To investigate the role and mechanism of ceramide (Cer) regulation in alcohol-induced neuronal proliferation and the newborn neurons formation, we used sphingomyelin synthase 2 (predominant enzyme of Cer metabolism) knockout (SMS2(-/-)) and wild type (WT) female mice to establish the model of prenatal alcohol exposure. In 24 h after being given birth (postnatal day 0, P0), the offspring of model mice received blood sphingomyelin (SM) measurement with enzymatic method. On P0, P7, P14 and P30, the proliferation of granule cells in the dentate gyrus and newborn neurons were investigated with immunofluorescent labeling. The expression of protein kinase Cα (PKCα) in the hippocampus was tested with Western blot analysis. The results showed that the SM level of blood in SMS2(-/-) pups was significantly lower than that in WT pups. No matter in SMS2(-/-) or WT mice, the prenatal alcohol exposure down-regulated the SM levels in pups with dose-dependency. In both SMS2(-/-) and WT pups, the number of proliferative neurons and newborn neurons in the dentate gyrus gradually decreased with the growing age. Compared with the WT pups, SMS2(-/-) pups showed significantly more proliferative neurons and newborn neurons in the dentate gyrus. Notably, prenatal alcohol exposure dose-dependently increased proliferative neurons and newborn neurons in the dentate gyrus in both WT and SMS2(-/-) pups. The hippocampal expression of PKCα protein in SMS2(-/-) mice was lower than that in WT mice, and prenatal alcohol exposure could up-regulate the PKCα protein expression in both WT and SMS2(-/-) mice with dose dependency. These results suggest that alcohol exposure during pregnancy can induce the compensatory neural cell proliferation and the production of newborn neurons in offspring, and the Cer-ceramide-1-phosphate (C1P) pathway is involved in alcohol-induced neural cell proliferation. The activation of PKCα may be a key step to start the Cer-C1P pathway and up-regulate the alcohol-induced neural cell proliferation and the newborn neurons formation.
Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Ceramides ; metabolism ; Dentate Gyrus ; cytology ; Ethanol ; toxicity ; Female ; Mice ; Mice, Knockout ; Neurons ; cytology ; Pregnancy ; Prenatal Exposure Delayed Effects ; physiopathology ; Protein Kinase C-alpha ; metabolism ; Signal Transduction ; Transferases (Other Substituted Phosphate Groups) ; genetics

OBJECTIVETo investigate the apoptosis-inducing effect of trichloroethylene (TCE) on cultured normal human epidermis keratinocytes (NHEK) in vitro.

METHODSNR(50) values (the concentration of neutral red absorbed is reduced to 50%) of TCE on NHEK were assayed by neutral red uptake (NRU), and the administered dose of TCE was determined. Lipid peroxidation (LPO) and oxidative stress were assessed by measurement of malondialdehyde (MDA) contens and superoxide dismutase (SOD) activity. Transmission electron microscope (TEM) were used to observe morphologic changes, flow cytometer (FCM) was used to measure DNA contents and calculate cell apoptosis rate and proliferation index (PI).

RESULTSNR(50) values of TCE on NHEK was found to be 4.53 mmol/L (95% CI: 3.92-5.13 mmol/L). The increase in MDA content and inhibition of SOD activity in a concentration-dependent manner were shown after NHEK was treated with a series of dose of TCE 4 h later, and typical morphologic changes of apoptosis were also observed by TEM examination. FCM analysis revealed a sub-G(1) peak in the apoptotic cells. The apoptotic rate in TCE 0.125, 0.500, 2.000 mmol/L exposed groups (31.83%, 38.63%, 44.35%, respectively) were significantly higher than that in blank control (18.42%), while PI in TCE 0.125, 0.500, 2.000 mmol/L group (3.26%, 2.48%, 2.07%, respectively) were significantly lower than that in blank control (4.99%).

CONCLUSIONTCE may induce apoptosis of cultured NHEK in vitro, and inhibit cell proliferation through lipid peroxidation and oxidative stress.

Apoptosis ; drug effects ; Cells, Cultured ; Epidermis ; cytology ; drug effects ; Humans ; Keratinocytes ; drug effects ; Lipid Peroxidation ; Oxidative Stress ; Trichloroethylene ; toxicity

Objective To investigate the relationship between electrolyte level change with prognosis in the patients with craniocerebral injury.Methods A total of 360 patients with craniocerebral injury in this hospital during 2012-2015 were selected as the research subjects and divided into the mild craniocerebral injury group (171 cases),moderate craniocerebral injury group(104 cases)and severe craniocerebral injury group(85 cases) according to the Glasgow coma scale.The severe craniocerebral injury group was further divided into the high level blood sodium subgroup(73 cases)and stable level blood sodium subgroup(12 cases)according to the lev-el of blood sodium,meanwhile 70 persons undergoing healthy physical examination were selected as the control group.The plasma electrolyte levels(blood sodium,potassium,chloride)in each group were detected within 5 d after admission.Then the results were statistically analyzed.Results Compared with the control group,the blood sodium,potassium and chloride levels had no statistical difference between the mild and moderate craniocerebral injury groups(P>0.05).The blood sodium and chloride levels in the severe craniocerebral inju-ry group were higher than those in the mild and moderate craniocerebral injury groups,the difference was sta-tistically significant(P<0.01).The blood potassium level had no statistical difference between the mild,mod-erate and severe craniocerebral injury groups with control group(P>0.05).In the severe craniocerebral injury group,there were 58 cases(79.45%)of death in the high level blood sodium subgroup and 4 cases(33.33%) of death in the stable level blood sodium subgroup,the difference was statistically significant(P<0.01).Con-clusion Clinically monitoring the blood sodium level change in the patients with craniocerebral injury,espe-cially severe craniocerebral injury,is conducive to the disease recovery.

Objective To explore the potential risk factors to Ymman endemic sudden cardiac death (YESCD) and provide evidence for prevention. Methods Twenty-two cases and 24 controls were randomly selected from YESCD areas and non-YESCD areas, respectively. Both cases and controls were interviewed with unified questionnaires. Univatiate X2 test and multivariate conditional logistic stepwise regression was conducted with SPSS 13.0. The optimal regression model was established and evaluated. Results The univariate X2 teat revealed that presence or absence of 5 potential factors might possibly be associated to YESCD: appropriate places for storing dining utensils, pens for livestock, consumption of mushrooms, exposure to any pesticides, and sudden climate changes before onset(X2=12.206,4.779,5.741,6.120,10.754, P<0.05). Multivariate conditional logistic stepwise regression demonstrated that consumption of mushrooms was a protective factor(OR=0.115, P<0.05) and sudden climate change was a risk factor(OR=36.592, P<0.01). Conclusions Sudden climate change might be a risk factor contributing to YESCD, and eating mushrooms before the prevalence seasons may provide some protection.

BACKGROUND AND OBJECTIVECD133-positive colon cancer stem like cells (CSLCs) are resistant to the conventional cytotoxic drug 5-fluorouracil (5-FU). Wnt signaling pathway plays important roles in colon cancer carcinogenesis and metastasis, and regulates the self-renewal capacity of CSLCs. In the present study, we explored the impact of 5-FU on Wnt signaling pathway of CD133-positive colon CSLCs, and the relation between Wnt signaling pathway and drug resistance of CD133-positive colon CSLCs.

METHODSMagnetic activation cell separation was used to collect CD133-positive cells from colon cancer cell line DLD1, which was transfected with luciferase reporter for Wnt signaling activity. The activity of Wnt signaling pathway was compared between CD133-positive and CD133-negative cells. After the treatment with 1 μg/mL of 5-FU, the cell proliferation rates of DLD1 cells, CD133-positive cells, and CD133-negative cells were compared. After the treatment with 1 μg/mL and 10 μg/mL of 5-FU for 48 h, Wnt activity was compared between CD133-positive and CD133-negative cells. The expression of CD133 and cell apoptosis of CD133-positive cells was detected after exposure to 50 ng/mL of dickkopf (DKK)-1, a Wnt pathway inhibitor.

RESULTSAfter the treatment with 5-FU, the cell proliferation rate of CD133-positive cells was higher than that of CD133-negative cells and the sensitivity of CD133-positive cells to 5-FU decreased. Wnt activity was higher in CD133-positive cells than in CD133-negative cells [(46.3 ± 0.3)% vs. (33.9 ± 2.7)%, P = 0.009]. After the treatment with 1 μg/mL and 10 μg/mL of 5-FU, Wnt activity of CD133-positive cells was (90.1 ± 10.0)% (P = 0.012) and (52.9 ± 2.5)% (P = 0.047), respectively, whereas that of CD133-negative cells was (35.5 ± 3.3)% (P = 0.434) and (26.5 ± 0.4)% (P = 0.046), respectively. CD133 expression in CD133-positive cells decreased from (87.2 ± 5.3)% to (60.6 ± 3.1)% (P = 0.022) after treatment with DKK-1, whereas the cell apoptosis rate increased from (11.8 ± 0.2)% to (28.3 ± 0.6)% (P = 0.013).

CONCLUSIONSWnt activity is higher in CD133-positive DLD1 cells than in CD133-negative DLD1 cells. 5-FU can upregulate Wnt activity of CD133-positive colon CSLCs. Blocking Wnt activity may reverse drug sensitivity of CD133-positive cells to 5-FU.

