The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.

Animals ; Hepatic Stellate Cells ; Humans ; Liver Cirrhosis ; therapy ; Liver Regeneration ; Mesenchymal Stromal Cells

Objective?To evaluate the effect on removal of ovarian cyst by laparoendoscopic single site surgery and enhanced recovery after surgery (ERAS).?Methods?A prospective, single-institution study was performed for patients who were diagnosed benign ovarian cyst, underwent removal ovarian cyst, and adopted ERAS nursing care from June 2015 to June 2017. 40 patients who adopted laparo-endoscopic single site surgery were experimental group and 40 patients who adopted traditional laparoscopy surgery were control group. We compared the operation time, blood loss volume during operation, the time of getting out-of bed after operation, the postoperative exhausting time, the defecation time after surgery, the incidence of postoperative febrile and other complications, the time of hospital stay, and hospitalization expenses between the two groups. The measurement data was tested by t test, and the counting data was tested by χ2 test, which was statistically significant with P < 0.05.?Results?The results showed that the time of getting out-of bed after operation, the postoperative exhausting time, the defecation time after surgery and the time of hospital stay in experimental group was significantly shorter than the control group;Meanwhile the hospitalization expense was lower than the control group. These results were statistically significant (P < 0.05). While there was not statistically significant in the operation time, blood loss volume during operation, and the incidence of postoperative febrile and other complications between the experimental group and the control group (P > 0.05) .?Conclusion?ERAS combined with laparo-endoscopic single site surgery is helpful to the reduction of hospitalization cost and the clinical promotion and application.

OBJECTIVETo study the effect of Shenfu Injection (SF) on the apoptosis of prostate cancer PC-3 cells and its possible mechanism.

METHODSWe divided prostate cancer PC-3 cells into a blank control group and three experimental groups, the latter treated with SF at 50, 100, and 200 microl/ml, respectively, for 24, 48, and 72 hours. Then we determined the proliferation of the cells by MTT assay, measured their apoptosis by Annexin V/PI flow cytometry, and detected the expression of P53 mRNA by RT-qPCR.

RESULTSCompared with the blank control group, the survival rates of the prostate cancer PC-3 cells in the 50, 100, and 200 microl/ml SF groups were (93.76 +/- 2.63)%, (81.21 +/- 1.80)% and (18.01 +/- 3.84)% at 24 hours, (94.67 +/-1.11)%, (78.33 +/- 2.89)% and (10.34 +/- 1.44)% at48 hours, and (91.30 +/- 0.47)%, (36.67 +/- 1.56)% and (1.33 +/- 0.32)% at 72 hours, all significantly increased in a dose- and time-dependent manner (P < 0.05). The expression of p53 mRNA was also markedly increased in all the three experimental groups at 48 hours (P < 0. 05).

CONCLUSIONSF can inhibit the proliferation and induce the apoptosis of PC-3 cells, which may due to its upregulation of the p53 mRNA expression.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

