and 6 cases of mixed infection. Smoking history, diabetes mellitus history, pulmonary disease history and albumin level of < 35 g/L were the independent risk factors for lung infection after kidney transplantation (all P < 0.05). The equation of risk prediction model for postoperative lung infection in kidney transplant recipients was logit (lung infection within postoperative 1 year in kidney transplant recipients)=-1.891+1.063×smoking history (yes=1, no=0)+1.398×diabetes mellitus history (yes=1, no=0)+1.732×pulmonary disease history (yes=1, no=0)+1.269×albumin level (< 35 g/L=1, ≥35 g/L=0). The area under the curve (AUC) of receiver operating characteristic (ROC) was 0.788, the sensitivity was 0.786, the specificity was 0.645, and the Youden index was 0.431, respectively. Hosmer-Lemeshow goodness-of-fit test demonstrated that the predicted value of this model yielded relatively high consistency with the observed value. The AUC in the verification group was 0.834. Hosmer-Lemeshow goodness-of-fit test validated high degree of calibration of this model. Conclusions The risk prediction model, consisting of smoking history, diabetes mellitus history, pulmonary disease history and albumin level as predictors, may effectively predict the incidence of lung infection within postoperative 1 year in kidney transplant recipients.

Objective: To explore the relationship between genotype and phenotype of ABCB11 deficiency. Methods: Clinical data of two siblings with ABCB11 deficiency were retrospectively analyzed. Related literature from PubMed, CNKI and Wangfang databases was reviewed to date (up to August 2017) with 'ABCB11 gene’ or 'bile salt export pump’, 'cholestasis’ and 'child’ as key words. Results: The patients were siblings. Both of them presented as jaundice, pruritus and hepatosplenomegaly since 3 days after birth. Significant laboratory findings on admission of the older sister included high total bilirubin, 170 µmol/L;conjugated bilirubin, 115.8 µmol/L;alanine aminotransferase, 168 U/L;total bile acid 186.3 µmol/L and normal gamma-glutamyl transpeptidase. While routine laboratory data of the younger brother were as follows: total bilirubin, 148.8 µmol/L;conjugated bilirubin, 96.3 µmol/L;alanine aminotransferase, 232.8 U/L;total bile acid 226 µmol/L, and normal gamma-glutamyl transpeptidase.Both received ursodeoxycholic acid and fat-soluble vitamins. Liver pathology of the younger brother showed giant hepatocytes with ballooning degeneration, focal necrosis and intrahepatic cholestasis. Both the patients harbor the same compound heterozygous mutations in ABCB11 gene, c.145C>T (p.Q49X) and c.1510G>A (p.E504K). The sister is 9 years old now, with normal liver function. Jaundice faded around 3 months after birth, pruritus relieved at age 5, and medications was stopped since then. The brother progressed to liver failure after an operation on perianal abscess when he was 8-month-old, and received living-related liver transplantation when he was 9 month and 20 days old (from his mother). Now he is 1 year and 5 months old, with normal liver function. Both are under our follow-up. Literature review revealed 18 ABCB11 deficiency patients from 7 families who had apparent different prognoses, within each family the siblings had the same ABCB11 gene mutation. Seven cases relieved after ursodeoxycholic acid therapy and/or partial external biliary diversion, 5 received orthotopic liver transplantation, 2 developed hepatocellular carcinoma and 4 cases died in childhood. Conclusions The clinical manifestations of ABCB11 deficiency may vary greatly in patients carrying the same genotype, even in siblings. Patients should be managed in individualized maner.

Objective To learn the prevalence of infection of human and animals severe fever with thrombocytopenia syndrome bunyavirus(SFTSV)in Yantai,Shandong province,and to analyze the pathogenic features of SFTSV as well as its relationship between human and animal hosts. Methods From April to November in 2011,3 576 serum samples were collected from domesticated animals,including sheep,cattle,pigs,dogs,chickens,in Laizhou and Penglai areas where fever with thrombocytopenia syndrome frequently occured among local residents. Total SFTSV antibodies and virus-specific nucleic acids of the serum were tested by ELISA and Real time RT-PCR,respectively. SFTSV infection on each animal was observed in different months. 2 590 human serum samples were also collected in Laizhou and Penglai areas,with IgG antibodies tested by ELISA. Virus was isolated with Vero cells from the serum which SFTSV viral nucleic acids were positive. S fragments were amplified by RT-PCR and sequenced,with homology analysis conducted on these sequences. Results The overall positive rate of serum samples from animals on the total SFTSV antibodies was 40.24%(1 439/3 576)while the positive rate for specific nucleic acids was 4.56%(163/3 576). The positive rates for SFTSV antibodies were 62.78%,52.97%,45.56%,28.73%,1.45%and the positive rates for specific nucleic acids were 5.72%,4.63%,3.02%,5.25%and 3.73%,in sheep,cattle,chickens,dogs, pigs,respectively. The antigens/antibodies for SFTSV in animals changed seasonally. The overall positive rate for SFTSV IgG antibody from 2 590 human samples was 5.41%. Thirteen virus strains were isolated from these serum samples(10 strains from human and 3 strains from animals). The nucleotide homology of 13S fragments’sequences ranged from 95.23%to 100.00%and the nucleotide homology with the isolates from other provinces were between 94.72%and 99.13%. The homology was considered to be high. Conclusion High prevalence of SFTSV infections occured both in human and domestic animals in Yantai city. The nucleotide sequences of SFTSV were highly homologous among human and domestic animals. The findings suggested that domesticated animals might serve as SFTSV proliferation and the hosts for transmission thus should be attached great importance.

