ObjectiveTo investigate the effect of tertamethylpyrazine on perioperative humoral immune function in patients required for autologous blood transfusion.MethodsSixty ASA Ⅰ or Ⅱ patients aged 20-60 yr weighing 50-75 kg undergoing elective spinal surgery were randomly divided into 2 groups ( n =30 each):test group and control group.In test group,tertamethylpyrazine 2 mg/kg was infused intravenously over 5 min at 30 min before the autologous blood was collected.Tertamethylpyrazine was added to the heparinized saline and washing saline at the same time (25 mg per 500 ml) until the final concentration reached 0.005 ％.Tertamethylpyrazine was not given in control group.Venous blood samples were taken before anesthesia induction (T0) and at 1 h after operation (T1),and on day 1 and 5 after operation (T2.3) for measurement of serum IgG and IgM concentrations by ELISA.The operation time,intraoperative blood loss and amount of salvaged blood reinfused were recorded.ResultsThere was no significant difference in the operation time,intraoperative blood loss and amount of salvaged blood reinfused between the two groups (P ＞ 0.05).Compared with the baseline value at T0,serum IgG and IgM concentrations were significantly decreased at T1-3 in control group and the serum IgG concentration was significantly decreased at T1.2 in test group ( P ＜ 0.05 or 0.01 ).The serum IgG concentration at T2.3 and serum IgM concentration at T1-3 were significantly higher in test group than in control group ( P ＜ 0.05 or 0.01 ).ConclusionTertamethylpyrazine can reduce humoral immunosuppression to some extent and improve the balance of humoral immunity in patients required for autologous blood transfusion.

Objective To investigate the effect of ulinastatin on the postoperative immune function of effective T-lymphocytes (Teff cells) and regulatory T-lymphocytes (Treg cells) in the peripheral blood in patients undergoing laparoscopic operation.Methods Forty ASA physical status Ⅰ or Ⅱ patients,aged 20-40 yr,weighing 45-65 kg,scheduled for elective laparoscopic operation for infertility,were equally and randomly divided into 2 groups using a random number table:control group (group C) and ulinastatin group (group U).At 15 min before induction of anesthesia,ulinastatin 200,000 U (in 100 ml of normal saline) was infused intravenously in group U,while the equal volume of normal saline was infused instead in group C.Anesthesia was induced with iv injection of fentanyl,propofol,and cisatracurium and maintained with iv infusion of propofol and sevoflurane.Before surgery (T0),at the end of surgery (T1) and on 1st and 3rd days after surgery (T2.3),venous blood samples were collected from the peripheral blood for determination of the expression of Teff cells (CD4 + CD25-) and Treg cells (CD4 + CD25+) and serum interferon-γ (IFN-γ) and transforming growth factor-β (TGF-β) concentrations (by ILISA).Teff/Treg and IFN-γ/TGF-β ratios were calculated.Results Compared with the baseline value at T0,the expression of Treg cells was significantly up-regulated,and Teff/Treg ratio was decreased at T2 and T3 in group C,and the expression of Teff cells was up-regulated at T3 (P ＜ 0.05 or 0.01),while no significant difference was found in the expression of Treg cells and Teff/Treg ratio at T3 in group U (P ＞ 0.05).Compared with group C,the expression of Teff cells was significantly up-regulated at T3,the expression of Treg cells was down-regulated and Teff/Treg ratio was increased at T2 and T3,the concentration of serum TGF-β was decreased and IFN-γ/TGF-β ratio was increased at T2 and T3 in group U (P ＜ 0.05 or 0.01).No significant changes were found in the serum IFN-γ and TGF-β concentrations and IFN-γ/TGF-β ratio at each time point in the two groups (P ＞ 0.05).Conclusion Ulinastatin can enhance the immune function of Teff cells in the peripheral blood and decrease the immune function of Treg cells,thus maintaining the balance of the postoperative cellular immune function in patients undergoing laparoscopic operation.

Objective To research the effect of ulinastatin on T-cytoimmunity in patients with infertility undergoing laparoscopic surgery. Methods Forty patients scheduled for receiving laparoscopic surgery were equally randomized into two groups, ulinastatin group (Group U) and control group (Group C). Ulinastatin was given to patients in the Group U at a dose of 20 × 104 U before anesthetic. No ulinastatin was given to patients in the Group C. Patients′venous blood samples for T-lymphocyte subset (CD3+,CD3+CD4+,CD3+CD8+) and CD3+CD4+/ CD3+CD8+ ratio calculation were collected before the surgery (T0) and at 0 h (T1),1st day (T2),3rd day (T3) after the surgery. Results CD3+ had less difference at T1~3 compared with T0 in the Group C but raised obviously at T2~3 in the Group U. CD3+CD4+ were only raised at T3 compared with T0 but raised obviously at T2~3 in the Group U. CD3+CD8+ were raised obviously at T2~3 compared with T0 in the Group C but had less difference in the Group U. CD3+CD4+/CD3+CD8+ had less difference at T1~3 compared with T0 in the Group C but raised obviously at T3 in the Group U. Conclusion The application of ulinastatin in laparoscopic surgery could significantly produce protective effect on T-cytoimmunity.

Objective To compare the clinical effects of dynamic hip screw(DHS),Gamma interlocking intramedullary nail and proximal femoral nail antirotation (PFNA) in the treatment of intertrochanteric femoral fractures in elderly patients.Methods From January 2008 to June 2015,103 elderly patients with intertrochanteric femoral fracture were treated with DHS (DHS group,33 cases),Gamma nails (Gamma group,30 cases),or PFNA (PFNA group,40 cases).By the AO classification,there were 44 cases of type 31-Al,30 cases of type 31-A2 and 29 cases of type 31-A3.The 3 groups were compared in terms of incision length,operation time,intraoperative blood loss,fracture healing time,postoperative weight-bearing time,Harris scoring,and incidence of postoperative complications.Results PFNA group incurred significantly shorter incision length (5.4 ±0.5 cm) and operation time (70.8 ± 16.2 min) than DHS group (12.6 ±2.7 cm and 102.6±17.4min) and Gamma group (7.5±0.8 cmand93.0±35.9 min) (P ＜0.05).The intraoperative blood loss in PFNA group (163.2 ± 60.6 mL) was significantly less than in DHS group (280.5 ±89.8 mL) and in Gamma group (204.9 ±62.2 mL),and that in Gamma group was also significantly less than in DHS group (P ＜ O.05).PFNA group had significantly shorter weight-beating time (11.0 ± 0.8 weeks),fracture healing time(13.6 ± 1.5 weeks) and significantly higher Harris good to excellent rate (92.5％) than DHSgroup (13.3±1.0weeks,15.8 ± 1.2 weeks and 84.8％) and Gamma group (12.5±1.3 weeks,14.2 ± 1.0 weeks and 86.7％) (P ＜ 0.05).The incidence of postoperative complications in DHS group (21.2％)was significantly higher than in Gamma group(10.0％) and in PFNA group (7.5％) (P ＜ 0.05).Conclusions DHS,Gamma nail and PFNA are effective means for the treatment of intertrochanteric femoral fractures in the elderly.Intramedullary fixation,especially by PFNA,shows superiority in the clinical outcomes.

Objective To explore prokaryotic expression,purification of hemolysin coregulatory protein(Hcp)of Vibrio cholerae,and preparation of its polyclonal antibodies.Methods PCR was used to amplify Vibrio cholerae Hcp gene and clone it into pET28a vector to construct recombinant expression vector.The recombinant vector pET28a-hcp was transformed into E.coil BL21(DE3)for expression condition optimization and expression form identification.The soluble Hcp protein was purified by Ni-NTA column.The purified Hcp protein was used to immunize BALB/c mice to prepare polyclonal antibodies.The antibody titer was detected by indirect enzyme-linked immunosorbent assay(ELISA)to evaluate its immunogenicity.Western blot was used to analyze the specific recognition of antibodies to Hcp protein in Vibrio cholerae.Results The enzyme fragment digested by recombinant vector pET28a-hcp was consistent with the expected,the sequencing results were consistent with the Hcp gene sequence in the GenBank database,and the pET28a-hcp recombinant plasmid was successfully constructed.The recombinant plasmid was induced by isopropyl-β-D-thiogalactopyranoside(IPTG)to express the target protein with a relative molecular weight of 28 kD.The pure Hcp protein was obtained after purification by Ni-NTA column,and then Hcp polyclonal antibody(anti-Hcp)with a titer of 1∶512 000 could be obtained from immunized mice.Western blot results showed that anti-Hcp had specificity in recognizing Hcp protein in Vibrio cholerae.Conclusion The soluble expression of Hcp protein is successfully obtained,and high-titer polyclonal antibodies against Hcp are obtained after immunization of mice,which may lay a foundation for subsequent studies on the role of Hcp protein in the pathogenesis of T6SS in non-O1/non-O139 V.cholerae.

Objective To explore the efficacy and safety of high-dose second-generation fat emulsion usage on the very low birth weight premature infants.Methods A total of 88 premature infants with very low birth weight (VLBW) in Neonatal Intensive Care Unit (NICU)of Fujian Provincial Maternity and Children Hospital,Affiliated Hospital of Fujian Medical University from December 2013 to December 2014 were randomly divided into experimental group and control group,with 44 cases in each group according to the table of random number.The experimental group received intravenous nutrition with 200 g/L second-generation fat emulsion within 24 hours after birth,the initial dose was 2.0 g/(kg · d) with an increase of (0.5-1.0) g/(kg · d) daily,the maximum dose was 3.5 g/(kg · d);the control group received intravenous nutrition with 200 g/L second-generation fat emulsion 24 hours later after birth,the initial dose was 0.5 g/(kg · d) with an increase of 0.5 g/(kg · d) daily,the maximum dose was 3.5 g/(kg · d).The other intravenous nutrition methods were same.The general conditions at birth,blood biochemical parameters,growth parameters and complications were compared between the 2 groups.Results The mean value of intravenous nutrition duration,length of stay,the glucose infusion rates of postnatal days 6 and 7,the serum triglyceride levels of postnatal days 7,chest circumference of the fourth weeks,the incidence of the low triiodothyronine(T3) syndrome and parenteral nutrition associated cholestasis(PNAC) were (22.27 ± 7.17) d,(37.75 ± 12.28) d,(8.10 ± 0.92) mg/(kg · min),(8.49 ± 1.06) mg/(kg · min),0.18(0.03-0.59) mmol/L and (27.21 ± 1.62) cm in the experimental group respectively,but (27.36 ± 11.37) d,(44.36 ± 16.45) d,(7.98 ±0.79) mg/(kg · min),(8.22 ±0.76) mg/(kg · min),0.28 (0.07-0.99) mmol/L and (26.56 ± 0.96) cm in the control group,respectively,and the differences were statistically significant between the 2 groups (t =2.512,5.403,4.314,9.705,696.500,6.668,all P ＜ 0.05).The incidence of the T3 syndrome and parenteral nutrition associated cholestasis (PNAC) in the experimental group was 25.0％ (11/44 cases) and 0(0/44 cases),respectively,which were significantly lower than those in the control group[81.8％ (36/44 cases)and 9.1％ (4/44 cases)],and the differences were statistically significant between the 2 groups (x2 =28.542,5.736,all P ＜ 0.05).The 2 groups had no significant difference in the incidence rates of other complications such as necrotizing enterocolitis,infection,retinopathy of prematurity,bronchopulmonary dysplasia,and the duration of oxygen therapy and mechanical ventilation(all P ＞ 0.05).Conclusions The high-dose second-generation fat emulsion usage [the initial dose 2.0 g/(kg · d)] in VLBW infants is safe and well tolerated.Advisable parenteral nutrition support strategy can promote growth of VLBW infants,shorten the intravenous nutrition duration and length of stay,reduce the incidence of the low T3 syndrome and PNAC,which has no influence on the incidence rates of other complications.

[Objective]To investigate the protective effect of Zhuangtongyin on the Middle Cerebral Artery Occlusion(MCAO)model by modulating ferroptosis through the Nrf2-SCL7A11/xCT-Gpx4 pathway and its underlying mechanism.[Methods]C57BL/6J mice were randomly divided into Sham operation group(Sham),model group(MCAO),low-dose Zhuangtongyin group(ZTY-L),high-dose Zhuangtongyin group(ZTY-H),with 5 mice in each group.The MCAO group was modelled by silica gel embolization,the middle cerebral artery of mice was embolized for 1h,then the silica gel was pulled out and reperfusion was performed after 72 h;and the other operations in the Sham group were the same as those in the MCAO group except that the thread plug was not inserted.The neural function of mice was evaluated by Zea-Longa method.TTC staining was used to evaluate the volume of cerebral infarction.The level of brain injury was evaluated by HE staining and Nissl staining.Prussian blue staining and the expression of iron transport-related carrier receptors TfR1 and DMT1 on mRNA level was detected by qPCR to evaluate the iron ion deposition level in mice brain.The expression of lipid peroxidation-related gene ACSL4 on mRNA level was detected by qPCR,and the content of 4-HNE was detected by ELISA kit to evaluate the lipid peroxidation level of mice brain.The expressions of ferroptosis marker PTGS2 mRNA level was detected by qPCR.The expressions of Nrf2,SCL7A11/xCT,Gpx4 in mice brain tissue were detected by Western-blot and immunofluorescence.[Results]Zhuangtongyin improved the nerve function of mice after MCAO(P＜0.05)and the cerebral infarction volume of mice(P＜0.05)and alleviate the pathological injury of cerebral cortex cells after MCAO operation.Zhuangtongyin attenuated the accumulation of trivalent iron ions in the brain tissue of mice following MCAO.Additionally,Zhuangtongyin downregulated the expression of TfR1 and DMT1 mRNA(P＜0.001),a transporter associated with cellular iron ion uptake,in the brains of post-MCAO mice.Furthermore,Zhuangtongyin reduced levels of lipid peroxidation product 4-HNE(P＜0.001)and suppressed ACSL4 mRNA expression in brain tissue post-MCAO(P＜0.001).Besides,Zhuangtongyin downregulated the expression of PTGS2 mRNA(P＜0.001),in the brains of post-MCAO mice.Zhuangtongyin increased the expression of nrf2 into the nucleus(P＜0.001),and increased the expression of xCT and Gpx4 in neurons after MCAO(P＜0.001).[Conclusion]Zhuangtongyin can enhance the nerve function and reduce cerebral infarction volume in MCAO/R mice,alleviate the pathological damage of cerebral cortex cells,and modulate the expression of key signaling molecules in the Nrf2-SCL7A11/xCT-Gpx4 pathway.Therefore,it is suggested that the mechanism by which Zhuangtongyin improves MCAO/R injury in mice may involve regulating ferroptosis through the Nrf2-SCL7A11/xCT-GPX4 pathway.

In this study, the mobilomes of nine teleost species were annotated by bioinformatics methods. Both of the mobilome size and constitute displayed a significant difference in 9 species of teleost fishes. The species of mobilome content ranking from high to low were zebrafish, medaka, tilapia, coelacanth, platyfish, cod, stickleback, tetradon and fugu. Mobilome content and genome size were positively correlated. The DNA transposons displayed higher diversity and larger variation in teleost (0.50% to 38.37%), was a major determinant of differences in teleost mobilomes, and hAT and Tc/Mariner superfamily were the major DNA transposons in teleost. RNA transposons also exhibited high diversity in teleost, LINE transposons accounted for 0.53% to 5.75% teleost genomic sequences, and 14 superfamilies were detected. L1, L2, RTE and Rex retrotransposons obtained significant amplification. While LTR displayed low amplification in most teleost with less than 2% of genome coverages, except in zebrafish and stickleback, where LTR reachs 5.58% and 2.51% of genome coverages respectively. And 6 LTR superfamilies (Copia, DIRS, ERV, Gypsy, Ngaro and Pao) were detected in the teleost, and Gypsy exhibits obvious amplication among them. While the SINE represents the weakest ampification types in teleost, only within zebrafish and coelacanth, it represents 3.28% and 5.64% of genome coverages, in the other 7 teleost, it occupies less than 1% of genomes, and tRNA, 5S and MIR families of SINE have a certain degree of amplification in some teleosts. This study shows that the teleost display high diversity and large variation of mobilome, there is a strong correlation with the size variations of genomes and mobilome contents in teleost, mobilome is an important factor in determining the teleost genome size.

Estrogen receptor (esr) mediates the effects of estrogen on the expression of related genes, thereby regulating the growth and reproduction of mammals. To investigate the effect of retrotransposon insertion polymorphism (RIP) of the porcine esr gene on porcine growth performance, retrotransposon insertion polymorphism of the esr gene were predicted by comparative genomics and bioinformatics, and PCR was used to verify the insertion polymorphisms in different porcine breeds. Finally, the correlation analysis between the genotypes and performance of Large White pigs was conducted. The results showed that four retrotransposon polymorphic sites were identified in the esr1 and esr2 genes, which are esr1-SINE- RIP1 located in intron 2 of the esr1 gene, esr1-LINE-RIP2 and RIP3-esr1- SINE located in intron 5 of the gene, and esr2-LINE-RIP located in intron 1 of the esr2 gene, respectively. Among them, insertion of a 287 bp of SINE into intron 2 of the esr1 gene significantly affected (P<0.05) the live back fat thickness and 100 kg body weight back fat thickness of Large White pigs. Moreover, the live back fat thickness and back fat thickness at 100 kg body weight of homozygous with insertion (SINE+/+) was significantly greater than that of heterozygous with insertion (SINE+/-) and homozygous without insertion (SINE-/-). Therefore, esr1-SINE-RIP1 could be used as a molecular marker to assist the selection of deposition traits in Large White pigs.
Animals ; Genotype ; Introns/genetics* ; Phenotype ; Polymorphism, Genetic/genetics* ; Retroelements/genetics* ; Swine/genetics*