Objective: To explore the clinical characteristics and prognostic factors of the patients with non-Hodgkin's Lymphoma (NHL) complicated with HBV infection, so as to provide a basis for clinical accurate diagnosis and prognosis evaluation. Methods: The data of 313 newly diagnosed NHL patients from August 2012 to July 2016 were collected. The HBV serological markers were detected by ELISA, and HBV DNA was quantified by full automatic microparticle chemiluminescence immunoassay (≥1×10(5) copies/ml as high copy group, 1×10(3)-<1×10(5) copies/ml as low copy group). The relationship between HBV infection and prognosis was analyzed combined with the clinical features of the patients, and the HBV detection rate was compared with that of the common population (from the national HBV sero epidemiological data). Results: ①The positive rate of HBsAg in NHL patients was 12.5% (39/313), which was higher than 7.2% in the general population (χ(2)=14.596, P<0.001). HBV infection in the past (HBsAg negative but HBcAb positive) in 114 cases (36.4%), the incidence was slightly higher than that in the general population (34.1%). ②Compared HBsAg positive group with the negative group, the proportion of B cell type (87.2% vs 70.3%, P=0.027), Ann Arbor stage Ⅲ-Ⅳ(69.2% vs 34.6%, P<0.001), IPI score 3-5 (74.4% vs 50%, P=0.004), LDH level (79.5% vs 47.8%, P<0.001) and liver involvement (45.5% vs 31.7%, P=0.006) were all higher. The difference was statistically significant. ③Compared the HBV infected group (114 cases) with the non-infected group (160 cases), the difference had statistical significance in the proportion of Ann Arbor stage Ⅲ-Ⅳ (P=0.023) and IPI score 3-5 scores P=0.035). ④Compared HBV DNA positive group (30 cases) with negative group (71 cases), Ann Arbor stage Ⅲ-Ⅳ (P=0.011), IPI score 3-5 score (P=0.030), LDH level (P=0.025) and liver involvement (P<0.001) in the proportion of patients had statistical significance. The positive patients were divided into HBV DNA high and low copy groups with 1×10(5) copies of /ml as the boundary. The results showed that there was no statistical difference between the two groups (P>0.05). Conclusions: The HBV infection rate of NHL patients is significantly higher than that of the general population, and HBV infection is more closely related to B cell type NHL. Patients with HBV infection and HBV DNA positive had late Ann Arbor stage, high IPI score, high LDH level and liver involvement, and the prognosis is poor.
Hepatitis B ; Hepatitis B Antibodies ; Hepatitis B Surface Antigens ; Humans ; Lymphoma, Non-Hodgkin ; Prognosis

Objective To describe the epidemiologieal features of viral encephalitis and burden of Japanese encephalitis (JE),and to identify potential strategies for effective JE control measures,using data from the Viral Encephalitis Surveillance Program (VESP) launched in Ankang,Baoji,and Weinan prefectures,Shaanxi province.Methods Data was gathered from sentinel hospitals reporting system on all the viral encephalitis (VE) eases identified between June 2005 and May 2007.County Center for Disease Control and Prevention (CDC) investigated the cases,drawing blood and cerebrospinal fluid (CSF) samples from the hospitals,and testing IgM antibody against JE using ELISA.We used Epi Data and Excel for data entry and analysis.Results A total of 1097 VEs were reported and 1053 (96.0%) had blood or CSF samples collected and tested for IgM antibody against JE.Three hundred and eleven cases (29.5%) showed JE antibody positive (JE confirmed case).Among the JE confirmed cases,numbers of those under 15 year of age accounted for 33.7%,43.9%and 88.3%in Baoji,Weinan and Ankang prefectures respectively.The rest were mainly children aged 5-14 years old (53.3%).Toddlers,farmers and children accounted for 85.2%in JE confirmed cases.About half of other VE cases (51.0%) were students of all age.Data an investigation on 398 reported VE cases at discharge,showed that 67.1%of JE confirmed cases recovered while 83.7%of the other VE cases fully recovered.The case fatality rates were 9.2%for JE confirmed cases and 3.1%for other VE cases.578 cases were followed up at 90-days after discharge,69.6%of JE confirmed cases and 90.2%of other VE cases recovered,with case fatality rates were 13.6%and 3.6%for JE confirmed cases and for other VE cases,respectively.The sequelae rates were 10.0%for JE confirmed cases and 4.5%for other VE cases.Conclusion The peak of the VE season was the sameas that of JE.There were 45.6%of reported JE cases with negative JE IgM,suggesting that it is necessary to carry out laboratory testing for clinical diagnosis cases.The fact that high risk population was different at prefectures levels suggested that more attention be paid in JE control and prevention.

OBJECTIVETo investigate the value of the HCT-CI score in chemotherapy risk assessment and prognosis of elderly patients with acute myeloid leukemia (AML).

METHODSThe clinical data of 116 AML patients older than 60 years in the department of Hematology, Henan Provincial People's Hospital from January 2000 to December 2010 were analyzed retrospectively. All patients received cytarabine-based regimens, including protocol DA, MA, IA, AA or CAG, followed by cytarabine-based postremission treatment. (1) Comorbidities were evaluated by using HCT-CI score, the early death rates and median survival time were compared among these different groups. (2) These prognostic factors were analyzed by univariate and multivariate analyses.