Objective To observe the clinical characteristics and fundus imaging features of unilateral acute idiopathic maculopathy (UAIM).Methods Retrospective cases series.Six eyes of 6 patients with UAIM were included in this study.There were 4 males and 2 females.All patients underwent best corrected visual acuity (BCVA),fundus color photography,spectral domain optical coherence tomography (SD-OCT) and fundus fluorescein angiography (FFA) examinations.3 patients underwent indocyanine green angiography (ICGA) examination.No intervention was given in all patients after diagnosis.The mean follow-up was 6.3 months.BCVA,fundus color photography,SD-OCT and FFA were performed in follow-up.Results The ocular symptoms included unilateral sudden decreased vision,metamorphopsia and central scotoma.All patients had a yellow-white lesion involving the macular and irregular small bleeding was seen in 3 eyes.Irregular hyperfluorescence in macular area was seen in all patients,some with the hemorrhages showed blocked fluorescence at the early stage of FFA and fluorescence storage at the late stage.ICGA showed that hypo-fluorescence in macular area continually.SD-OCT showed that subretinal fluid in the macular and lost the ellipsoid zone.During the follow-up period,the subretinal fluid was absorbed spontaneously in one month.The ellipsoid zone structure recovered gradually and the visual acuity were normal.At the time of the latest follow-up,the BCVA was 0.8 in 4 eyes,1.0 in 2 eyes.Conclusions UAIM is featured with a unilateral,acute,transient and exudative lesion in the macular,which can be spontaneously resolved;It mainly affects outer retina.

OBJECTIV E To study composition an d content changes of volatile components during the bleaching process of Atractylodis macrocephala with the water of washing rice. METHODS The raw products of A. macrocephala and bleached products of 5 different bleaching stages were prepared （in the first and second stages ，raw products were bleached with 9-fold volumn of the water of washing rice for 12 h and 24 h，respectively；in the third ，fourth and fifth stages ，the raw products were firstly bleached with 9-fold volumn of the water of washing rice for 24 h，and then bleached with 9-fold volumn of clean water for 12，24 and 48 h，respectively）；the bleaching temperature was set at 26 ℃. The volatile components of raw products of A. macrocephala and its bleached products of 5 different bleaching stages were qualitatively analyzed by using headspace gas chromatography-mass spectrometry. The relative percentage of each component was calculated by peak area normalization method. RESULTS A total of 49 volatile components were identified from raw products of A. macrocephala and its bleached products of 5 different bleaching stages，including 20 common volatile components such as terpinolene ，cyperene and atractylon ，etc. Among them ，33，31，28， 30，28 and 29 volatile components were identified from the raw products of A. macrocephala and the bleached products of the first to fifth stages ，the relative percentages of which were 66.218％ ，64.711％ ，79.410％ ，65.419％ ，67.101％ ，66.818％ ， respectively；among them ，the relative percentage of atractylon in bleached products was the highest in the fourth stage （41.206％），but was the lowest in the third stage （35.926％）. Compared with the raw product ，16 volatile components such as pethylbrene and β-vetivenen were added in the bleaching process ，while 8 volatile components such as ethyl palmitate and β-maaliene were not detected. However ，5 volatile components including 11-rotundene and （－）-valeranone in the bleaching process showed a trend of disappearance-emergence and disappearance-emergence-disappearance. CONCLUSIONS In the third stage，the total relative percentage of each volatile component and the relative percentage of representative dry component as ， atractylone are the lowest in bleached products of A. ； macrocephala，i.e. the bleaching technology of relieving the dry property of A. macrocephala e with the water of washing rice is bleaching with 9-fold volumn of the water of washing rice for 24 h，and then bleaching with 9-fold volumn of clean water for 12 h.