OBJECTIVETo explore effective pretreated methods for hydrosalpinx before frozen embryo transfer (FET).

METHODSA randomized controlled study was performed on 229 FET cycles of hydrosalpinx patients. They were assigned to two groups by random digit table, Group A (94 cases), Group B (89 cases), and Group C (46 cases). Patients in Group A received transvaginal aspiration of hydrosalpinx combined with auricular point sticking. Those in Group B received transvaginal aspiration of hydrosalpinx group. Those in Group C received no transvaginal aspiration of hydrosalpinx. Pregnancy outcomes of FET, endometrial and subendometrial blood flow distribution on the embryo transfer day were compared among the three groups.

RESULTSThere was no statistical difference in the endometrial thickness on FET day, the number of transfer embryos, the number of transferred good quality embryos among the three groups (P > 0.05). The clinical pregnancy rate and the embryo implantation rate were significantly higher in Group A than in Group C (P < 0.05), and the clinical pregnancy rate was significantly higher in Group A than in Group B (P < 0.05). The early abortion rate and the transfer cycle cancel rate were significantly lower in Group A than in Group C (P < 0.05). Type A endometrial and subendometrial blood flow distribution was dominant in Group A, which was significantly higher in Group A than the rest two groups (P < 0.05). Type A distribution rate was also significantly higher in Group B than in Group C (P < 0.05).

CONCLUSIONTransvaginal aspiration of hydrosalpinx combined with auricular point sticking before FET could improve the endometrial receptivity and improve outcomes of IVF.

Embryo Implantation ; Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; methods ; Humans ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate

Objective To observe the metabolism and distribution of arsenic in liver and brain of offspring rata by exposure to arsenic of pregnant rats or lactation dams and weaned pups,and explore if arsenic could penetrate the placental barrier,lactation barrier and blood brain barrier. Methods The Wistar female rots were randomly divided into four groups according to body weights,12 in each group,and were fed with drinking water that contained arsenic(NaAsO_2) 0,10,50,100 mg/L beginning from the gestafional day 6 until pups 42 days old. Pups were separately sacrificed on postnatal day(PND) 0,15,28,42. Arsenic in liver and brain of offspring rots and in breast milk was examined by atomic absorption speetrophotometer with an arsenic speeiation pretreatment system. Results Concentration of iAs,MMA,DMA of brain in 50,100 mg/L groups were higher than that of 0 mg/L group[0,0,0,(7.3±6.6),0,(44.2±27.4)ng/g]on PND 0,42[iAs: (120.0±46.0),(195.5±125.3),(216.5±278.4),(176.6±151.8) ng/g; M MA: (47.2±18.1),(199.6±389.1),(47.4±55.2),(82.7±79.2) ng/g; DMA: (984.3±377.4),(2222.1±1433.2),(998.1±368.3),(1781.3±715.7)ng/g,all P < 0.05]. Concentration of DMA of brain in 50,100 mg/L groups were higher than that of 0 mg/L group[(13.9±18.1),(50.6±98.3)ng/g]on PND 15,28 [(270.3±73.1),(323.9±72.7),(758.7±245.9),(1020.6±383.6) ng/g,all P < 0.05]. Concentration of iAs,DMA of liver in 10,50,100 mg/L groups were higher than that of 0 mg/L group [(1.4±3.5),(49.7± 47.1),0,(100.4±30.2)ng/g]on PND 28,42 [iAs: (37.5±28.1),(268.8±246.4),(307.2±339.9),(15.4±9.4),(479.1±161.1),(408.4±51.9)ng/g;DMA: (594.5±148.8),(3181.9±519.0),(4834.2±2568.4),(1061.8± 85.2),(3697.1±553.7),(4120.0±732.8) ng/g,all P < 0.05]. Concentration of DMA of liver in 10,50,100 mg/L groups were higher than that of 0 mg/L group[(13.2±20.5)ng/g]on PND 15[(182.0±60,2),(637.6±90.0),(1458.7±196.3)ng/g,all P < 0.05]. Concentration of arsenicals of liver and brain showed a dose-dependent increase. The concentrations of DMA of breast milk in 50,100 mg/L groups were also higher than that of 0 mg/L group[(9.8±13.4),0 ng/g]on PND 0,15 [(182.3±85.9),(372.2±203.9),(124.2±33.1),(244.4±196.5)ng/g,all P < 0.05]. In the analysis of the change of arsenic on different postnatal day,we found the concentration of iAs,MMA,DMA,TMA in liver and brain of pups all decreased on postnatal day 15,and was lower than that on PND 0,28 and 42. Conclusions The distribution of arsenic and methyl-metabolism in liver and brain of pups is related with arsenic exposure dose. Arsenic can penetrate the placenta and blood brain barrier easily and lactation can hinder arsenic intake in some extent.

Objective To observe the expression of STEAP4 gene(a novel obesity-related gene) during the period of human preadipocyte differentiation and to explore the relationship between the STEAP4 gene expression and adipocytes differentiation,adipogenesis.Methods Human preadipocytes were cultured and differentiated into the matured adipocytes in vitro.Adipocytes morphology and lipid accumulation were observed during this process.Total RNA was extracted from adipocytes at various time points (preadipocyte,Day 0,Day 4,Day 6,Day 8,Day 11,Day 14,and Day 17) and the level of STEAP4 mRNA expression was measured by fluorescent real-time quantitative reverse transcriptase-polyme-rase chain reaction(RT-PCR).Results The level of STEAP4 mRNA expression remained high in preadipocytes.In the presence of differentiation medium (Day 4),there was a transient upregulation in the expression of STEAP4 gene.After that,with the human preadipocytes being differentiated into matured adipocytes,the expression of STEAP4 mRNA was downregulated and reached the lowest level in fully differentiated adipocytes.There was a significant difference between any 2 detected phases in the level of STEAP4 mRNA expression (Pa

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This study was purposed to investigate the effect of 3'-azido-2', 3'-dideoxythymidine (AZT)on the proliferation and telomerase activity of human acute myeloid leukemia cell line KG-1a. The effect of proliferation was detected by MTT assay after the KG-1a cell were stimulated for 24, 48 and 72 h with different concentrations of AZT; telomerase activity was detected with TRAP-PCR-ELISA assay; RT-PCR was used to detect telomerase hTERT mRNA expression. The results showed that the proliferation of KG-1a cells was inhibited in a time and concentration dependent manner after exposure to AZT for 24, 48 and 72 h; the KG-1a cells decreased in S phase and increased in G(2)/M phase with the increasing of the concentration of AZT; telomerase activity and hTERT-mRNA expression in the experimental groups decreased after treated with AZT, which was positively correlated with concentration of AZT. It is concluded that AZT inhibits KG-1a cell proliferation and induces apoptosis, which maybe related with its decreasing the telomerase activity and hTERT mRNA expression.
Apoptosis ; drug effects ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Leukemia ; metabolism ; pathology ; Telomerase ; antagonists & inhibitors ; metabolism ; Zidovudine ; pharmacology

In order to investigate whether cytokine-induced killer (CIK) cells can induce apoptosis of bcr-abl(+) K562 cells, apoptosis of K562 cells and CEM cells induced by CIK cells, etoposide or camptothecin was detected with flow cytometry DNA assay. RT-PCR showed that K562 cells expressed the bcr-abl fusion gene, K 562 cells, K562 cells/etoposide or K562 cells/camptothecin groups showed no sub-G(1) peak. K562 cells/CIK cells group showed sub-G(1) peak (38.1%). CEM cells showed no sub-G(1) peak. CEM cells/camptothecin or CEM cells/etoposide groups showed sub-G(1) peak (23.5% or 32.3% respectively). CEM cells/CIK cells group showed sub-G(1) peak (45.4%). Etoposide or camptothecin did not induce apoptosis of K562 cells. CIK cells induce apoptosis of K562 cells. Bcr-abl fusion gene prevented apoptosis induced by etoposide or camptothecin, but did not prevent apoptosis induced by CIK cells. This property may support the observed adoptive immunologic effect of allogeneic bone marrow transplantation and donor lymphocyte transfusions of CML case relapsing after allogeneic bone marrow transplantation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; immunology ; Camptothecin ; pharmacology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Etoposide ; pharmacology ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; drug effects ; immunology ; metabolism ; Killer Cells, Lymphokine-Activated ; cytology ; immunology

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA

Objective To study the influence on the tumor after percutaneous intra-tumor injection of ~(32)P-GMS in liver cancer as well as its suitable dose.Methods 24 New Zealand rabbits were used to establish the animal model of VX-2 liver cancer,and divided into A,B,C and D groups with individually 37,74,111 and 148 MBq of ~(32)P-GMS being injected,respectively;and then pathological changes of tumor were observed by light and electron microscope respectively.Result The dose of ~(32)P-GMS was obviously correlated with the radioactivity damage of tumor cells.In the A and B groups,the tumor cells were not observed to disappear completely after injection of ~(32)P-GMS,but in C group,tumor cells were almost completely disappeared and surrounded by a lot of connective tissue.Although the tumor cells were found to disappear completely in D group,normal liver tissues were also involved.Conclusion Percutaneous intra-tumor injection of ~(32)P-GMS with suitable dose that may induce the tumor tissue to be maximally damaged and may also provide some significances to prevent the tumor metastasis.

OBJECTIVETo identify the type of the intermediate filament (IF) protein of Angiostrongylus cantonensis and analyze its tissue localization.

METHODSRecombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction, and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting. Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A. cantonensis by immunohistochemical analysis.

RESULTSThe antigen IF were correctly expressed and purified, and identified as a keratin located in the intestine wall and cytoplusma.

CONCLUSIONThe antigen IF is distributed in the intestine wall of A. cantonensis.

Angiostrongylus cantonensis ; cytology ; metabolism ; Animals ; Cell Nucleus ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Intermediate Filament Proteins ; classification ; genetics ; isolation & purification ; metabolism ; Protein Transport

BACKGROUNDA monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).

METHODSA phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.

RESULTSSix positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07 x 10(6) individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.

CONCLUSIONSThe results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.

Animals ; Antibodies, Helminth ; immunology ; Antibodies, Monoclonal ; immunology ; Antigens, Helminth ; blood ; Base Sequence ; Immunoglobulin Fragments ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Peptide Library ; Rabbits ; Schistosomiasis japonica ; diagnosis ; Sensitivity and Specificity ; Serologic Tests