RESULTS(1) All 116 cases were followed-up. The patient cohort was divided into those with HCT-CI scores of 0, 1 or 2, or ≥ 3. Early death rates were 3.7%, 12.1% and 23.21% in above three groups, respectively (P < 0.01). Overall survival were 345, 225 and 113 days, respectively (P < 0.01). (2) HCT-CI score ≥ 3 (P < 0.01), antecedent MDS history (P = 0.035), high-risk karyotype (P = 0.018), white blood cells at diagnosis ≥ 100×10(9)/L (P = 0.041) were independent adverse prognostic factors with multivariate analysis.

CONCLUSION(1) The HCT-CI score can objectively assess elderly AML patients with comorbidities and predict chemotherapy risk in older patients receiving AML induction therapy. (2) Antecedent MDS history, high-risk karyotype, high white blood cell, and HCT-CI score ≥ 3 are independent adverse prognostic factors of elderly AML patients.

Aged ; Aged, 80 and over ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Assessment ; Treatment Outcome

OBJECTIVETo investigate the effects of rhubarb powder on serum complement 3 (C3), complement 4 (C4), and hypersensitive C-reactive protein (hs-CRP) levels in patients with hypertensive intracerebral hemorrhage (HICH) after operation.

METHODSForty inpatients with HICH after operation were recruited from Department of Cerebral Surgery, Affiliated Hospital of Shaanxi College of Traditional Chinese Medicine from July 2009 to March 2010. They were randomly assigned to the treatment group (20 cases) and the control group (20 cases). From the 4th day after surgery, all patients received routine Western medical treatment. The rhubarb powder, 5-10 g dissolving in 40 mL warm water, was administered or nasally fed to those in the treatment group, 2 -3 times daily for 10 successive days. The contents of serum C3, C4, and hs-CRP were detected in the two groups on the 7th day and the 14th day after operation. The serum hs-CRP content was detected using latex particle enhanced immunoturbidimetric assay. The Scandinavia Stroke Scale (SSS) scores were recorded in the two groups.

RESULTSCompared with the same group on the 4th day after operation, the levels of serum C3 and C4 increased on the 7th day after operation, and SSS score increased on the 14th day after operation in the control group (P < 0.05). The contents of C4 and hs-CRP decreased, and the SSS score increased on the 14th day after operation in the treatment group (P < 0.05). Compared with the same group on the 7th day after operation, the contents of C4 and hs-CRP decreased and the SSS score increased on the 14th day after operation in the treatment group (P < 0.05). Compared with the control group at the same time points, the contents of C4 and C3 decreased on the 7th day after operation; the contents of C3, C4, and hs-CRP decreased, and SSS score increased in the treatment group on the 14th day after operation (P < 0.05).

CONCLUSIONThe rhubarb powder could significantly decrease the serum levels of C3, C4, and hs-CRP, and improve the curative effect in patients with HICH after operation.

Adult ; C-Reactive Protein ; metabolism ; Cerebral Hemorrhage ; blood ; Complement C3 ; metabolism ; Complement C4 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Postoperative Period ; Rheum ; chemistry

OBJECTIVE: To investigate the clinical and pathologic features of melanocytic lesion of the central nervous system. METHODS: We analyzed the clinical features, neuroimages, and operational and neuropathological findings of 2 patients of neurocutaneous melanosis and 4 patients of primary leptomeningeal melanoma. RESULTS: All the 6 patients had the common clinical features of intracranial hypertension and epilepsy. Brain CT and MRI showed abnormal signals. More melanin pigment nevi were found on the skin of the 2 patients subjected to neurocutaneous melanosis. The other 4 patients subjected to primary leptomeningeal melanoma had no melanin pigment nevus on the skin, but the brain section displayed positive meningeal melanoma. CONCLUSION Intracranial hypertension and epilepsy are the main clinical manifestations of melanocytic lesion of the central nervous system, and cutaneous lesion and radiological findings are very important for the diagnosis.
Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Melanosis ; pathology ; Meningeal Neoplasms ; pathology ; Middle Aged ; Nervous System Diseases ; pathology ; Neurocutaneous Syndromes ; pathology ; Young Adult