Objective To observe the effect of astragaloside IV (A) and SB203580 antagonize cadmium (Cd) toxicity on expression of associated protein and blood-testis barrier(BTB) in rats and to study the protective mechanism of A on it.Methods Totally 21 SD male rats were randomly divided into 7 groups, 3 rats per group:Cd [ intraperitoneally injected with 0.1%CdCl2,1mg/(kg?d)],Cd+A [at the above dose of CdCl2,at the same time with A,10mg/(kg?d)], and Cd +SB203580 [at the above dose of CdCl2,at the same time with SB203580,100μg/(kg?d)], each of the above groups was further divided into continuous five and ten days treatment groups .The control group was intraperitoneally injected with equal dosage of normal saline .The testes were studied by light , electron microscopy , immunohistochemistry and Western blotting .Results In the control group ,irregular and lightly stained nuclei of Sertoli cell ( Sc) in seminiferous tubules were observed by HE staining .A continuous electron density line of tight junction ( TJ) and normal ultrastructure of BTB were observed .After Cd treatment ,the vesicular formation in the Sc was observed .The ultrastructural damage of Sc and TJ was observed .Compared with the corresponding time point of Cd group ,these were weakened in morphology of testis and ultrastructure of TJ after Cd +A or Cd +SB203580 treatment .The positive products of zonula occludens-1 ( ZO-1 ) and claudin-11 were localized mainly in the base of the seminiferous tubule .After Cd treatment , the average absorbance (AA) of ZO-1 and Claudin-11 was decreased significantly compared with that of the control group (P<0.05).After Cd +A or Cd +SB203580 treatment,AA of ZO-1 and Claudin-11 were increased significantly compared with that of the Cd group(P<0.05),though lower than that of the control group .The result of Western blotting showed that phosphorylation-p38MAPK in Cd group was increased significantly compared with that of the control group (P<0.05).After Cd +A or Cd+SB203580 treatment, it was decreased significantly compared with that of the Cd group (P<0.05).Conclusion Cd decreases ZO-1 and Claudin-11 expression and damages ultrastructure of TJ in BTB , asⅣhas protective effect on it , and is related to inhibiting activation of p 38 MAPK pathway .

Objective:To evaluate the short-term prognostic value of model for end-stage liver disease (MELD) combined with high density lipoprotein-cholesterol (HDL-C) in patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF).Methods:From December 2015 to December 2018, 182 patients with HBV-ACLF who were treated in Henan Provincial People′s Hospital were included. Prognosis and clinical data including HDL-C, total bilirubin, international standardized ratio (INR), creatinine of patients within 24 hours after admission were collected and analyzed retrospectively.The values of MELD were calculated. The binary logistic regression analysis was used to analyze the independent risk factors affecting 90-day mortality in HBV-ACLF patients.The receiver operator characteristic curve (ROC) and MedCalc 15.2 software were used to assess the predictive value of MELD, HDL-C and MELD-HDL-C model for prognosis. Kaplan-Meier survival curve was performed to analyze the prognosis of patients in different groups.Results:Sixty patients were divided into the death group and 122 patients were divided into the survival group according to the prognosis during hospitalization and 90 days after discharge. The MELD score of patients in the survival group was 21(19, 24), which was significantly lower than that in the death group (29(25, 34)), and the HDL-C value of patients in the survival group was significantly higher than that in the death group (0.3 (0.1, 0.6) mmol/L vs 0.2(0.1, 0.5) mmol/L). The differences were both statistically significant ( Z=-6.290 and -4.087, respectively, both P<0.01). Multivariate logistic regression analysis showed that MELD score and HDL-C value were the independent risk factors for 90-day mortality in patients with HBV-ACLF(odds ratio ( OR)=1.432, 95% confidence interval ( CI)1.271-1.613; OR=0.584, 95% CI 0.487-0.700, respectively; both P<0.01). Areas under the ROC of MELD, HDL-C and MELD-HDL-C scoring models were 0.775, 0.782 and 0.878, respectively. MELD-HDL-C scoring model was superior to both MELD and HDL-C , and the differences were both statistically significant ( Z=3.944 and 3.104, respectively, both P<0.01). When the MELD-HDL-C Youden′s index was set at 0.72, the optimal threshold was 24.69. Patients with MELD-HDL-C score≥24.69 had lower survival rate than patients with MELD-HDL-C score<24.69, and the difference was statistically significant ( χ2=142.900, P<0.01). Conclusion:MELD, HDL-C and MELD-HDL-C scoring systems could predict the short-term prognosis in patients with HBV-ACLF, and the predictive value of MELD-HDL-C has the superiority.

In this paper, five different methods were carried out for DNA extraction directly from soil. The result shows that all five methods could generate DNA with more than 15 kb in size. They were subsequently used as templates for PCR amplification with success, using primers of the bacterial 16S rRNA gene and Shiva-1 gene encoding an antibacterial peptide. However, method 5 is more suitable for DNA extraction directly from a small amount of soil sample as it produced a good yield of DNA in high integrity with reliable reproducibility.

Objective To investigate the toxic effect of cadmium (Cd) on the ultrastructure ,expression of related protein and the signal molecule phosphrylated P38 mitogen-actived protein kinase(P-P38MAPK) of primary cultured rat sertoli cell(Sc) ,and the protective effect of astragaloside (A ) on it .Methods The primary cultured rat Sc were divided into the control group ,Cd (50 mol/L)group and Cd(50 mol/L) plus A(10 mg/L) group ,they were used for the electron microscope observation and the im-munohistochemistry detection of vimentin ,E-cadherin ,-catenin and P-P38MAPK .Results The Sc ultrastructural changes included that the swelled mitochondria ,abundant lipid droplets and dilated endoplasmic reticulum were found in the Cd group .Further ,apop-tosis occurred in some Sc .However these ultrastructure changes above mentioned were slighter in the Cd plus A group ;the immu-nohistochemistry showed that the positive products of vimentin ,E-cadherin and-catenin were obviously decreased in the Cd group (P<0 .05) ,and those in the Cd plus A group were higher compared with the Cd group(P<0 .05);the expression of P-P38MAPK in cytoplasm was increased in the Cd group ,and showed the trend to move from cytoplasm to nucleus ,meanwhile ,the positive prod-ucts expression in the Cd plus A group was lower than that in the Cd group (P<0 .05) .Conclusion Cadmium can cause the injury of the Sc ultrastructure ,damage of cytoskeletal protein and fibronectin ,and increase of P-P38MAPK level ;astragaloside can antago-nize the toxicity of cadmium on Sc ,the protective effect maybe related with the decrease of P-P38MAPK in Sc .

Objective To investigate the virological response in hepatitis C virus (HCV)genotype 1b relapsers after 48 weeks of peginterferon/ribavirin (peg-IFN/RBV)combination retreatment,and to explore the predictive value of interleukin (IL )-28B rs12978960 genetic polymorphismon virological response.Methods From 2012 to 2014,genotype 1b chronic hepatitis C (CHC)relapsers in He′nan Provincial People′s Hospital were retreated with combined peg-IFN/RBV for 48 weeks and followed up for 24 weeks off-treatment.Host IL-28B genetic polymorphism was detected.Predictive factors associated with virological response and sustained virological response (SVR)were analyzed.Independent-samples t test was conducted in continuous variables,whileχ2 test or Fisher exact probability test was conducted in counts data.Results A total of 61 patients finished 48 weeks of peg-IFN/RBV combination therapy and were further followed up for 24 weeks off-treatment.Mean age was (46.7 ±12.4)years.Thirty-seven patients (60.7%)were male and 49 were rs12978960 CC genotype.After 48 weeks of retreatment with peg-IFN/RBV and 24 weeks of off-treatment follow-up,40 patients (65 .6%)achieved SVR.Rapid virological response (RVR)and SVR of younger patients were both significantly higher than those of older patients (100.0% vs 67.4% and 85 .0% vs 47.6%,respectively;both P =0.006).IL-28B rs12978960 genotype was predictive to RVR and SVR.Patients with RVR and SVR had higher carriage rates of IL-28B rs12978960 CC genotype compared with those without RVR and SVR (both P <0.05 ).Patients with CC genotype had higher rates of RVR (34.1 % vs 0;χ2 = 10.625 ,P =0.006 ),end-of-treatment virological response (84.1 % vs 70.6%;χ2 =5 .563,P =0.039 )and SVR (77.3% vs 35 .3%;χ2 =9.572,P =0.007)than those with CT/TT genotype.However,there were no statistical differences of extended RVR (34.1 % vs 29.4%;χ2 =0.122,P =0.809)and early virological response (79.5 % vs 82.3%;χ2 =0.612,P =0.964).Conclusions Retreatment with antiviral therapy is necessary in CHC patients with genotype 1b. IL-28B rs12978960 genetic polymorphism is predictive to the SVR of retreatment,especially for patients without RVR,which will provide individualized treatment and optimize the treatment strategy.

Leptin, the product of obese gene, is a secreting protein that exerts multiple biological functions by binding to its receptor. Leptin regulates nutrient intake and metabolism, and is secreted from adipocytes, which occupy most of the bone marrow cavity and constitute the microenvironment. Leptin not only plays an important role in the control of the proliferation and differentiation of normal primitive hematopoietic cells, but it also stimulates the growth and viability of leukemic cells. Leukemic cells of some patients with acute myeloid leukemia (AML), acute lymphoblastic leukemia, and chronic myeloid leukemia also express the leptin receptor. Furthermore, leptin also stimulates leukemic cell growth in vivo by promoting angiogenesis. These findings suggest the possibility that leptin and its receptor play roles in the pathophysiology of leukemia, and blockage of leptin binding to its receptor might have potential therapeutic benefits in the treatment of certain leukemias. This review discusses the biological characteristics of leptin and its receptor, the relation of leptin and its receptor with normal hematopoiesis, the relation of leptin and its receptor with AML and so on.
Animals ; Hematopoietic System ; Humans ; Leptin ; physiology ; Leukemia, Myeloid, Acute ; Receptors, Leptin ; physiology

OBJECTIVETo construct a retroviral-mediated vector of FLT3 ligand (FL) and express it in human bone marrow stromal cells.

METHODSFL cDNA was inserted into the retroviral vector pLXIN by gene recombination technology. The recombinant plasmid pLFIN was transferred into retrovirus packaging cell line PA317 by lipofectamine, and the positive clones were selected by G418. The mRNA expression in human stromal cells and integration of genome DNA were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR) and genomic DNA-PCR. The expression of FL protein and its biological activities in the culture were investigated by ELISA and mouse bone marrow CFU-GM assay.

RESULTSThe recombinant plasmid pLFIN was successfully constructed. In genome of these transfected target cells, Neo gene and FL gene were integrated, FL mRNA was transcripted and FL protein was expressed at 4.35 ng/ml/24 h. The specific activities of FL in the culture indicated that human bone marrow stromal cells transfected with FL could significantly express FL in vitro.

CONCLUSIONThe retroviral-mediated FL gene was expressed in bone marrow stromal cells and the biological activities of FL were detectable in the supernatant of the transfected cells. These results provide a basis for studies on hematopoietic regulation by gene transfected bone marrow stromal cells.

3T3 Cells ; Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Line ; Colony-Forming Units Assay ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; Membrane Proteins ; genetics ; metabolism ; Mice ; RNA, Messenger ; genetics ; metabolism ; Retroviridae ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stromal Cells ; cytology ; metabolism ; Transfection

OBJECTIVETo explore the clinical features and gene mutation profiles of patients with chronic hepatitis B (CHB) and Gilbert's syndrome.

METHODSThirty-three patients with CHB and Gilbert's syndrome were enrolled in the study. Serum markers of liver function and histological features of disease-related liver injury were assessed by standard methods. Gene mutations were detected by PCR and direct DNA sequencing.Statistical analysis was carried out with the chi-square and t tests.

RESULTSSequencing of the Gilbert syndrome-associated gene, UGT 1A 1, revealed mutations in the upstream promoter phenobarbital-responsive element module (PBREM) (-3279 mutation, 23 cases), in the promoter TATA box (a TA insertion mutation, 21 cases), and in the coding region of exon 1 (a GGA-AGA Gly71Arg mutation, 18 cases); there was no statistical difference found for any of the three mutations among this patient population (x2 =1.640, P more than 0.05).

CONCLUSIONThe traditional methods of diagnosis for patients with CHB and Gilbert's syndrome remain a technical challenge in the clinic, and gene detection may represent a more favorable method for diagnosing this patient population.

Base Sequence ; Exons ; Gilbert Disease ; Glucuronosyltransferase ; Hepatitis B, Chronic ; Humans ; Mutagenesis, Insertional ; Mutation ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; TATA Box

OBJECTIVETo investigate the effects of adefovir dipivoxil (ADV) on blood phosphorus metabolism in patients with chronic hepatitis B (CHB).

METHODSPatients with hepatitis B surface antigen (HBsAg)-positive CHB were treated with ADV alone, ADV combined with interferon (IFN), or ADV combined with lamivudine (LAM). Changes in levels of calcium, phosphate, urea, and creatinine were assessed at treatment weeks 4, 12, 24, 48, 72 and 96. Statistical analysis was carried out with SPSS 16 software; influential factors were analyzed by ANOVA and non-conditional logistic regression analysis.

RESULTSDuring the course of treatments, 32 (42.6%) of the patients presented with low phosphorus. The highest incidence of low phosphorus was found to have occurred at treatment week 24 (25.0%, 27.5% and 36.4% respectively, with no statistical difference between three groups, x2=0.225, P>0.225). Patients with hypophosphatemia did not show a significant difference in serum phosphorus levels from the other patients (F=1.853, P=0.169). Logistic regression showed a correlation between low phosphorus and sex (x2=7.876, P<0.05), age (t=2.479, P<0.05), and serum creatinine (t =-2.256, P<0.05), but not with blood urea nitrogen or blood calcium (P>0.05).

CONCLUSIONADV antiviral treatment can decrease the blood phosphorous levels of CHB patients, particularly over extended time of treatment, and the occurrence of low phosphorus is more common than of mild phosphorus decrease.Male and elderly patients may be at greater risk of this complication. The incidence and severity of low phosphorus is not significantly different for the different ADV-based treatment regimens.

Adenine ; analogs & derivatives ; Aged ; Antiviral Agents ; Creatinine ; Drug Therapy, Combination ; Hepatitis B, Chronic ; Humans ; Interferons ; Lamivudine ; Male ; Organophosphonates ; Phosphorus

OBJECTIVETo isolate and culture human placenta derived adherent cells (hPDAC) and assay their hematopoietic growth factor expression.

METHODSBy enzyme digestion, hPDAC were isolated from human placenta tissue and cultured, and their biological characteristics were studied. The hematopoietic growth factor (HGF) mRNA expression of hPDAC was assayed by RT-PCR.

RESULTShPDAC was successfully isolated from human placenta tissue, which was further confirmed as mesenchymal stem cell-like cells. HGF including SCF, FL, G-CSF, GM-CSF, M-CSF and IL-6 were expressed in hPDAC.

CONCLUSIONhPDAC could be used as feeder layer for umbilical cord blood CD(34)(+) cells ex vivo expansion.

Cells, Cultured ; Female ; Gene Expression ; Granulocyte Colony-Stimulating Factor ; genetics ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Hematopoietic Cell Growth Factors ; genetics ; Humans ; Interleukin-6 ; genetics ; Macrophage Colony-Stimulating Factor ; genetics ; Membrane Proteins ; genetics ; Placenta ; cytology ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics