OBJECTIVE:To investigate the effects of Qigong pills formula on uterine artery blood flow and endometrial recep-tivity of patients with polycystic ovary syndrome(PCOS)with infertility. METHODS:A total of 86 PCOS patients with infertility in our hospital during Jan.-Dec. 2016 were divided into control group and observation group according to random number table, with 43 cases in each group. Control group was given western medicine pituitary down-regulation therapy. Observation group was additionally given adjuvant therapy of Qigong pills formula,a dose per day,decocted with water to 400 mL,morning and night un-til artificial fertilization stopped,on the basis of control group. The uterus ultrasonic indexes and endometrial receptivity were com-pared between 2 groups before treatment and 3 months after treatment. The occurrence of ovulation,pregnancy and ADR was re-corded. RESULTS:Before treatment,there was no statistical significance in uterus ultrasonic indexes and endometrial receptivity in-dexes between 2 groups (P>0.05). Three months after treatment,endometrial thickness of 2 groups were increased significantly, while endometrial spiral arterial pulsatility indexes and resistance indexes were decreased significantly;the levels ofαvβ3 and GLUT4 were increased significantly;observation group was significantly better than control group,with statistical significance (P<0.05). The ovulation rate of observation group was 83.72％,which was significantly higher than 62.79％ of control group,with statistical significance(P<0.05). There was no statistical significance in pregnancy rate(27.91％ vs. 18.60％)between 2 groups(P>0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:The adjuvant therapy of Qigong pills formula help to regulate the pa-rameters of uterine artery blood flow in PCOS patients with infertility,and strengthen endometrial receptivity so as to increase ovu-lation rate.

Objective To investigate the function of tripartite motif protein 22 (TRIM22) and the interaction with eukaryotic translation initiation factor-4E (eIF4E) in the differentiation of NB4 cells, one kind of acute promyelocytic leukemia cells, which elucidates the mechanism of TRIM22 targeting to regulate eIF4E.Methods The model of NB4 cells inducing differentiation was established in vitro.The expression changes of gene and protein of TRIM22 and eIF4E were detected by using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.In addition, the effect on cell function and protein expression level of eIF4E after adopting electroporation technology to depress or over-express TRIM22 was detected by CCK-8 and flow cytometry.Finally, the interaction of TRIM22 and eIF4E was verified by using co-immunoprecipitation (CO-IP).Results The mRNA relative expression level of TRIM22 was gradually increasing from 1.01±0.15 to 30.98±2.79 (F=280.700, P=0.000), and the protein relative expression level was gradually increasing from 0.22±0.03 to 0.51±0.05 (F=51.430, P=0.000) after the all-trans-retinoic acid (ATRA) induction for NB4 cell.However, the mRNA relative expression level of eIF4E was gradually decreasing from 1.01±0.09 to 0.47±0.06 (F=20.520, P=0.000), with the same trend, the protein relative expression level was gradually decreasing from 0.97±0.02 to 0.64±0.09 (F=14.700, P=0.001).The expression level of PE-CD11b in the TRIM22 over-expression group with ATRA detected by flow cytometry [(78.80±2.00)%] was higher than that in the transfection group of empty vetor with ATRA [(58.70±2.70)%] (t=9.535, P=0.000) and the cotransfection group with ATRA [(61.60±3.80)%] (t=8.187, P=0.000).Meanwhile, the protein level of eIF4E changed reversely after over-expressing the gene level of TRIM22 (t=4.985, P=0.007).The CO-IP experiment was used to verify the interaction of TRIM22 and eIF4E.ConclusionTRIM22 is able to promote the cell differentiation during the process of NB4 cells differentiation.Furthermore, eIF4E is a target of TRIM22 for binding with, which plays an important role in depressing the expression of eIF4E.

Objective: An innovative technique was established to rapidly construct various cell lines that could be induced to express multiple influenza A virus (IAV) proteins. Method: The HA protein genes of multiple IAVs were cloned into the Cumate-induced expression system which was positioned between two PiggyBac transposon sites. These HA plasmids were transfected into the HEK293A cell line with the PiggyBac transposase plasmids. The transfected cells were screened with puromycin, and after that the corresponding virus proteins were induced with Cumate. Results: The results of flow cytometry and Western blotting showed that the virus proteins were expressed in most of the cells in corresponding lines after the induction of Cumate. Conclusion Cell lines which were inducible to express IVA HA protein can be efficiently constructed by using the PiggyBac transposon system.

