Intrauterine growth retardation (IUGR) is one of the most commonly encountered developmental toxicity, which could lead to perinatal morbidity and mortality, be also extended from the fetus to adulthood, and seriously affect the quality of the population. Caffeine widely exists in a variety of daily beverages and some drugs. Its consumption is increasing year by year. Caffeine intake during pregnancy is one of the risk factors for IUGR. However, its mechanism of adverse outcome based on embryonic research is still unclear. In this paper, the possible mechanisms of caffeine-induced IUGR focusing on 3 important factors-the mother, placenta and fetus were explored. Caffeine's impact on the mother is the chronic activation of renin-angiotensin system; on the placenta, caffeine induces cell damage or the failure of the cell proliferation/apoptosis balance, leading to blockage of blood supply to the placenta; caffeine is also capable of directly affecting fetal development through interfering its neuroendocrine.

With the rapid development of turfgrass industry in our country,more and more attentions are paid to the genetic breeding of turfgrass. Biotechnology has great application potential in varietals improvement of tall fescue,which is commonly grown as cool-season turf grass in the temperate region. The establishment of tall fescue regeneration and genetic transformation systems was summarized. Meanwhile,the historical and present situations of tall fescue breeding are introduced. The part of tall fescue regeneration was expounded from different explants and approaches. While,the part of genetic transformations was expounded from different means. At last,problems and prospects of genetic transformation of tall fescue are discussed.

Objective To explore clinical efficacy and safety of intraoperative localization of occult insu -linoma by using step-by-step occlusion of the pancreas .Methods 22 cases of occult insulinoma patients admit-ted from Mar.2003 to May 2013 were given intraoperative localization by adopting the technology of step -by-step occlusion of the pancreas .Results All the 22 patients were successfully completed the segmental resection of pancreas.The average operation time was(120 ±50)min, and the average intraoperative blood loss was (100 ± 80)ml.No blood transfusion was needed.Blood glucose rose rapidly when insulinoma was located within the scope of occlusion, blood glucose remained unchanged when insulinoma was beyond the scope of occlusion ,and blood glucose dropped swiftly when insulinoma was on the point of occlusion .Two patients had postoperative short-term pancreatic fistula and they were cured by conservative treatment .No other complications occurred .The average hospitalization time was(12 ±5)d.The result was good during the followed up of 8 to 24 months.Con-clusion The technique of step-by-step occlusion of the pancreas for localization of occult insulinoma is effective supplement for conventional methods , worthy of promotion .

Objective To establish a mice model of cisplatin-induced ototoxicity and to investigate the effect of cisplatin on the expression of caspase-3 in mouse cochlea.Methods Totally 69 Kunming mice were randomly divided into control group,cisplatin 2.5mg/(kg?d) group,cisplatin 3.5mg/(kg?d) group and cisplatin 4.5mg/(kg?d)group.Mice were injected intraperitoneally for 5 days.Auditory brainstem response(ABR) was measured to observe the change of hearing.Envision method of immunohistochemistry was applied to detect the expression of caspase-3 in cochlea.Results The weight and hearing of mice in different dose cisplatin groups were declined significantly as compared with those in control group(P

Cell Line ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Interleukin-1 ; administration & dosage ; pharmacology ; P-Selectin ; metabolism

Objective To use ELISA detecting self‐developed hepatitis B five internal quality control products .Methods Hepa‐titis B five positive sera by the detection of ELISA were diluted through optimal ratio ,and were homemade indoor quality control materials .Results Self‐control materials and commodities were simultaneously detected by ELISA ,and the test results were com‐pared ,the two were no significant difference(P>0 .05);Self‐control materials continuously detected by ELISA ,its batch variation were less than 15% ,and stability was in line with the requirements .Conclusion Self‐developed hepatitis B five indoor quality con‐trol materials are made simply ,have good stability ,are satisfied control effect ,and have promotional value .

OBJECTIVETo investigate the factors influencing the pregnancy outcomes of artificial insemination with donor sperm (AID), improve the pregnancy rate, and evaluate the safety of the offspring.

METHODSWe retrospectively analyzed 7,761 cycles of AID for 5,109 infertile couples performed between July 1, 2005 and June 30, 2013 in the Center of Reproductive Medicine of Shenyang No 204 Hospital, the outcomes of pregnancy, and the incidence of birth defects.

RESULTSTotally, 2 252 clinical pregnancies were achieved by AID, in which the pregnancy rate per cycle was 29. 02% and the cumulative pregnancy rate was 44. 08%. The clinical pregnancy rate was remarkably higher in the females of ≤ 35 years than in those of > 35 years old (30.31% vs 20.18%, P < 0.01), in the women with < 5-year infertility than in those with > 5-year infertility (30.83% vs 28.16%, P < 0.01), and in the patients of the ovarian stimulation group than in those of the natural cycle group (33.22% vs 28.68%, P < 0.01) The clinical pregnancy rate was the highest in the first treatment cycle (29.87%), with statistically significant difference from the fourth cycle (23.61%) (P < 0.05), but not between the other cycles (P > 0.05). There were 28 cases of birth defects in the offspring (1.40%), including 6 cases (21.43%) involving the cardiovascular system, 4 (14.29%) involving the musculoskeletal system, 3 (10.71%) involving the urogenital system, 3 (10.71%) involving the central nervous system, 2 cases (7.14%) of cleft lip and palate, 2 (7.14%) involving the respiratory system, 2 (7.14%) involving the gastrointestinal digestive system, and other anomalies.

CONCLUSIONFemale age, infertility duration, and ovarian stimulation treatment are important factors influencing the clinical pregnancy rate of AID. Artificial insemination with cryopreserved donor sperm does not increase the incidence of birth defects, which is considered as a relatively safe technique of assisted reproduction.

Adult ; Cryopreservation ; Female ; Humans ; Infertility ; Insemination, Artificial, Heterologous ; methods ; Male ; Maternal Age ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Semen Preservation ; methods ; Spermatozoa ; Time Factors

OBJECTIVETo study the effect of herbal compound 861 (HB861) on expression and activity of nitric oxide synthase (NOS) in hepatic stellate cells (HSC), and to explore the feasibility of its application in preventing and treating the early portal hypertension.

METHODSHSC of HSC-T6 cell line (1 x 10(5)/ml) were cultured in dish with 95% O2 plus 5% CO2 under 37 degrees C for 24 hrs, then divided into 5 groups, 6 dishes in each group. Group A was the blank control group. To Group B-E, HB861 2 mg/ml, HB861 4 mg/ml, HB861 8 mg/ml, HB861 4 mg/ml + NW-Nitro-L-Arginine Methyl Ester (L-NAME)4 mg/ml were added separately, and continuously cultured for 24 hrs. NOS activity was measured using colorimetry, NO level was determined by nitrate reductase technique. The cells were fixed by 4% paraformaldehyde for 2 hrs for test HSC-T6 iNOS expression by immunocyto-chemical method.

RESULTSHB861 in 2 mg/ml, 4 mg/ml and 8 mg/ml could increase HSC-T6 NOS activity from 1.7 +/- 0.1 to 2.5 +/- 0.3, 3.5 +/- 0.4 and 3.7 +/- 0.9 respectively (P < 0.01), the NO levels in supernatant were increased in parallel from 56.1 +/- 4.8 to 90.7 +/- 4.6, 99.7 +/- 4.1 and 109.0 +/- 2.7 respectively (P < 0.01). L-NAME could not inhibit the effect of HB861 in increasing the synthesis and secretion of NO by activated HSC-T6. Immuno-cyto-chemical study showed that there was iNOS expression in cytoplasm, and which could be increased by HB861.

CONCLUSIONThe activated HSC-T6 showed positive iNOS expression, suggesting it could produce NO. HB861 could markedly increase HSC-T6 iNOS expression and NOS activity, enhance the NO synthesis and secretion, it also could inhibit the contractility of activated HSC by way of increase HSC to secrete NO, so as to lower the resistance in hepatic sinusoid, therefore would play important role in preventing and treating of early portal hypertension.

Animals ; Cells, Cultured ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; enzymology ; Hypertension, Portal ; prevention & control ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; biosynthesis ; metabolism ; Rats ; Rats, Sprague-Dawley

In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.

Objective:To express the B subunit of Shiga-like toxin type Ⅱ,and analyze its expression form and receptor-binding activity.Methods:The slt2b gene was obtained from EHEC O157∶H7 by PCR,and cloned to the expression vector pET22b(+).The genetically engineered bacteria pET22b(+)-stx2B/BL21 expressed the recombinant StxB after induced with IPTG.The renatured inclusion bodies were purified by ion exchange chromatography.The expression form of rStx2B was investigated by denaturing and native electrophoresis.The receptor-binding activity was confirmed by fluorescence detection and flow cytometer.Result:The constructed genetically engineered bacteria expressed the rStx2B at a high level.The purified protein was obtained after denaturation,renaturation and ion exchange chromatography.According to the denaturing and native electrophoresis,the rStx2B was expressed in a dimmer form,which consists of two monomers cross linked with disulfide bridge.The rStx2B showed good receptor-binding activity by Hela-binding assay.Conclusion:The genetically engineered bacteria were constructed successfully.The receptor-binding activity of rStx2B was independent of the pentamers.