Objective To investigate the expression of autoantibodies to the angiotensin Ⅱ type Ⅰreceptor (AT1-AA) and endothelin-1 (ET-1) in pregnant women's blood and explore their correlation with the pathogenesis of preeclampsia.Methods Ninety pregnant women who delivered from June 2011 to December 2011 in the First Affiliated Hospital of Zhengzhou University were chosen as the study objects.They were divided into mild preeclampsia group (n =30),severe preeclampsia group (n =30) and normal group (control group,n =30).The levels of AT1-AA and ET1 in maternal peripheral blood and umbilical cord blood were detected by ELISA,and the mRNA expression levels of AT1-AA and ET1 in placenta tissues were determined by reverse transcription (RT) PCR.Moreover,the correlation clinical indexes were detected and analysed.Results (1) The levels of AT1-AA and ET1 in maternal peripheral blood of preeclampsia [mild group:(114 ± 19) ng/L and (31 ± 9) ng/L,severe group:(145 ± 15) ng/L and (38 ± 10) ng/L] were both significantly higher than that of control group [(59 ± 5) ng/L,(17 ±4) ng/L].In addition,compared with mild group,the levels of AT1-AA and ET1 in severe group were significantly higher (P ＜0.05).(2) The levels of AT1-AA and ET1 in umbilical cord blood of preeclampsia [mild group:(105 ± 14) ng/L and (35 ±6) ng/L,severe group:(118 ± 14) ng/L and (40 ±5) ng/L] were significantly higher than that of control group [(61 ± 12) ng/L,(24 ± 5) ng/L].In addition,compared with mild group,the levels of AT1-AA and ET1 in severe group were significantly higher (P ＜0.05).(3) The mRNA expression levels of AT1-AA and ET1 in placenta tissues of mild group (0.313 ± 0.039,0.296 ±0.028) and severe group (0.568 ±0.052,0.577 ±0.046) were significantly higher than that in control group (0.198 ± 0.017,0.137 ± 0.012),and the levels in severe group were significantly higher than that in mild group (P ＜0.05).(4) There was an evident positive correlation between AT1-AA and ET1 levels of preeclampsia women's peripheral blood,umbilical cord blood and placenta (P ＜ 0.05).(5) The level of AT1-AA in umbilical cord blood of preeelampsia pregnant women was positively correlated with S/D value of umbilical artery (P ＜ 0.05),and negatively correlated with the weight of the birth and the placental (P ＜ 0.05).Conclusion The AT1-AA in the blood of pregnant women plays an important role in promoting the generation and development of preeclampsia by increasing the ET1 secretion.

Levels of lipids, lipoproteins and apolipoproteins in 59 patients with cerebral infarction were compared before and after therapy. Evening—Primrose—oil group (EPOG 32cases) was treated with EPO and routine drugs (low molecular dextran, piracetam, vitamine E), the routine—drug group (RDG 27cases) was treated only with above- mentioned routine drugs. After treating for one month the lab data demonstrated that the(HDL—c, HDL2—c and apo A—I levels in EPOG were significantly higher than)before treating (P0.05). All parameters above- mentioned in RDG had no significant change after treating.

Aim To study the protective effects of Paeoniflorin (PF) on dopaminergic neurons in brain slice of substantia nigra treated with MPP~+ and to investigate the transcription of alpha-synuclei (α-syn) mRNA.Methods The organatypic brain slice culture of substantia nigra prepared from neonatal SD rats was placed on Millicell-CM porous membranes and cultured to day-10.Then the cultrues of slice were treated with different concentrations (0.1,0.5,1.0 mmol·L~(-1)) of MPP~+ for 24 h.Some of the cultrues treated with 0.5 mmol·L~(-1) MPP~+ also received PF (1 or 10 μmol·L~(-1)).Slices cultured in normal medium were used as vehicle control.The tyrosine hydroxylase (TH) immunohistochemical staining with the cell counting was used to determine the dopaminergic neruons.The transcription of α-syn mRNA was examined by real-time quantitative RT-PCR.Results MPP~+(0.1,0.5,1.0 mmol·L~(-1)) exposure markedly decreased the number of TH~+ cells in a dose-dependent manner (P<0.05 or P<0.01) and sharply induced the transcription of α-syn mRNA (P<0.01) in slices treated with 0.5 mmol·L~(-1) MPP~+.The addition of PF (10 μmol·L~(-1)) to MPP~+-treated slices significantly increased dopaminergic neurons survival (P<0.01) and downregulated the transcription of α-syn mRNA significantly (P<0.05).Conclusion PF can effectively inhibit the injury of dopaminergic neurons induced by MPP~+ on brain slice of substantia nigra and downregulate the transcription of alpha-synuclein.

Objective To investigate the effect of dynamic regulation of negative pressure values in vacuum sealing drainage (VSD) on healing of soft tissue wounds.Methods From January 2013 to December 2015,115 patients were treated at our institute for open soft tissue defects of Gustilo-Anderson type ⅢaⅢc.They were 69 males and 46 females,aged from 17 to 76 years (average,44.7 years).They were divided into 4 groups to be subjected to different negative pressure values.Group A (n =29) was subjected to negative pressure values from-40 to-20 kPa,group B (n =29) to values from-80 to-60 kPa,group C (n =29) to values from-40 to-20 kPa for the first 72 hours followed by values from-80 to-60 kPa,and group D (n =28) to values from-60 to-80 kPa for the first 72 hours followed by values from-40 to -20 kPa.The 4 groups were compared in terms of drainage volume,wound shrinking,cleaning time,healing time and complications.Results The drainage volumes on the 7th day in groups B and D (4.89 ± 0.66 mL/cm2 and 4.33 ± 0.96 mL/cm2) were significantly higher than those in groups A and C (2.90 ± 0.67 mL/cm2 and 2.99 ±0.56 mL/cm2);the wound shrinking areas on the 14th day in groups B and D (14.16 ± 1.77 cm2 and 13.84 ± 1.65 cm2) were significantly larger than in groups A and C (12.57 ± 1.92 cm2 and 10.95 ± 1.37 cm2) (P ＜ 0.05).The cleaning time in group A was significantly longer than in the other 3 groups (P ＜ 0.05).The healing time in groups A and C was longer than in groups B and D (P ＜ 0.05).The incidences of pain in groups B and C were significantly higher than in groups A and D (P ＜ 0.05).The incidences of infection and blocking in group A were higher than in the other 3 groups (P ＜ 0.05).Conclusion Dynamic regulation of negative pressure values from-80 to-60 kPa for the first 72 hours followed by values from-40 to-20 kPa is an optimal choice for VSD management of soft tissue defects due to adequate drainage,obvious wound shrinking,quick cleaning and healing,and limited complications.

Objective:To construct nucleotide vaccine based on HSP65 gene of mycobacterium tuberculosis.Methods:The gene encoding heat shock protein 65 kilodalton was amplified from genome of mycobacterium tuberculosis H37Rv strain by polymerase chain reaction(PCR) and correctly inserted into corresponding sites of eukaryotic expression vector pcDNA3.1(-) after restriction endonuclease digestion.The experimental mice was vaccined by this recombinant plasmid DNA.Results:The gene fragment inserted into the vector pcDNA3.1(-) was the HSP65 gene of mycobacterium tuberculosis H37Rv strain,which was confirmed by partial nucleotide sequencing.There was specific antibody produce in experimental mice vaccined by the recombinant plasmid DNA and this could protect mice against infection of mycobacterium tuberculosis.Conclusion:The successful construction of recombinant eukaryotic expression vector based on HSP65 gene of mycobacterium tuberculosis,such recombinant plasmid DNA induced specific antibody production in experimental mice,lays the foundation for further researching in prevention and treatment of tuberculosis.

Studies on the variation of amylase, lipase and lrotease activity of Whitmania pigra in 0-6 months old using 3, 5-dinitro- salicylic acid colorimetry, right-nitrophenyl palmitate ester (ρ-NPP) colorimetry and folin-phenol method. The results showed that pro- tease activity remained low before 1.5 months old and with the highest activity in 2 months old, but after showing a small peak in 4 months, alkaline protease rapid declined. Amylase was low at born, then gradually increased the activity of the highest in 2.5 months old. Lipase with a strong vitality at birth, then 1 month with minimum and 2 months peaked, but appeared a small peak in 4 months old. In summary, only lipase exhibits strong activity at birth, lipase with the strongest activity in the digestive tract during develop- ment. Protease, lipase and amylase with the strongest activity at 2-3 months old, but were decreased after 4 months old.
Age Factors ; Amylases ; metabolism ; Animals ; Leeches ; enzymology ; Lipase ; metabolism ; Peptide Hydrolases ; metabolism

Objective To investigate the therapeutic effect of uteroplasty aided with temporary balloon occlusion of abdominal aorta in treating pernicious placenta previa combined with placenta percreta.Methods Sixty-two third-trimester gravidas who were diagnosed as pernicious placenta previa combined with placenta percreta by prenatal ultrasound and magnetic resonance imaging (MRI) in the First Affiliated Hospital of Zhengzhou University from January 1,2013 to May 31,2016 were enrolled in this study.All of them received cesarean section and then underwent uteroplasty following temporary balloon occlusion of abdominal aorta.Efficacy of that treatment was evaluated from the perspectives of blood loss,operation time,complications and postoperative recovery.All data were analyzed by descriptive analysis.Results All of the 62 cases were diagnosed with placenta percreta during operation including 10 cases (16.1％) with perimetrium invasion,46 cases (74.2％) with invasion to the muscular layer of bladder and six cases (9.7％) with bladder invasion.The average blood loss of all cases during and within 24 hours after operation was (1 377.3±605.2) ml and (140.6±66.3) ml,respectively.The average operation time was (72.3 ±24.5) min and the average length of postoperative hospital stay was (5.8± 1.6) d.The six cases of placenta percreta with bladder invasion received bladder repair.Sixty-one cases had their uterus preserved and the other one case had a sub-total hysterectomy due to amniotic fluid embolism.One woman developed phlebothrombosis in her lower limbs after operation.No intestinal or ureteral injury,puerperal infection,uterine ischemia necrosis or death was reported.In addition to three cases lost to follow-up,the other 59 patients were followed up to May 31,2017.Results of physical examination indicated that the 59 cases had normal uterine involution after operation.Menses returned in 58 of the 59 cases without any difference from before,and did not return in one case due to breastfeeding.Conclusion Uteroplasty aided with temporary balloon occlusion of abdominal aorta is a safe and effective uterus-preserving surgery for patients with pernicious placenta previa combined with placenta percreta.

Objective To study the operative technique and effect of temporary balloon occlusion of the abdominal aorta for preventing intraoperative bleeding during cesarean for patients with pernicious placenta previa and placenta accreta. Methods Retrospective analysis was conducted on the intraoperative situation of forty-one cases and information of follow-up twenty-nine cases, which were pernicious placenta previa and placenta accreta and delivered in the First Affiliated Hospital of Zhengzhou University from May 1, 2013 to June 30, 2014. Diagnosis was confirmed by line of color Doppler ultrasound and MRI for all patients before operations. An interventional physician performed right femoral artery puncture and preset the abdominal aortic balloon catheter in the digital subtraction angiography operation room before cesarean. At the same time of fetal delivery, 10 ml normal saline was injected into the balloon immediately, which results in filling of the balloon and blocking of the aorta. According to the area and depth of placenta implantation and implantation or penetration of the posterior bladder wall, placenta separation, partial resection of the uterine wall and partial bladder resection and repair were performed correspondingly. Meanwhile, saline in the balloon was pumped out gradually until empty. Condition of placenta implantation, blood loss and blood transfusion volume during the operation, intraoperative and postoperative complications, the duration and dose of fetal radiation exposure, and Apgar score of neonates were analyzed. Results Among the 41 cases, penetrative placenta and implanted placenta were observed in five cases and 36 cases, respectively. The latter 36 cases including 28 cases of bladder posterior wall accreta and eight cases of bladder posterior wall penetration. For all cases, the average operation time was (68.5±15.3) min, the mean blood loss in the operation was (1 058±960) ml, among which eight received blood transfusion with an average of (600±400) ml, and the mean hospital stay was (8.2±2.3) d. Uteruses were reserved in all cases. The mean duration and dose of fetal radiation exposure was (8.1±3.6) s and (5.2±2.9) mGy, and the Apgar score of neonates was 8.7±0.5 at 1 min and 9.5±0.3 at 5 min, respectively. The patients were followed up until October 31, 2014. Among them, six were lost, six were still in puerperium, 18 were breast-feeding, and the menses of 11 had returned. Conclusion Preset abdominal aortic balloon catheter in pernicious placenta previa and placenta accrete patients might effectively reduce the blood loss during cesarean section as well as the risk of hysterectomy through temporary occlusion of the abdominal aorta.

This study aims to construct a eukaryotic expression system for envelope gene of Jaagsiekte sheep retrovirus, observes its localization in 293T cells, and investigates the potential in inducing malignant transformation of NIH3T3 cells. By RT-PCR, the full-length cDNA of envelope gene of Jaagsiekte sheep retrovirus (exJSRV-env) was amplified from the extract of naturally infected sheep lung. The clone of target gene was sub-cloned into eukaryotic expression system pEGFP-C1, and validated by PCR, restriction endonuclease, and sequencing. Bioinformatic analysis concerning biological function and cellular localiza tion of exJSRV-env was also performed. The recombinant clone of exJSRV-env was transfected into 293T cells and NIH3T3 cells by Lipofectamine LTX. The expression and celluar localization in 293T cells were validated by confocal microscopy. Soft agar colony formation assay was employed to test the anchorage-independent growth of NIH3T3. DNA sequencing and restriction enzyme digestion with Kpn I and Hind III indicated the correct construction of the recombinant plasmid, which was named pEGFP-C1/exJSRV-env. Amino acid sequence alignment of exJSRV-env with reference sequences found 85%-100% homogeneity. A YRNM motif was discovered at the cytoplasmic tail of envelope gene, which is exclusively found in exogenous viruses. Phylogenetic tree analysis showed that our clone of exJSRV-env clustered closely with pathogenic exogenous Jaagsiekte sheep retroviruses. Fluorescence microscopy indicated typical membrane localization of exJSRV-env protein. NIH3T3 cells transfected with exJSRV-env lost contact inhibition, and acquired colony forming ability in soft agar. This study indicated that envelope protein of Jaagsiekte sheep retrovirus can induce malignant transformation of mouse fibroblast cell NIH3T3. Discoveries of this study provide a basis for further structural and functional research on Jaagsiekte sheep retrovirus envelope protein.
Amino Acid Sequence ; Animals ; Betaretrovirus ; chemistry ; classification ; genetics ; physiology ; Cell Transformation, Viral ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Molecular Sequence Data ; NIH 3T3 Cells ; Phylogeny ; Retroviridae Infections ; veterinary ; virology ; Sequence Alignment ; Sheep ; Sheep Diseases ; virology ; Transformation, Genetic ; Tumor Virus Infections ; veterinary ; virology ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism

OBJECTIVETo study the oxygen consumption rate and suffocation point of Whitmania pigra.

METHODUsing the method of respiratory chamber, the oxygen consumption rate, oxygen consumption volume and suffocation point were obtained.

RESULTThe oxygen consumption rate(OCR) ranged from 0.049 to 0.094 mg x g(-1) x h(-1), the oxygen consumption volume(OCV) from 0.44 to 0.67 mg x p(-1) x h(-1), and suffocation point(SP) from 0.9 to 1.51 mg x L(-1), when the average weight of W. pigra was 10 g and the water temperature varied from 15 to 35 degrees C. OCR ranged from 0.044 to 0.058 mg x g(-1) x h(-1), OCV from 0.19 to 0.77 mg x p(-1) x h(-1) and SP from 1.4 to 1.57 mg x l(-1) when water temperature was 20 degrees C.

CONCLUSIONOCR rose when the water temperature and weight increased, but SP was justly opposite. The weight was negatively correlated with OCR and positively with OCV. OCR had little difference from day to night.

Animals ; Leeches ; metabolism ; physiology ; Oxygen ; metabolism ; Oxygen Consumption ; physiology ; Temperature ; Water