In order to study the functions of the HSPs (Heat shock proteins) of Eimeria tenella, we cloned a novel gene (which designated EtHSP) coding HSP of Eimeria tenella by RT-PCR and RACE (Rapid-amplification of cDNA ends). The full-length cDNA sequence of EtHSP was 1802 bp, containing a 1455 bp ORF (Open reading frame) (GenBank Accession No. FJ911605) encoding a deduced protein of 484 amino acids. Real-time PCR revealed that the mRNA level of EtHSP was much higher in sporozoites of E. tenella than other developmental stages (unsporulated oocysts, sporulated oocysts and merozoites). We constructed the recombinant plasmids pET28a(+)-EtHSP, then transformed it into E. coli BL21(DE3) for expression. SDS-PAGE indicated that the fusion protein was expressed in included bodies, with peak expression 6 h after induction by IPTG Western blotting revealed that the protein was specifically recognized by polyclonal antibodies against E. tenella, showing that the fusion protein was native antigen.
Animals ; Chickens ; Eimeria tenella ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Heat-Shock Proteins ; genetics ; immunology ; metabolism ; Inclusion Bodies ; metabolism ; Male ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Rabbits ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Sequence Analysis, Protein

Objective: To evaluate the effect of compound electrolyte injection on phosphatidylserine (PS) exposure in erythrocytes after blood salvage-retransfusion in dogs. Methods: Twenty healthy mongrel dogs, weighing 10-15 kg, aged 3-5 weeks, were divided into 2 groups (n=10 each) using a random number table method: normal saline group (group NS) and compound electrolyte injection group (group MEI). The process of intraoperative blood salvage-retransfusion was simulated in both groups: femoral vein was cannulated for blood withdrawal until the volume of blood lost was 400 ml, and the shed blood was salvaged by a blood recovery machine.The washing solution was normal saline in group NS and compound electrolyte injection in group MEI.The erythrocytes were retransfused after being labeled with fluorescein isothiocyanate.Blood samples were obtained before and after blood salvage for determination of erythrocyte ATP content by enzyme-linked immunosorbent assay.Blood samples were obtained at 24, 48 and 72 h after blood retransfusion, and the PS exposure rate of the salvaged erythrocytes was determined by flow cytometry.The spleen was taken at 72 h after retransfusion to detect the phagocytosis rate of salvaged erythrocytes by monocytes. Results: There was no significant difference in ATP content before and after blood salvage between the two groups (P>0.05). Compared with NS group, the PS exposure rate of the salvaged erythrocytes at each time point after retransfusion and phagocytosis rate of salvaged erythrocytes by monocytes were significantly decreased in MEI group (P<0.05). Conclusion Compound electrolyte injection as a washing solution for intraoperative blood salvage can reduce the PS exposure in salvaged erythrocytes and is helpful in prolonging the lifespan of erythrocytes after retransfusion in dogs.

【Objective】 To confirm the role of Wnt signaling pathway in the occurrence and development of gastric cancer （GC）， establish a prognostic model composed of Wnt pathway related genes， and then evaluate the predictive value of the model. 【Methods】 We downloaded the gene expression data and survival data of GC in TCGA database， and used GSEA enrichment analysis to verify the enrichment of Wnt pathway in GC and para-cancer samples. In this study， univariable COX regression analysis and survival curve analysis were used to select the prognosis-related genes of GC. Then the multivariate COX proportional hazard regression model was used to obtain the prognostic model of Wnt signaling pathway related genes. Then， receiver operating characteristic （ROC） curve and forest plot were used to verify the clinical predictive value of the model. The model was then validated in GEO external database. Finally， by utilizing quantitative real-time PCR （qPCR）， we detected the expressions of Wnt signaling pathway related genes in 8 pairs of clinical GC and para-cancer samples. 【Results】 We downloaded 32 samples of normal para-cancer samples and 375 cancer samples and their corresponding clinical data. GSEA enrichment showed that compared with normal samples， Wnt pathway was significantly enriched in GC samples （P<0.05）. The results of univariate COX analysis showed that 13 Wnt pathway genes were closely related to the prognosis of GC patients. Multivariate COX determined that the model was multiplied and accumulated by ETV2， SERPINE1， CPZ， VPS35 and IGFBP1 and their corresponding coefficient β. The survival curve and ROC curve showed that the model could accurately predict the prognosis of GC patients， and the 1-year， 3-year， and 5-year areas under the curve （AUC） were 68.0%， 69.4% and 78.5%， respectively. Clinical univariate and multivariate COX analyses showed that the model could become an independent prognostic factor other than TNM system of GC. The external data set （GSE84437） validation results of GC showed that the model could better predict the prognosis of GC patients. qPCR results indicated that ETV2， SERPINE1， CPZ， VPS35 and IGFBP1 expressions were upregulated in GC samples compared with para-cancer samples. 【Conclusion】 This study further confirmed that Wnt pathway plays an important role in the progress of GC from the perspective of bioinformatics， and we have established a prognosis-related risk model， providing a new perspective for clinical genetic testing， targeted therapy and individualized therapy.