Objective To explore whether Cdc42 is an independent influence factor in regulating the migration of A549 cells by using quantitative measurement method,and to verify the effectiveness of an automatic measurement and calculation method for in vitro cell migration assay.Methods Cdc42 was overexpressed in human lung adenocarcinoma A549 cell line,and then in vitro scratch assay was applied to evaluate the migration of the cells.Different methods were used to measure the images acquired at different time points for quantitative analysis.Accuracy and repeatability of different measurement methods were analyzed.Results The results showed that overexpression of Cdc42 alone significantly advanced the migration of A549 cells (P＜0.05).The new method is efficient,accurate and reproducible as compared to manual measurement,and has significant advantages in target recognition and noise removal as compared to the existing measurement method in Image Pro Plus 6.0.Conclusions Over expression of Cdc42 significantly increased A549 cell migration ability in vitro.The new method can realize the automatic quantitative analysis of cell migration in vitro.

Objective To explore an approach to identifying the plant source of Flos Lonicerae and Lonicera confusa rapidly, precisely, and efficiently. Methods Using LM and SEM to observe and compare the characters of leaf epidermis of the five species of Lonicer L. including L. japonica, L. confuse, L. hypoglauca, L. dasystyla, and L. macrantha. Results There are obvious differences in leaf epidermis among the five species of Lonicera L. under LM and SEM. Conclusion The characteristics of leaf epidermis can be used to identify the plant source of Flos Lonicerae and L. confusa. According to their characteristics under LM and SEM, the retrieval keys to the five species of Lonicera L. have been compiled.

Hormone replacement therapy (HRT), involving giving sex steroid hormones such as estrogen alone or with a progestogen, is widely used in postmenopausal women.HRT helps to relieve menopausal symptoms and has also been shown to prevent osteoporosis.Although most observational studies have showed that HRT can reduce the risks of cardio-cerebrovascular diseases, the subsequent randomized controlled trials were inconsistent with the results.This article reviews the relationship between HRT and stroke from drug type, route of administration, estrogen dosage, and initiation time.

Fluid-attenuated inversion recovery vascular hyperintensity (FVH) is a common magnetic resonance imaging findings in acute ischemic stroke due to severe stenosis or occlusion of large cerebral arteries.This article reviews the applications and related research of FVH in patients with acute ischemic stroke.

Vertebral artery hypoplasia is a congenital vessel variation. Its incidence is from 1. 9 to 26. 5% . In recent years, studies have shown that vertebral artery hypoplasia may be a potential risk factor for posterior circulation infarction, especialy when it coexists with other cerebrovascular risk factors. Vertebral artery hypoplasia may also cause regional hypoperfusion and complex neurovascular regulation, and it also has a certaln link with migralne.

Adult ; Epilepsy ; genetics ; Humans ; Hyperhomocysteinemia ; genetics ; Hypogonadism ; genetics ; Intellectual Disability ; genetics ; Male ; White Matter ; pathology

Objective To analyze the platelet activating factor acetylhydrolase ( PAF-AH) activity of a gene product encoded by LA2144 gene of Leptospira interrogans ( L. interrogans) , to investigate the ex-pression and secretion of LA2144 protein in various cell cultures and to further understand its function in in-ducing internal hemorrhage in an animal model. Methods The DNA sample containing LA2144 gene was extracted from L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai and used as the template for gene cloning by PCR. The LA2144 gene without the signal sequence coding region was amplified by PCR and inserted into a prokaryotic expression construct for the protein expression. The expressed recombinant protein, rLep-PAF-AH, was purified by Ni-NTA affinity chromatography. Spectrophotometry was used to measure the hydrolytic activity, hydrolytic efficiency, Km and Kcat values of the rLep-PAF-AH protein in hydrolyzing PAF substrate. Real-time fluorescent quantitative RT-PCR ( qRT-PCR) and Western blot assay were performed to measure the expression of LA2144 gene at mRNA and protein levels in human umbilical vein endothelial cells (HUVEC), human monocytes (THP-1) and murine macrophages (J774A. 1) with L. interrogans strain Lai infection, respectively. Each syrian hamster was intravenously injected with 100 μg of LPS-free rLep-PAF-AH for two times. Hemorrhage in the lungs, livers and kidneys were observed in three days after the injection. Results The constructed prokaryotic expression system for LA2144 gene of L. inter-rogans strain Lai could highly express the rLep-PAF-AH upon the induction of IPTG. The purified rLep-PAF-AH showed high purity with a single protein band in gel as indicated by SDS-PAGE. The efficiency of 5 μg of rLep-PAF-AH in hydrolyzing PAF substrate was 26. 6 U/L with a Km value of 82. 79 μmol/L and a Kcat value of 0. 24 S-1 . The expression of Lep-PAF-AH at mRNA level in HUVEC, THP-1 and J774A. 1 cells were significantly elevated after co-culture with L. interrogans strain Lai for 1 or 2 hours (P<0. 05). A large amount of Lep-PAF-AH were detected in the supernatants from co-cultures of L. interrogans strain Lai with the three cell lines, but not from the culture of the spirochete in EMJH medium. The signs of hemor-rhage were observed in the lung of hamsters injected with rLep-PAF-AH, but not in tissue samples from liver and kidney. Conclusion The LA2144 gene product was characterized by a stronger PAF-AH activity. The expression of LA2144 gene at mRNA and protein levels in various cell lines were enhanced during L. interro-gans infection. Moreover, the rLep-PAF-AH could induce the pulmonary hemorrhage in hamsters. This stud-y indicated that the protein encoded by LA2144 gene was an important virulence factor causing hemorrhage in hosts during L. interrogans infection.

BACKGROUND:Previous studies have shown that kainic acid injected into hippocampus can significantly upregulate the expression of excitatory KA1 subunit of the kainate receptor in the hippocampus, and endoplasmic reticulum stress markers, phosphorylation of the alpha subunit of eukaryotic initiation factor 2, accompanied by celldeath. OBJECTIVE:To explore the mechanism of endoplasmic reticulum stress after kainic acid is injected into the hippocampus.METHODS:0.15 nmol kainic acid was injected into the hippocampal CA1 region of 32 adult male Kunming mice, the injection time was 60 seconds. At different time points (1, 2, 3, 4, 5, 6, 8 and 12 hours) after kainic acid was injected, the Bederson score analysis was performed, and then the brain was harvested after cerebral perfusion. FJB staining of brain sections and immunofluorescence double labeled observation were also performed. RESULTS AND CONCLUSION:(1) At 3, 4, 5, 6, 8 hours after kainic acid injection, Bederson score showed severe injury of central nervous system function, and FJB staining showed the increased of celldeath in the hippocampus (P<0.05);At 1, 2, 12 hours after injection, Bederson score showed no obvious injury of central nervous system function, and FJB staining showed unobvious celldeath in the hippocampus (P>0.05). (2) According to the results of FJB staining, the brain sections were selected at 3, 8 hours for immunohistochemistry. The expressionlevels of KA1 receptors and endoplasmic reticulum stress marker P-eIF2αwere up-regulated at the same time after kainic acid was injected into hippocampus. Two single-staining KA1 and P-eIF2αimmunofluorescence images were synthesized into one over-lapped double-stained image, and two images overlapped, indicating that the up-regulated expression of KA1 and endoplasmic reticulum stress occurred in the same nerve cells. Kainic acid first up-regulated the excitatory receptor KA1 expression, which may cause cellendoplasmic reticulum dysfunction and result in the endoplasmic reticulum stress response, further promoting neuronal celldeath.

Objective To investigate the impact of hepatectomy combined with splenectomy on hepatitis B virus (HBV) reactivation in patients with hepatocellular carcinoma (HCC) ≤5 cm and with hypersplenism.Methods This is a retrospective case-control study on 167 patients with HCC ≤5 cm and with hypersplenism who underwent hepatectomy combined with splenectomy at the Shengli Oilfield Central Hospital between May 2008 and June 2015.64 patients underwent hepatectomy combined with splenectomy,and 103 patients hepatectomy alone.The patients were assigned to the hepatectomy combined with splenectomy group (the combined group,n =61) or the hepatectomy alone group (the control group,n =61) using propensity score matching (PSM).Logistic regression was used to evaluate the relative clinical factors associated with HBV reactivation.The stratified Chi-squared test was utilized to determine the impact of the surgical procedure and preoperative anti-viral therapy on postoperative hepatitis B virus reactivation of these patients.Results The serum PLT level,Child-Pugh grading,tumor diameter and surgical procedures were shown to be independent risk factors associated with postoperative HBV reactivation (P ＜ 0.05).To study the impact of preoperative anti-viral therapy on postoperative HBV reactivation:-the incidence of HBV reactivation was higher in the control group than in the combined group (19.7％ vs.6.6％,P ＜ 0.05).In the combined group,there was no significant difference between patients who received anti-viral therapy and those who were treatment-naive (5.3％ vs.7.1％,P ＞0.05).In the control group,a higher incidence of HBV reactivation was found in patients with treatment-na(i)ve than in patients who received anti-viral therapy (26.1 ％ vs.0,P ＜ 0.05).For the patients who received anti-viral therapy,there was no significant difference between the combined group and the control group (5.3％ vs.0,P ＞ 0.05).In patients with treatment-na(i)ve,a higher incidence of HBV reactivation was observed in the control group than the combined group (26.1％ vs.7.1％,P ＜ 0.05).Conclusions In patients who were not treated with antiviral therapy,hepatectomy combined with splenectomy decreased the incidence of postoperative HBV reactivation in patient with HCC ≤5 cm and with hypersplenism.For the patients who received preoperative anti-viral therapy,the incidence of postoperative HBV reactivation was not decreased with hepatectomy combined with splenectomy.