Objective:To investigate L-type calcium channels of human mesenchymal stem cells(hMSCs) cultured in vitro.Methods:hMSCs were isolated,cultured by Percoll gradient centrifugation,adherence to plastic flask and monoclonal culture.Immunophenotypes of hMSCs were detected by immunofluorescence and flow cytometry techniques.Whole cell patch-clamp technique was used to observe the change of L-type calcium channels of human mesenchymal stem cells(hMSCs).Results:High homogenous hMSCs had been isolated and cultured in vitro.hMSCs had unique immunophenotypes and they were positive for CD44,CD29,c-kit but negative for CD34,CD31,CD54.Calcium currents could be found in 40 percent of hMSCs,peak currents of calcium(I Ca-Peak ) were (102.67?19.06)pA from +20 mV to +30 mV.The currents could be inhibited by Cd~ 2+ (20 ?mol/L).Conclusion:Undifferentiated hMSCs express less L-type calcium channes,regulation of the chanels might contribute to directional differentiation.

Objective To investigate the effect of controlled low central venous pressure(CLCVP) on blood loss and prognosis in different types of hepatectomy .Methods Two hundred and fifty seven patients underwent standard hepatectomy ,half liver resec‐tion or irregular partial hepatectomy from January 2011 to December 2012 in the First Affiliated Hospital of Chongqing Medical U‐niversity were retrospectively studied .Patients treated with CLCVP during hepatectomy were attributed to the CLCVP group .CVP of these patients were lowed to below 5 cm H2 O by minimizing fluid infusion and one or both of the following maneuvers :posture adjustment ,nitroglycerin administration .Alpha agonists were used when necessary to maintain the mean arterial pressure MAP at ≥60 mm Hg .Other patients been maintained with normal level of CVP by adjusting fluid administration were included in normal CVP group (NCVP) .Blood loss and transfusion volume ,length of hospital stay of the two groups were compared ,and the effects of different surgery type on CLCVP blood protection were evaluated .Results In the patients underwent standard hepatectomy or half liver resection ,intraoperative blood loss and transfusion were not statistically different between the two groups .While in the pa‐tients underwent irregular partial hepatectomy ,the CLCVP group suffered less blood loss and transfusion(P<0 .05) .Percentage of the patients with less than 200 mL blood loss and no transfusion of concentrated red cell in CLCVP group was higher than that of in NCVP group(P<0 .05) .Differences between the two groups in postoperative hospital stay were with no significance in all the operation types(P>0 .05) .Conclusion The efficiency of CLCVP on blood protection during hepatectomy is influenced by the sur‐gery type ,the blood protection is found to be significant only in irregular partial hepatectomy .No relationship was found between CLCVP and postoperative hospital stay in all types of hepatectomy .

Objective To evaluate the clinical efficacy and safety of capecitabine continuous maintenance in treatment of re-currence of triple-negative breast cancer,which had responded to combined chemoradiotherapy.Methods The triple-negative breast cancer was defined as negative of ER,PR and HER2.A total of 46 patients of triple-negative breast cancer were divided into the treatment group (23 patients)and the control group (23 patients).The treatment group was given capecitabine continuous mainte-nance after 6 cycles of chemotherapy.The control group was given 6 cycles of combined chemotherapy only.The clinical efficacy and safety was evaluated between the two groups.Results The PR,PD,RR,and DCR in the treatment group were significantly higher than those in the control group(P <0.05).The acute toxic effects (except for hand-foot syndrome)were similar in two group(P >0.05).Conclusion Capecitabine continuous maintenance after combined chemoradiotherapy in treatment of recurrence of triple-neg-ative breast cancer is more effective,lower toxicity and tolerable.

OBJECTIVETo investigate the infection of HEV in Quzhou area of Zhejiang Province.

METHODSAll sera from blood donors in the central blood bank of Quzhou from April 2006 to April 2007 were used. Anti-HEV IgG and anti-HEV IgM were measured by EIA. RT-PCR was also performed to the samples with positive anti-HEV IgM. Genotype and sequence homology were analyzed after sequencing.

RESULTSThe positive ratio of anti-HEV IgG was 40.60%, in which the male infection ratio was higher than the female significantly (43.09% VS 36.09%; chi2=22.6; P < 0.01). The infection ratio was increased with age. The positive ratio of anti-HEV IgM was 0.43%. The positive ratio of anti-HEV IgG and the titers of antibody were higher in the inferior clinical infectors with positive anti-HEV IgM than the negative ones (P < 0.05). Two samples were positive in HEV PCR among 21 samples with positive anti-HEV IgM. The toxemia ratio was 0.4% of all the donors. And the genotype of the two samples with toxemia were both HEV-IV.

CONCLUSIONThe HEV infection was correlation with age and sex significantly and the infection occurred in the adults mainly in Quzhou area. HEV toxemia was not infrequency in the blood donors.

Adolescent ; Adult ; Age Factors ; Antibodies, Viral ; analysis ; immunology ; Blood Donors ; China ; epidemiology ; Female ; Genotype ; Hepatitis E ; complications ; epidemiology ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; Humans ; Immunoglobulin G ; analysis ; immunology ; Immunoglobulin M ; analysis ; immunology ; Male ; Middle Aged ; Sequence Analysis, DNA ; Sequence Homology ; Sex Factors ; Toxemia ; complications ; epidemiology ; virology

There are sex differences in some brain areas in mammalians. Parkinson's disease (PD), caused by the mesencephalic substantia nigra (SN) dopaminergic neuronal loss, displays sexual difference, i.e., the incidence is higher and the symptoms are more intense in males than that in females. However, it has not been known whether sexual dimorphisms exist in the SN. Sixty adult Sprague-Dawley rats were randomly divided into 5 groups: (1) Female intact group (F-INT group); (2) Male intact group (M-INT group); (3) Ovariectomized group (OVX group); (4) Castration group (CAST group); (5) Ovariectomized + estrogen-replaced group (OVX-E(2) group): The rats received sequentially physiological dose of estrogen for 3 d from the 7th day after ovariectomization. P50 auditory evoked potential (P50) was recorded for 14 d from electrodes inserted in the rat right SN in quiet and awake state. After recording, the brain tissues were dissected and the tyrosine hydroxylase (TH)-expressing neurons in the compact zone of the SN were counted using immunohistochemical method. The results showed that the number of TH-positive (TH(+)) cells in the SN of normal male animals was less than that in normal female rats (P<0.05), and the T/C ratio of P50 in normal males was significantly less than that in normal females (P<0.01), indicating that there exists sexual difference in function and structure in the SN. No differences in the T/C ratio of P50 or the number of TH(+) cells were found between M-INT and CAST groups. The T/C ratio of P50 and the number of TH(+) cells in the SN in OVX group were reduced significantly compared with those in F-INT group (P<0.01). There was no significant difference in the T/C ratio of P50 and the number of TH(+) cells in the SN between OVX- E(2) and F-INT groups 15-20 d after estrogen replacement, suggesting that estrogen can promote the survival and functional recovery of dopaminergic neurons in the SN. The results suggest that there exist sex-specific differences in the dopaminergic neurons in the SN structurally and functionally. The difference of estrogen level in cerebra between male and female animals may account for the sexual differences. Endogenous estrogen plays an important role in maintaining the integrity and modulating the functional activity of dopamine system in the SN.
Animals ; Dopaminergic Neurons ; cytology ; Estrogens ; pharmacology ; Evoked Potentials, Auditory ; Female ; Male ; Orchiectomy ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Sex Characteristics ; Substantia Nigra ; cytology

OBJECTIVETo measure the plasma levels of transforming growth factor beta1 (TGFbeta1), the protein expression of alpha-SMA in hepatic stellate cells and urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1), and study on the relationships between the plasma levels of TGFbeta1, the protein expression and the serum hyaluronic acid (HA) in patients with different grades of hepatic fibrosis.

METHODSThirty seven cases with hepatic fibrosis of different grades were classified according to HE and VG staining categories from 0 to 4, in which there were 8 cases in grade 1, 9 cases in grade 2, 7 cases in grade 3, 13 cases in grade 4. The plasma levels of TGFbeta1 and the serum levels of HA were detected by ELISA. The protein expressions of a-SMA, uPA and PAI-1 in fibrotic liver tissue were observed by immunohistochemistry.

RESULTSWith the progression of hepatic fibrosis, the plasma levels of TGFbeta1 and the protein expression of a-SMA, uPA and PAI-1 in fibrotic liver tissue were increased. In grade 3 and 4, the plasma levels of TGFbeta and the protein expression of a-SMA and PAI-1 in fibrotic liver tissue were significantly increased, but the protein expression of uPA in cirrhosis liver tissue did not increased.

CONCLUSIONTGFbeta1, a-SMA, uPA and PAI-1 play an important role in the progression of hepatic fibrosis. Inhibiting the early activation of latent TGFbeta1 or increasing uPA and inhibiting PAI-1 over express may contribute to matrix degradation and retard the progression of hepatic fibrosis.

Actins ; blood ; Adult ; Aged ; Female ; Hepatitis B, Chronic ; blood ; complications ; Humans ; Liver Cirrhosis ; blood ; etiology ; Male ; Middle Aged ; Plasminogen Activator Inhibitor 1 ; blood ; Transforming Growth Factor beta ; blood ; Transforming Growth Factor beta1 ; Urokinase-Type Plasminogen Activator ; blood

According to the successful experience of studying on crops, this paper introduced the conception of the core collection of medicinal plants, and analyzed the characteristic and the way to construct it. Studies on the core collection would fulfill the management and utilize the germplasm resources conveniently and provide a new idea and a method to study on the germplasm resources of medicinal plants. It is necessary to study on the core collection for the development of the germplasm resources of medicinal plants.
Chromatography, High Pressure Liquid ; Conservation of Natural Resources ; Culture Techniques ; Ecosystem ; Plants, Medicinal ; chemistry ; genetics ; growth & development ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo establish the allele specific primer polymerase chain reaction sequence-based typing (PCR-SBT) and investigate the polymorphism of exon 3 of human leukocyte antigen( HLA)-DRB1.