AC133 Antigen ; Antigens, CD ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; metabolism ; pathology ; Drug Resistance, Neoplasm ; Fluorouracil ; pharmacology ; Glycoproteins ; metabolism ; Humans ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Neoplastic Stem Cells ; metabolism ; pathology ; Peptides ; metabolism ; Wnt Signaling Pathway ; drug effects

The aim of this study was to investigate the effect of wogonoside (WGS) on the cisplatin (cDDP) resistance in human gastric carcinoma SGC7901/cDDP cells and its possible mechanism. The drug-resistant SGC7901/cDDP cells were established by stepwise exposure to cDDP. CCK-8 assay was employed to detect the cytotoxic effect of WGS and cDDP on SGC7901/cDDP cells, and the combined effect of WGS and cDDP was analyzed by Chou-Talalay method. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). Nuclear factor erythroid-2 related factor 2 (Nrf2) gene was knocked down by using the short hairpin RNA (shRNA) approach. The protein levels of Nrf2, NAD(P)H:quinone oxidoreductase 1 (NQO1), glutathione S transferase-π (GST-π), multidrug resistance-associated protein 1 (MRP1), cleaved Capase-3, p-Akt and Akt were detected by Western blotting. The result showed that after various concentrations of WGS and/or cDDP treatment for 48 h, the cell viability was remarkably reduced in a dose-dependent manner (P < 0.05). When the inhibition rate exceeded 16%, the combination of WGS and cDDP produced a synergistic effect. The protein levels of p-Akt, Nrf2 and MRP1 in SGC7901/cDDP cells were higher than those in SGC7901 cells (P < 0.05). WGS and LY294002 (PI3K inhibitor) both remarkably decreased the phosphorylation level of Akt (P < 0.05), down-regulated the protein level of Nrf2 (P < 0.05), increased the content of ROS (P < 0.05), up-regulated the protein level of cleaved Caspase-3 (P < 0.05), and induced apoptosis (P < 0.05). Meanwhile, N-acetyl-L-cysteine (NAC) decreased apoptosis and oxidative stress reaction induced by WGS (P < 0.05). WGS and Nrf2 gene silencing both down-regulated the protein levels of NQO1, GST-π and MRP1 (P < 0.05). These results suggest that WGS may reverse cDDP resistance in SGC7901/cDDP cells through blocking the PI3K/Akt/Nrf2/ARE signaling pathway, thus enhancing the cytotoxicity of cDDP and inducing oxidative stress reaction and apoptosis.

OBJECTIVES: To study the association between infliximab trough level (IFX-TL) prior to maintenance treatment and disease outcome in children with Crohn's disease (CD). METHODS: A retrospective analysis was performed on 35 children with CD who received induction therapy with infliximab (IFX) and the measurement of IFX-TL before maintenance treatment from August 2018 to November 2021. Clinical data and laboratory markers at baseline and before maintenance treatment were collected, and the association between outcome and IFX-TL was analyzed. RESULTS: The clinical remission group, endoscopic remission group, and combined remission group had a significantly higher IFX-TL level than the corresponding non-remission groups (P<0.05), and there was no significant difference in the IFX-TL level between the biological remission and non-biological remission groups (P>0.05). The receiver operating characteristic (ROC) curve showed that IFX-TL had an area under the ROC curve of 0.959 (95%CI: 0.894-1) in predicting clinical remission, with a sensitivity of 90% and a specificity of 100% at the optimal cutoff value of 2.3 µg/mL (P<0.001). CONCLUSIONS Among children with CD receiving infliximab induction therapy, the children achieving clinical and endoscopic remission before maintenance treatment tend to have a higher level of IFX-TL. IFX-TL has a certain predictive value for clinical remission.
Child ; Humans ; Infliximab/therapeutic use* ; Crohn Disease/drug therapy* ; Gastrointestinal Agents/therapeutic use* ; Retrospective Studies ; C-Reactive Protein/analysis*

OBJECTIVETo explore the relationship between KRAS gene status and efficacy of Cetuximab (C225) combined with chemotherapy on advanced colorectal cancer in Chinese patients, and to evaluate the safety of C225.

METHODSFrom May 2006 to March 2009, 81 patients with advanced colorectal cancer received Cetuximab combined with chemotherapy were enrolled in this study. The rate of KRAS mutation and the relationship of KRAS with response rate (RR), progression-free survivor (PFS), overall survival (OS) and adverse reaction of C225 were analyzed retrospectively.

RESULTSAll the 81 patients received C225 therapy, and the overall RR was 33.3%. The RR of initiate therapy was 57.1%; of the second line and over the third line therapy was 38.5% and 22.0% respectively. KRAS gene phenotype examination was performed in 44 patients whose tumor samples were available. KRAS mutation was found in 20 cases (45%). Out of 44 patients, 43 were evaluable for response. RR was 5% and 43.48% in KRAS mutation and wild KRAS patients respectively (P =0.002). The median PFS was 7.0 weeks and 18.6 weeks in mutational KRAS patients and wild KRAS patients, reaching statistical significance (P =0.003). The median OS was 15.2 months and 17.3 months in mutational KRAS patients and wild KRAS patients respectively without statistical significance (P =0.463). The common adverse reactions were leucopenia, nausea, vomiting and rash. All the adverse reactions were tolerated. The incidence of skin rash in patients with mutational KRAS and patients without KRAS mutation was 40% and 42% respectively, without statistical significance (P =0.91).

CONCLUSIONC225 combined with chemotherapy is well-tolerated in Chinese patients with advanced colorectal cancer, and it can significantly prolong PFS of patients with wild KRAS as compared to patients with KRAS mutation.

Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cetuximab ; Colorectal Neoplasms ; drug therapy ; genetics ; pathology ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Prognosis ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins p21(ras) ; Retrospective Studies ; ras Proteins ; genetics

OBJECTIVETo understand the genetic polymorphism of DC-SIGN's and DC-SIGNR's neck regions in normal Chinese Han population, and to obtain the genetic data of the two loci in Chinese Han population.

METHODSThe genotypes and alleles of repeat sequences of DC-SIGN and DC-SIGNR neck region were typed by PCR, agarose gel electrophoresis and sequencing. Polymorphism information content (PIC) of DC-SIGNR was calculated.

RESULTSDC-SIGN genetic polymorphism was rare. Allele 7 was most and its frequency was 0.9808. 4-, 5-, 6- and 8- alleles were also found, although their frequencies were very low. Caucasians had only 6- and 8- allele mutants; DC-SIGNR genetic polymorphism was high, its PIC was 0.5312, 4-,5-,6-,7-,8-,9- alleles and 16 genotypes were found in normal Chinese Han population. The differences of 6/5,7/4,7/5,7/6,7/7,9/5,9/7,9/9 genotypes distribution and 5-,6-,7-,9- alleles frequency between normal Chinese Han population and Caucasian population were all extremely distinct (P<0.01). The inserted mutation seemed more in Chinese Hans than Caucasian population.

CONCLUSIONDC-SIGN and DC-SIGNR genotypes and alleles distribution in Chinese Han population are significantly different from Caucasian population and with Chinese own population genetic characteristics, compared with Caucasians.

Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Cell Adhesion Molecules ; genetics ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Lectins, C-Type ; genetics ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Receptors, Cell Surface ; genetics ; Young Adult

This study aims to explore the chemical composition of Rehmanniae Radix braised with mild fire and compare the effect of processing method on the chemical composition of Rehmanniae Radix. To be specific, ultra-high performance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) was used to screen the chemical constituents of Rehmanniae Radix. The chemical constituents were identified based on the relative molecular weight and fragment ions, literature information, and Human Metabolome Database(HMDB). The ion peak area ratio of each component before and after processing was used as the index for the variation. SIMCA was employed to establish principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) models of different processed products. According to the PCA plot, OPLS-DA plot, and VIP value, the differential components before and after the processing were screened out. The changes of the content of differential components with the processing method were analyzed. A total of 66 chemical components were identified: 57 of raw Rehmanniae Radix, 55 of steamed Rehmanniae Radix, 55 of wine-stewed Rehmanniae Radix, 51 of repeatedly steamed and sundried Rehmanniae Radix Praeparata, 62 of traditional bran-braised Rehmanniae Radix, and 63 of electric pot-braised Rehmanniae Radix. Among them, the 9 flavonoids of braised Rehmanniae Radix were from Citri Reticulatae Pericarpium. PCA suggested significant differences in the chemical composition of Rehmanniae Radix Praeparata prepared with different processing methods. OPLS-DA screened out 32 chemical components with VIP value >1 as the main differential components. Among the differential components, 9 were unique to braised Rehmanniae Radix(traditional bran-braised, electric pot-braised) and the degradation rate of the rest in braised(traditional bran-braised, electric pot-braised) or repeatedly steamed and sundried Rehmanniae Radix was higher than that in the steamed or wine-stewed products. The results indicated the chemical species and component content of Rehmanniae Radix changed significantly after the processing. The 32 components, such as rehmapicrogenin, martynoside, jionoside D, aeginetic acid, hesperidin, and naringin, were the most important compounds to distinguish different processed products of Rehmanniae Radix. The flavonoids introduced by Citri Reticulatae Pericarpium as excipient may be the important material basis for the effectiveness of braised Rehmanniae Radix compared with other processed products.
Humans ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Plant Extracts/chemistry* ; Rehmannia/chemistry* ; Flavonoids/analysis*