To investigate the induction of apoptosis of mouse colonic adenocarcinoma CT26 cells by recombinant Clostridium difficile toxin B (rTcdB), CT26 cells were exposed to different concentrations of rTcd B. Inhibition of cell proliferation was measured by MTT assay. The activation of Caspase 3 was measured by colorimetric method. Cell morphological analysis and flow cytometry were performed to confirm cell apoptosis. rTcd B inhibited the proliferation of CT26 cells in a timeand dose-dependent manner. Caspase 3 activity in CT26 cells was elevated remarkably after rTcd B exposure for 6 h, 12 h, 18 h or 24 h, as compared with the control group. Morphological changes were observed by fluorescence microscopy. The exposure of rTcd B to CT26 cells induced a timeand dose-dependent apoptotic cell death as determined by flow cytometry analysis. The results showed that recombinant Clostridium difficile toxin B induced apoptosis of CT26 cells.

Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.

OBJECTIVETo investigate the etiology of 1977 patients from northern China with acute (ALF), sub-acute (SALF) or acute-on-chronic liver (ACLF) failures.

METHODThe age, gender, etiology, pathogenesis, and prognosis of the 1977 patients with liver failures were retrospectively analyzed.

RESULTSOf the 1977 cases, the three most common causes of ALF were HEV (33.96%) or HBV (13.21%) infections or those caused by medicines (9.43%). The three predominant causes of SALF were medicines (31.53%), HEV (16.22%) or HBV (9.91%) infections, but those of the ACLF were HBV (90.29%) infection, alcoholic hepatopathy (2.65%), and HBV super infected with HEV (2.26%) infections. 90.09% (1781) patients were infected by hepatotropic viruses. Of these 1781 patients, the most common cause of their liver failures was HBV infection (92.93%). In these HBV infected patients, 77.10% were from 26 to 55 years old. From 2005 to 2007, there were 39 patients with alcoholic liver failure. In the past two years, there were 23 patients with drug induced liver failure. The improvement rate of the 1977 patients after their treatments was 35.56%. The improvement rate of HEV infected liver failure was higher than drug induced liver failure (P less than 0.05); no statistical significance was found between other groups (P more than 0.05).

CONCLUSIONDifferent types of liver failure have different predominant causes. HBV infection is the most common cause in our 1977 patients. In the past two years, the number of drug induced liver failures and alcoholic liver failures have been increasing.

Acute Disease ; Adult ; Chemical and Drug Induced Liver Injury, Chronic ; etiology ; Chronic Disease ; Female ; Hepatitis B ; complications ; Hepatitis E ; complications ; Humans ; Liver Diseases, Alcoholic ; etiology ; Liver Failure ; chemically induced ; classification ; etiology ; virology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

Purpose: Although the medical decision-making process can be overwhelming for some surrogates, there is a lack of understanding regarding their experiences. The objectives of this study were to examine the decision self-efficacy and decisional conflict experienced by surrogates in intensive care units (ICUs) when faced with the decision of whether to reintubate patients with respiratory failure after a planned extubation. In addition, predictors and mediators influencing these decision-making processes were identified. Methods: This study utilized a cross-sectional design to investigate the decision-making processes of 174 surrogates who were faced with the decision of whether to reintubate patients with respiratory failure after a planned extubation in the internal ICU of a medical center between August 2021 and February 2022. Structured questionnaires were administered to collect data on the surrogates' background information, decision self-efficacy, decisional conflict, and positive and negative affect. The patients’ background information was also collected. Univariate and multivariate analyses were performed to model the data. Results: The mean decision self-efficacy score of the surrogates was 82.41 points, and 20.7% surrogates had decisional conflict scores exceeding 37.5 points, suggesting that they faced challenges in the decision-making process. Surrogates' employment status and negative affect significantly predicted their decision self-efficacy. In addition, patients' activities of daily living prior to hospitalization and the decision self-efficacy of the surrogate significantly predicted surrogate decisional conflict. The impact of surrogates’ negative affect on decisional conflict was fully mediated by decision self-efficacy. Conclusions Surrogate decision self-efficacy mediates the relationship between negative affect and decisional conflict. Providing clinical care interventions that focus on enhancing surrogate self-efficacy and reducing negative affect can help alleviate decisional conflict in this population.

OBJECTIVETo study on effect of concentration of catalpol and 5-hydroxy methyl-2-furaldehyde (5-HMF) from Rehmanniae Radix at various processing.

METHODThe Rehmanniae Radix was dried and prepared from the steaming process with 10% ethanol, 50% ethanol at 90 degrees C and 100 degrees C each other. And the changes of catalpol and 5-HMF was determinated. The extraction of 5-HMF and catalpol was sonicated in 30% methanol for 2 h. The analysis of 5-HMF and catalpol was conducted by HPLC with reversed-phase C-18 column and detected under UV 284 nm, 204 nm. Elution was carried out at 1.0 mL min(-1) with 3% acetonitrile.

RESULTFrom this analysis, we found out that the content of catalpol was decreased with the number of processing times, and content of 5-HMF was increased with the number of processing times at various processing. The temperature and concentration of ethanol can effect on content of catalpol and 5-HMF at processing. The Cooked Rehmanniae Radix processed at 100 degrees C, 10% ethanol is best. And the content of 5-HMF processed for more than 7 times was accorded with standard of Korea phamcopoetia.

CONCLUSIONAnalyze the effect of concentration of catalpol and 5-HMF from Rehmanniae Radix at various processing, and provide the foundation for further study.

Chromatography, High Pressure Liquid ; Ethanol ; Furaldehyde ; analogs & derivatives ; analysis ; Glucosides ; analysis ; Hot Temperature ; Iridoid Glucosides ; Iridoids ; analysis ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry ; Technology, Pharmaceutical ; methods

This study was aimed to investigate the effects of rituximab (RTX), a chimeric human anti-CD20 monoclonal antibody, on lymphoma cell injury induced by X ray irradiation. The human Burkitt EBV-infected and moderate radioresistance lymphoma cells (Namalwa) were used in the this study. Cytotoxicity of rituximab combined with X ray irradiation on Namalwa cells was measured by sulforhodamine B (SRB)-staining; the apoptosis of Namalwa cells was detected by flow cytometry with FITC-Annexin V/PI double staining; the morphologic changes of cells were observed under transmission electron microscope (TEM) and the change of intracellular free calcium level ([Ca(2+)]i) in response to irradiation and rituximab was determined by means of the fluorescent dye fluo-3 and confocal microscopy. The results showed that the growth inhibition in Namalwa cells exposed to irradiation was enhanced by treatment with rituximab. Compared with irradiation alone, rituximab combined with irradiation significantly induced the cell apoptosis and a sustained rise of intracellular free calcium ([Ca(2+)]i) level in Namalwa cells; the serial apoptotic appearances of cells could be observed under TEM. It is concluded that rituximab can enhance the sensitivity of lymphoma cells on X ray irradiation as to induce cell more apoptosis, in this process the intracellular free calcium ([Ca(2+)]i), as an intracellular signaling molecule probably plays an important role.
Antibodies, Monoclonal, Murine-Derived ; immunology ; pharmacology ; Antigens, CD20 ; immunology ; Apoptosis ; Calcium ; analysis ; Cell Line, Tumor ; Humans ; Lymphoma ; drug therapy ; metabolism ; pathology ; Radiation Tolerance ; drug effects ; Rituximab

OBJECTIVETo study the risk factors in high prevalence district of birth defects (BD) to provide evidence for intervention development.

METHODSQuantitative analysis had been carried out based on the investigation at three counties in Shanxi province. 491 useable questionnaires had been collected with a response rate as 94.6%. Binary logistic regression was performed to analyze the risk factors of BD.

RESULTSData from logistic regression analyses revealed that factors as: the status of family income, knowledge on healthy birth and rearing, behavior and illness during pregnancy were influencing the occurrence of BD, with OR values as 0.535, 3.265, 0.403 and 1.379 respectively. Better family income and knowledge on BD were negatively (P values are 0.000 and 0.001 respectively), while illness during pregnancy and alcohol intake of the husbands were positively correlated (P values are 0.005 and 0.012 respectively) to the occurrence of BD.

CONCLUSIONMeasures as accelerating the development of local economy, providing education on BD knowledge to the couples, changing negative life style and improving the health status would reduce the risk of BD. Preconceptional-periconceptional care seemed to be the new idea for healthy pregnancy and healthy baby.

China ; epidemiology ; Congenital Abnormalities ; economics ; epidemiology ; Female ; Health Behavior ; Humans ; Income ; Male ; Pregnancy ; Pregnancy Complications ; epidemiology ; Risk Factors ; Social Class

BACKGROUNDThe risk of clinical deterioration still exists in the acute phase despite the fact that patients with minor stroke may display less severe symptoms. The impact of this clinical deterioration on long-term outcomes is unknown. We characterized the clinical features of neurological deterioration (ND) in the acute phase of minor ischemic stroke (MIS) and investigated its impact on mid- and long-term outcomes.

METHODSThis was a multi-centered, prospective clinical study involving patients with MIS (the National Institutes of Health Stroke Scale, NIHSS ≤3) recruited from the China National Stroke Registry. Patients were included who had been hospitalized within 24 hours of stroke onset. Baseline characteristics, complication rates during hospitalization, etiology of stroke, as well as 3-, 6-, and 12-month post-stroke outcomes were compared between patients with and without ND during the acute phase.

RESULTSA number of 368 (15.2%) out of 2424 patients included in the study exhibited ND in the acute phase. Compared to patients without ND, patients with ND had longer hospital stay, increased rate of baseline diabetes, and multiple complications. Multivariate Logistic regression indicated that ND in acute phase was an independent factor predictive of increased dependence (adjusted odds ratio = 5.20, 95% CI, 3.51-7.70, P < 0.001) at 12-month post-stroke.

CONCLUSIONSThe risk of ND in the acute phase is high in patients with MIS. ND in the acute phase is an independent predictor for poor outcomes at 12 months post-stroke onset.

Aged ; China ; Female ; Humans ; Male ; Middle Aged ; Nervous System Diseases ; etiology ; physiopathology ; Prognosis ; Prospective Studies ; Risk Factors ; Stroke ; complications ; pathology

Objective To investigate the molecular genetic causes and their characteristics of deafness in Ningxia province,we established screening of three common hereditary deafness genes in 336 deaf and hard-of-hearing patients in this district.Methods Peripheral blood samples were obtained from a total of 336 patients with non-syndromic sensorineural hearing loss in parts of special education schools in Ningxia province to extract genomic DNA.The mitochondrial DNA 12S rRNA m.1555A ＞ G mutation was screened by PCR Alw26I digestion and sequence analysis PCR and direct sequencing were used to analyze the coding region of GJB2 and exons 8 and 19 of SLC26A4.Statistical analysis was performed by using SPSS 11.0 software.Frequencies of different GJB2 or SLC26A4 mutations were compared between Han and Hui people.Results Among these 336 patients,seven cases (2.08％,7/336) were found to carry mtDNA 12S rRNA m.1555A ＞ G homozygous mutation,45 cases ( 13.39％ ) were caused by GJB2 mutations and 28 cases (8.33％ ) had two mutated alleles (homozygote and compound heterozygote) of SLC26A4.In detail,16.67％ (56/336) patients carried GJB2 mutations including 11 single mutant carriers.The allele frequency of c.235delC and c.299_300delAT were 9.52％ (64/672) and 2.68％ ( 18/672),respectively,making up 81.19％ (82/101) of all pathogenic mutated alleles for GJB2.The single mutant allele carriers of SLC26A4 is 32,and two types (c.919-2A ＞ G and c.2168A ＞ G) accounted for 95.29％ (24/27)mutations,totally.We also found that statistically significant differences in c.919-2A ＞ G and c.2168A ＞ G frequencies between Han and Hui people ( c.919-2A ＞ G,x2 =8.229,P =0.004 ; c.2168 A ＞ G,x2 =5.277,P =0.022).However,there was no statistically significant difference in GJB2 mutation between Han and Hui people.Conclusions GJB2 mutation was a primary causc for non-syndromic sensorineural hearing loss in Ningxia province,and c.235delC was the most common mutant forms of GJB2.c.919-2A ＞ G and c.2168A ＞ G were common mutant forms of SLC26A4,their frequencies were also statistically significant differences between Han and Hui people.

Background and Objective:Recently,the theory of cancer stem cells(CSCs)has presented new targets and orientations for tumor therapy.The major difficulties in researching CSCs include their isolation and purification.The aim of this study is to identify and characterize the side population(SP)cells in small cell lung cancer(SCLC)cell line H446,which Iays the foundation for the isolation and purification of CSCs.Methods:Fluorescence-activated cell sorting(FACS)was used to sort SP and non-SP (NSP)cells from H446,Both subgroups were cultivated to survey the capacity to form into suspended tumor cell spheres.Reverse transcriptionpolymerase chain reaction(RT-PCR)and real-time PCR were used to evaluate the expression levels of the mRNA of CD133,ABCG2,and nucleostemin in both subgroups.The capacity of proliferation and the differences in drug resistance of both subgroups and unso rted cells were tested by the MTT method.The differentiation ability of both subgroups was determined by FACS.Proliferation was determined by subcutaneous tumor formation in nude mice.Results:The percent of Hoechst 33342 negative cells was about(5.1±0.2)%in H446 by fluorescence microscopy.The percent of SP cells was(6.3±0.1)%by flow cytometry.SP cells had a stronger capability of fOrming into tumOr spheres than NSP cells.The mRNA expression Ievels of ABCG2,CD133,and nucleostemin in SP cells were 21.60±0.26,7.10±0.14,and 1.02±0.08 folds higher than that in NSP cells(P＜0.01,P＜0.01,and P＞0.05,respectively).In vivo,SP cells showed better proliferative ability and tougher viability when treated with drugs.SP cells can differentiate into NSP cells,but NSP cells cannot differentiate into SP cells.SP cells had a greater ability to form tumors.Conclusions:The H446 cell line contained some SP cells with stem cell properties.CD133 and ABCG2 may be cancer stem celI markers of SCLC.