METHODSThe fragment containing exons 2 and 3 of HLA-DRB1 gene was amplified by group specific primers. The amplified products were digested by restriction enzymes and directly sequenced in both directions. The genotype was assigned by using Assign 3.5 SBT software.

RESULTSThe exon 3 sequences of HLA-DRB1*08:09 and HLA-DRB1*12:02:01 were identified for the first time. There were 27 polymorphic sites in exon 3 among the twenty-five HLA-DRB1 alleles, which was 9.56% of all nucleotides of exon 3. The method could discriminate the HLA-DRB1*14:01:01/14:54 ambiguous samples, and the HLA-DRB1*14:01:01 was identified in the Chinese population.

CONCLUSIONThe PCR-SBT method for exon 3 of HLA-DRB1 from the present study was reliable and there were polymorphisms in exon 3 of HLA-DRB1.

Alleles ; Base Sequence ; DNA Primers ; genetics ; Evolution, Molecular ; Exons ; genetics ; Genotype ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; genetics

Objective To explore the expression of glutathione hormone (GSH) and γ-glutamyl cysteine synthetase (γ-GCS) in premature newborn rats exposed to hyperoxia,and to study antioxidant role of GSH and γ-GCS in hyperoxia-induced lung injury.Methods One-day old preterm SD rats were divided randomly into 2 groups:hyperoxia group and air group.Newborn rats in hyperoxia group were continuously exposed to oxygen(oxygen ＞ 850 mL/L),and newborn rats in air group were exposed in room air.After 1,7,14 days of exposure,the preterm SD rats of 2 groups were killed,and whole lung of these rats were isolated.GSH and γ-GCS of pulmonary tissue homogenate were detected by double antibody sandwich enzyme linked immunosorbent assay (ELISA).Bicinchoninic acid (BCA) was used to detect GSH protein in lung tissue homogenate.Total lung RNA was extracted and γ-GCS mRNA was detected by reverse transcription polymerase chain reaction.Results 1.The results were detected by ELISA method and BCA method,compaired with air group,the expression of GSH protein in lung tissue induced by hyperoxia was significantly increased after 1,7 days of exposure(all P ＜ 0.05),but the expression of GSH became significantly weak after 14 days of exposure (P ＜0.05).2.The expression of γ-GCS mRNA and protein level were significantly increased in 1,7 days (all P ＜0.05),but the general tendency decreased after 14 days of exposure,the expression of γ-GCS mRNA became stronger than its expression after 14 days of air group,both were no significantly different(P ＞0.05).Conclusions The changes of GSH and γ-GCS in the lung of premature SD rats induced by oxidation outbreak participate in the development of hyperoxia-induced lung injury,the activity of γ-GCS may be increased by hyperoxia,and alleviate hyperoxia lung injury in premature rats through antioxidation of GSH.

OBJECTIVETo establish the eukaryotic cell expression system for the alpha-1,2 fucosyltransferase gene FUT1 293C to T and 658C to T mutations and explore the mechanism of FUT1 mutations resulting in the reduced expression of H antigen.

METHODSGenomic DNAs were extracted from individuals with para-Bombay phenotype and full coding region of FUT1 was amplified. The amplification fragments were ligated with pcDNA3.1 plasmid to construct the recombinant expression vectors. The recombinant plasmids were transfected into the COS-7 cells using lipofectamine transfection reagent and stable expression screening was performed. FUT1 mRNA expression was determined by real-time quantitative PCR. The activity of enzyme was measured by high performance liquid chromatography. Expressed protein was identified by SDS-PAGE and Western blotting.

RESULTSpcDNA3.1-FUT1 wildtype, pcDNA3.1-FUT1 293C to T and pcDNA3.1-FUT1 658C to T recombinant vectors were constructed, respectively. COS-7 cells with stable expression of FUT1 were obtained through recombinant plasmid transfection and screening with G418. The FUT1 mRNA level of transfected cells with 293C to T and 658C to T recombinant vectors reached 97.10% and 104.74% of the wildtype FUT1 transfected cell. A specific protein band with about 46 000 was confirmed in the transfected cell lysates by SDS-PAGE electrophoresis and Western blotting with 6×His Tag antibody. The wildtype FUT1 transfected cell lysates can catalyze the enzymatic reaction, while the enzyme activity of cell lysates from 293C to T and 658C to T were abolished.

CONCLUSIONThe results suggested that the 293C to T and 658C to T mutations of FUT1 gene did not affect the RNA and protein expression levels, but the enzyme activity of cells with FUT1 mutations was significantly decreased which resulted in the reduced expressin of H antigen.

Animals ; COS Cells ; Cercopithecus aethiops ; Fucosyltransferases ; genetics ; metabolism ; Gene Expression Regulation ; Genetic Vectors ; genetics ; Mutant Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics