Purpose To construct expression vector of Fas extracellular region gene(eFas) ,to express and purify recombination protein and to prepare polyclonal antibody, which have laid a foundation of studying its function. Methods The eFas gene encoding sequence was acquired through overlapping PCR, and pET-22b ( + )/eFas expression vector was constructed. Then this vector was transformed into E. coli Rosetta-gami. Re-combinant protein was expression being induced by IPTG,and was purified using Ni-NTA matrix of affinity chromatograph. The purity of recombination protein was identified by SDS-PAGE. Hereafter, the purified eFas recombinant protein was immunized to New Zealand white rabbit in order to prepare polyclonal antibody. The titer of polyclonal antibody was determined by ELISA. Results The encoding sequence and expression vector of eFas was obtained while the interest protein was mainly expressed in the inclusion body. The eFas fusion protein's expression quantity accounts for more than 30% proportion of total E. coli protein. The eFas protein we obtained was provided with the purity of at least 95 % . Conclusion The successful constrution, expression and purification of FasL fusion protein and preparation of polyclonal antibody will provide some material for further studies of Fas.

Objective To establish a method for detecting the chemical compositions of “decocted first and defoamed” of Ephedra Herba by UPLC-DAD-TOF/MS coupled with HPLC-UV, so as to clarify the difference of chemical constituents among them. Methods The analysis was performed on an ACQUITY UPLC® BEH C18 column (50 mm × 2.1 mm, 1.7 μm ) by UPLC-DAD-TOF/MS with gradient elution. The mobile phase consists of methanol and 0.1% formic acid-water at a flow rate of 0.3 mL/min. The column temperature was 40 ℃. The information of compounds was acquired on positive and negative mode. Similarly, HPLC-UV was applied for measuring the content of alkaloids respectively. The C18 column was used, the mobile phase was acetonitrile-0.1% phosphoric acid (containing 0.05% triethylamine) (99 : 1) at the flow rate of 1 mL/min, the detective wavelength was set up at 210 nm, and the column temperature was 30 ℃. Results There were less content in the upper foam, and the chemical components in the lower liquid and the whole liquid were basically same. Five alkaloids and one carboxylic acid (4-hydroxy-7-methoxyl-2-quinoline carboxylic acid) was identified from all kinds of liquids. However, the content of alkaloids in the upper foam was very low, and the content of three alkaloids in the whole solution was slightly higher than that in the lower liquid. Conclusion The defoamed method may not be related to the chemical compositions of alkaloids, but it still needs further research and verification.

AIM: To investigate the dynamic alteration of cardiac collagen metabolism in mice with acute,chronic myocarditis and dilated cardiomyopathy(DCM).METHODS: BALB/c mice infected with coxsackievirus B_3 were used to establish animal models of acute,chronic myocarditis and dilated cardiomyopathy,while uninfected animals were also prepared and served as controls.After verification of models by histopathological methods and echocardiography,serum concentration of aminoterminal propeptide of type Ⅲ procollagen(PIIINP),aminoterminal propeptide of type Ⅰ procollagen(PINP) and carboxyterminal propeptide of type I procollagen(PICP) in each group of mice were detected by enzyme linked immunosorbent assay(ELISA).The expression of matrix metalloproteinase 1(MMP1) and its tissue inhibitor(TIMP-1) were determined by Western blotting analysis.The MMP-1 activity was also detected.RESULTS: Marked myocardial fibrosis was observed in all groups of CVB_3-infected mice.Reparative fibrosis,promotion of synthesis and degradation of cardiac collagens were presented in heart tissue of acute myocarditis mice. Both reparative and reactive fibrosis,enhanced synthesis and lightened degradation of collagen were present in chronic myocarditis,while reactive fibrosis and excess collagen synthesis were confirmed in DCM.Expression and activity of(MMP-1) was progressively decreased.TIMP-1 showed unchanged.The ratio of MMP-1/TIMP-1 was progressively descended.CONCLUSION: Collagen metabolism was special in different phase of viral heart diseases,which may play different roles in the progression and prognosis of these kinds of disease.

This article described the background,concept,characteristic and objective of the research-based nursing,systematically introducing the main measures including management mechanism, nursing service,nursing staff training,and nursing scientific development.Other areas covered include innovation management mechanism,updating service philosophy,improving nursing staff training,and constructing scientific research platform.

Objective To investigate the therapeutic effects of hemin, an inducer of heme oxygenase, in a rat model of gestational hypertension and explore the possible mechanism. Methods Eighteen pregnant SD rats at day 12 of gestation were randomized equally into gestational hypertension model group, hemin treatment group, and normal pregnancy (control) group. In the former two groups, the rats were subjected to daily nitro-L-arginine methyl ester (L-NAME, 80 mg/kg) gavage since gestational day 14 for 7 consecutive days to induce gestational hypertension; saline was administered in the same manner in the control rats. The rats in hemin group received daily intraperitoneal injection of hemin (30 mg/kg) starting from gestational day 16. HO activity and carboxyhemoglobin (COHb) level in rat placental tissue were detected with spectrophotometric method, and soluble vascular endothelial growth factor receptor-1 (sFIt-1) and vascular endothelial growth factor (VEGF) level in the placental tissue homogenate supernatant were detected using ELSIA. Results At gestational day 20, the blood pressure and 24-h urinary protein were significantly higher in the model group than in the other two groups (P<0.05), and were higher in hemin group than in the control group (P<0.05);HO activity and COHb content in the placenta tissue were the lowest in the model group (P<0.05), and was lower in hemin group than in the control group (P<0.05). The level of sFIt-1 was significantly higher and VEGF level significantly lower in the model group than in the other two groups (P<0.05);sFIt-1 level remained higher and VEGF lower in hemin group than in the control group (P<0.05). Conclusion Hemin can reduce blood pressure and urinary protein in rats with gestational hypertension possibly by up-regulating HO activity, enhancing carbon monoxide production, reducing sFIt-1 and increasing VEGF in the placental tissue.

Objective To investigate the therapeutic effects of hemin, an inducer of heme oxygenase, in a rat model of gestational hypertension and explore the possible mechanism. Methods Eighteen pregnant SD rats at day 12 of gestation were randomized equally into gestational hypertension model group, hemin treatment group, and normal pregnancy (control) group. In the former two groups, the rats were subjected to daily nitro-L-arginine methyl ester (L-NAME, 80 mg/kg) gavage since gestational day 14 for 7 consecutive days to induce gestational hypertension; saline was administered in the same manner in the control rats. The rats in hemin group received daily intraperitoneal injection of hemin (30 mg/kg) starting from gestational day 16. HO activity and carboxyhemoglobin (COHb) level in rat placental tissue were detected with spectrophotometric method, and soluble vascular endothelial growth factor receptor-1 (sFIt-1) and vascular endothelial growth factor (VEGF) level in the placental tissue homogenate supernatant were detected using ELSIA. Results At gestational day 20, the blood pressure and 24-h urinary protein were significantly higher in the model group than in the other two groups (P<0.05), and were higher in hemin group than in the control group (P<0.05);HO activity and COHb content in the placenta tissue were the lowest in the model group (P<0.05), and was lower in hemin group than in the control group (P<0.05). The level of sFIt-1 was significantly higher and VEGF level significantly lower in the model group than in the other two groups (P<0.05);sFIt-1 level remained higher and VEGF lower in hemin group than in the control group (P<0.05). Conclusion Hemin can reduce blood pressure and urinary protein in rats with gestational hypertension possibly by up-regulating HO activity, enhancing carbon monoxide production, reducing sFIt-1 and increasing VEGF in the placental tissue.

OBJECTIVES: Pelvic floor dysfunction (PFD) seriously affects women's physical and mental health. Pregnancy and childbirth are recognized as high-risk factors for PFD, and studies have shown that vaginal microenvironmental disorders can promote the development of pelvic organ prolapse. In this study, we intend to investigate whether the changes in vaginal microecology during pregnancy affect the pelvic floor function and participate in the development of postpartum PFD, and provide a basis for the prevention and treatment of PFD. METHODS: A total of 358 full-term mothers who delivered in Third Xiangya Hospital, Central South University from November 2019 to April 2020 were selected and underwent review 6 to 8 weeks after delivery. The pelvic floor structures were examined using pelvic floor ultrasound, and ultrasound values were measured at rest and at maximum Valsalva maneuver. One hundred and seventy women with PFD were assigned in a PFD group, and 188 women without PFD were assigned in a control group. The clinical data of all mothers were collected, and the clinical data and the results of microecological testing for vaginal secretions after 36 weeks of gestation and before delivery were compared between the 2 groups. The correlation of PFD with leucorrhoea cleanliness, lactobacillus level, bacterial vaginosis (BV), and vulvovaginal candidiasis (VVC) was analyzed, and logistic regression analysis was used to screen for independent risk factors for PFD. RESULTS: The incidences of VVC, BV, Lactobacillus vaginalis deficiency, and leucorrhoea cleanliness ≥III° were all higher in the PFD group than those in the control group (P<0.05). Among them, leukocyte cleanliness ≥III°and lack of Lactobacilli in the vagina were independent risk factors for the development of PFD, while VVC and BV were not independent risk factors for the development of PFD. CONCLUSIONS Postpartum PFD is related to vaginal microecological imbalance in late pregnancy, among which Lactobacillus vaginalis deficiency and leucorrhoea cleanliness ≥III° are independent risk factors for the occurrence of PFD. Therefore, pregnant women with Lactobacillus vaginalis deficiency and leucorrhoea cleanliness ≥III° in late pregnancy should pay attention to the occurrence of postpartum PFD, and early diagnosis and effective intervention of postpartum PFD should be enhanced.
Pregnancy ; Female ; Humans ; Pelvic Floor ; Mothers

Objective: To understand the mechanism of chemotherapy resistance in nasopharyngeal carcinoma under hypoxic conditions through the perspective of protein SUMOylation modification. Methods: Cobalt chloride (CoCl₂) was used to establish the hypoxic model of human nasopharyngeal carcinoma CNE1 cells. Then, the cell cycle was detected by flow cytometry, and the expression level of small ubiquitin-related modifier(SUMO) and cyclin-dependent kinase 6 (CDK6) proteins were detected by western blotting. MTT assay was used to determine the median lethal dose (IC₅₀) of cancer cells against cisplatin, and enzyme-linked immunosorbent assay (ELISA) was used to determine lactate dehydrogenase (LDH) level. Results: The cell cycle of CNE1 induced by hypoxia was arrested in G0/G1 phase.The results of Western blot showed that the protein expression level of CDK6 in CNE1 cells was lower than that in the control group (0.83±0.25 vs. 0.43±0.21, t=14.67, P=0.003). The protein level of conjugated SUMO1 was significantly lower than that in the control group (2.69±0.48 vs. 1.38±0.31, t=17.22, P=0.001), while the level of free SUMO1 protein was significantly higher than that in the control group (2.01±0.43 vs. 2.60±0.59, t=15.45, P=0.002).The LC50 of CNE1 cells in the control group was significantly lower than that in the hypoxic group (29.44 μg/ml vs. 97.72 μg/ml, t=12.79, P=0.001). After CNE1 cells received 50 μg/ml cisplatin for 48 h, the LDH content in the supernatant of the control group was significantly higher than that in the hypoxic group ((541.49±64.59) ng/ml vs. (234.67±41.03) ng/ml, t=11.94, P=0.007)). The apoptosis rate of CNE1 cells in the control group was significantly higher than that in the hypoxic group ((76.64±5.37)% vs. (32.84±4.77) ng/ml, t=8.49, P=0.003)). Conclusion Hypoxia can dissociate the covalent modification of CDK6 and SUMO1, inhibit cell cycle and increase the chemotherapy resistance of nasopharyngeal carcinoma.

Spontaneous rupture of the ovarian artery is very rare and can cause retroperitoneal hemorrhage, which is seriously life-threatening. Herein, we reported a case of massive retroperitoneal hematoma caused by spontaneous rupture of the right ovarian artery during pregnancy and intrauterine fetal death. A 32-year-old woman, gravida 6 para 5, had non-specific right lower abdomen and low back pain in the third trimester. Emergency cesarean section was performed due to the increased pain and decreased fetal heart rate. A huge retroperitoneal hematoma and intrauterine fetal death were found. Then, the abdomen was closed due to unknown source of bleeding and unstable vital signs. Computed tomography scan was conducted to clarify the extent of the retroperitoneal hematoma. Digital subtraction angiography confirmed the rupture of the right ovarian artery. A transcatheter artery embolization was successfully performed to control the bleeding. The patient ultimately recovered well after surgery.
Pregnancy ; Humans ; Female ; Adult ; Rupture, Spontaneous ; Cesarean Section ; Fetal Death ; Arteries

Objective To investigate the effect of sodium-glucose cotransporter 2 inhibitor (empagliflozin, EMPA) on myocardial remodeling in a mouse uremic cardiomyopathy (UCM) model induced by 5/6 nephrectomy, through the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (PKB/AKT)/p65 signaling pathway. Methods The animals were divided into three groups: Sham group (n=6), UCM group (n=8), and UCM＋EMPA group (n=8). A UCM model was established in C57BL/6N mice using the 5/6 nephrectomy. Starting from 5 weeks post-surgery, EMPA or a placebo was administered. After 16 weeks, blood pressure, serum creatinine, blood urea nitrogen, 24-hour urine glucose and urine sodium were measured. Cardiac structure and function were assessed by echocardiography. Hematoxylin-eosin (HE) staining and Masson trichrome staining were used to observe pathological changes in the heart and kidneys. Wheat germ agglutinin (WGA) staining was used to evaluate myocardial hypertrophy. The real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of myocardial hypertrophy- and fibrosis-related mRNAs. Western blotting was used to detect the expression levels of PI3K, AKT and p65 in myocardial tissues. Results After 16 weeks, UCM group exhibited significantly higher blood pressure, serum creatinine, blood urea nitrogen than sham group (P＜0.01); UCM＋EMPA group exhibited lower blood pressure, serum creatinine, blood urea nitrogen, and higher 24 h urine sodium and glucose than UCM group (P＜0.05). Echocardiographic results showed ventricular remodeling in the UCM group, evidenced by left ventricular wall thickening, left ventricular enlargement, increased left ventricular mass, and decreased systolic function (P＜0.05); ventricular remodeling was alleviated (P＜0.05), though there was no significant improvement in systolic function in UCM＋EMPA group. HE and Masson stainings revealed myocardial degeneration, necrosis, and interstitial fibrosis in UCM group (P＜0.01); the myocardial pathology improved with reduced collagen deposition in UCM＋EMPA group (P＜0.01). WGA staining confirmed myocardial hypertrophy in UCM group (P＜0.01), while myocardial hypertrophy was alleviated in UCM＋EMPA group (P＜0.01). RT-qPCR results showed myocardial hypertrophy- and fibrosis-related genes (NPPA, NPPB, MYH7, COL1A1, COL3A1, TGF-β1) were upregulated in UCM group (P＜0.05), but downregulated in UCM＋EMPA group. Western blotting showed PI3K, p-AKT/AKT ratio, and p-p65/p65 ratio were increased in UCM group, but decreased in UCM＋EMPA group (P＜0.05). Conclusion EMPA can improve myocardial hypertrophy and fibrosis in the UCM mouse model, and it may play the role through inhibiting the PI3K/AKT/p65 signaling pathway.