Total flavonoids of Dracocephalum moldavica L. inhibit oxidative stress against H9c2 cell ischemia-reperfusion injury via VEGF-B/AMPK pathway / 药学学报

Total flavonoids of Dracocephalum moldavica L. (TFDM) is an effective component extracted and isolated from the traditional Uighur medicinal herb Cymbidium fragrans. Cymbidium fragrans has the effects of tonifying the heart and brain, promoting blood circulation and resolving blood stasis, and has been widely used in the treatment of cardiovascular and cerebrovascular diseases for a long time. The purpose of this study was to determine the effect of total flavonoids from Cymbidium fragrans on hypoxia/re-oxygenation (H/R) injury in H9c2 (rat cardiomyocytes) cells and its mechanism. A model (H/R) of hypoxia/re-oxygenation injury in H9c2 cells was established using hypoxia and glucose deprivation for 9 h combined with re-oxygenation and rehydration for 2 h to simulate myocardial ischemia-reperfusion injury. The effects of total flavonoids from Cymbidium fragrans on cell viability, markers of myocardial cell damage, oxidative stress levels, and reactive oxygen radical (ROS) content were investigated, Western blot was used to detect the expression of vascular endothelial growth factor B (VEGF-B) and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) pathway related proteins. The results showed that the total flavonoids of Cymbidium fragrans significantly increased the viability of myocardial cells after H/R injury, and decreased the content of lactate dehydrogenase (LDH) and creatine kinase isozyme (CK-MB) in the cell supernatant. It significantly reduced malondialdehyde (MDA), increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and decreased intracellular ROS and nitric oxide (NO) content. Western blot analysis showed that the total flavonoids of Cymbidium fragrans decreased Bax levels in H9c2 cells damaged by H/R and increased Bcl-2 expression. Total flavones of Cymbidium fragrans upregulate VEGF-B/AMPK pathway related proteins VEGF-B, vascular endothelial growth factor receptor 1 (VEGFR-1), neuropilin 1 (NRP-1), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), phosphorylated adenosine monophosphate activated protein (p-AMPK) and phospho mechanistic target of rapamycin (p-MTOR) levels. The above research results indicate that the total flavonoids of Cymbidium can significantly reduce the H/R injury of myocardial cells, which may be related to the upregulation of VEGF-B/AMPK pathway and inhibition of oxidative stress response.

Expression and localization of vascular endothelial growth factor-B in diencephalon and brainstem related tissues of the Yak’s brain / 解剖学报

Objective To explore the expression and distribution characteristics of vascular endothelial growth factor-B(VEGF-B) in diencephalon and brainstem of Yak’s brain tissues, and to investigate the associations between its expression and hypoxia adaptation. Methods Five healthy yaks were selected, and the brain tissues were divided and collected according to the gross anatomical structure of the brain, including pituitary, thalamus, hypothalamus, oblongata and pons. The characteristics of expression and location of VEGF-B in different regions of Yak’s brain tissues were detected by Real-time PCR, Western blotting and immunohistochemical techniques. Results The results showed that the highest expression level of VEGF-B mRNA of yak brain tissue was in the pituitary, and the content was significantly higher than that found in other parts of the brain(P<0. 05). Following the expressions were in the hypothalamus, thalamus and medulla oblongata, while the lowest expression level was in pons. The expression level of VEGF-B protein in Yak’s brain tissue was similar to the mRNA expression level except that the thalamus was higher than that of hypothalamus. The result of immunohistochemistry showed that VEGF-B protein-positive substances were mainly distributed in the cytoplasm of various types of cells. Among them, the positive staining of VEGF-B was mainly concentrated in eosinophils of pituitary. The positive staining of VEGF-B was mainly concentrated in pleomorphic cells of thalamus and hypothalamus. The distribution of VEGF-B protein-positive substances were mainly focused in nerve cell body of medulla oblongata and pons. Conclusion VEGF-B protein is expressed in both diencephalon and brainstem of yak, which may be closely related to its functions of anti-apoptosis, "survival factor" and angiogenesis. However, the specific mechanism of its neuroprotective effect on Yak brain under hypoxic environment needs to be further studied. The difference of expression in different regions may be related to the tissue specificity and function in different regions of the brain.

Dermatofibrosarcoma protuberans with unusual presentation in vulva / Dermatofibrosarcoma protuberans con presentación inusual en vulva

Abstract Gynecological sarcomas are uncommon and their location in the vulva and vagina has an incidence of 5% of all malignant neoplasms of the female genital tract. We present the case of a 54-year-old patient with a diagnosis of dermatofibrosarcoma protuberans in the vulva, an infrequent pathology, with less than 60 cases reported worldwide in this anatomical location. Clinically it has a locally aggressive behavior, due to the proliferation of spindle cells with pleomorphism and frequent figures of mitosis that infiltrate the reticular dermis and subcutaneous cellular tissue, giving rise to tumor lesions of variable size and with high rates of local recurrence. The treatment of first choice is surgical excision of the tumor with Mohs micrographic surgery or other surgical techniques for complete evaluation of the circumferential and deep peripheral margin. However, the identification of carcinogenesis mechanis ms where the chromosomal translocation t (17; 22) (q22; q13) is recognized, forming the COL1A1-PDGFB fusion gene, which participates in stimulating tumor cell proliferation, allowing treatment with tyrosine kinase inhibitors such as imatinib for neoadjuvant therapy of surgically unresectable tumors and local recurrences.

Resumen Los sarcomas ginecológicos son infrecuentes y la localización de estos en vulva y vagina tienen una incidencia del 5% de todas las neoplasias malignas del tracto genital femenino. Presentamos el caso de una paciente de 54 años con diagnóstico de dermatofibrosarcoma protuberans en vulva, una patología infrecuente, con menos de 60 casos reportados a nivel mundial en esta localización anatómica. Clínicamente tiene un comportamiento localmente agresivo, debido a la proliferación de células fusiformes con pleomorfismo y frecuentes figuras de mitosis que infiltran la dermis reticular y tejido celular subcutáneo, dando origen a lesiones tumorales de tamaño variable y con altas tasas de recurrencia local. El tratamiento en primera elección es la escisión quirúrgica del tumor con cirugía micrográfica de Mohs u otras técnicas quirúrgicas para evaluación completa del margen periférico circunferencial y profundo. Sin embargo, la identificación de mecanismos de carcinogénesis donde se reconoce la translocación cromosómica t (17; 22) (q22; q13), formando al gen de fusión COL1A1-PDGFB, el cual participa estimulando la proliferación celular tumoral, ha permitido la utilización de los inhibidores de la tirosina quinasa como el imatinib para la realización de terapia neoadyuvante en casos de tumores irresecables quirúrgicamente y en recurrencias locales.

Serum levels of complement B and D in patients with diabetic peripheral neuropathy and their correlation / 中国医师杂志

Objective:To investigate the serum level and significance of complement factor B (CFB) and complement factor D (CFD) in patients with type 2 diabetes mellitus (T2DM) and diabetic peripheral neuropathy (DPN).Methods:From October 2019 to October 2020, 110 patients with T2DM in the endocrinology department of Affiliated Hospital of Jining Medical College were divided into DPN group ( n=60) and simple T2DM group ( n=50) according to whether or not DPN was combined. In addition, 52 cases of physical examination population in the physical examination center in the same period were selected as the normal control group ( n=52). The serum levels of CFB, CFD and tumor necrosis factor-α (TNF-α) were measured by enzyme linked immunosorbent assay (ELISA). The correlation between CFB, CFD and clinical indexes was analyzed, and the influencing factors of DPN were analyzed by logistic regression. Results:The serum levels of CFB and CFD in DPN group were higher than those in T2DM group and normal control group [CFB: (845.43±101.10)μg/ml vs (792.19±116.59)μg/ml, (739.20±123.43)μg/ml, P<0.05], [CFD: (491.71±41.03)mg/L vs (467.58±45.16)mg/L, (445.16±50.47)mg/L, P<0. 05]. Pearson correlation analysis showed that the serum level of CFB was positively correlated with glycosylated hemoglobin (HbA 1c), fasting plasma glucose (FPG) and TNF-α (all P<0.05) and negatively correlated with triiodothyronine (FT3) and total bilirubin (TBIL) (all P<0.05). Serum CFD level was positively correlated with systolic blood pressure, HbA 1c, FPG and TNF-α (all P<0.05), but negatively correlated with FT3 and TBIL (all P<0.05). Logistic regression analysis showed that CFB and CFD were still influential factors for the occurrence and development of DPN after excluding confounding factors such as systolic blood pressure, HbA 1c, FPG, FT3, DBIL, TBIL and TNF-α. Conclusions:(1) Serum CFB and CFD levels were significantly increased in DPN patients, suggesting that CFB and CFD may be involved in the occurrence and development of DPN. (2) Serum TNF-α level was significantly increased in DPN patients, confirming the role of TNF-α in the pathogenesis of DPN.

Study of the protective effect of total flavonoids of Dracocephalum moldavica L.On doxorubicin induced injury in HUVECs based on VEGF-B/AMPKa pathway / 中国药理学通报

Aim To investigate the protective effect of TFDM on doxorubicin-induced endothelial cell injury and its mechanism.Methods Cell viability was detected by CCK-8 assay.Cell morphology was observed by microscope.The changes of LDH, SOD and mitochondrial membrane potential were detected by kit method.Cell migration was detected by Transwell assay; Endothelial dysfunction and VEGF-B/AMPKa pathway related protein expression were detected by Western blot.Results Compared with model group, TFDM significantly increased cell viability, improved the morphologic changes of HUVEC induced by DOX, decreased LDH leakage, increased SOD activity, increased mitochondrial membrane potential, promoted endothelial cell migration, and inhibited endothelial cell injury.The results of Western blot showed that com pared with control group TFDM increased the expression levels of non-receptor tyrosine kinase ( Src) and focal adhesion kinase (FAK) .increased the phosphorylation level of eNOS, and decreased the expression level of ET-1 protein, thereby inhibiting endothelial dysfunction.The protein expression levels of VEGF-B, NRP1 , VEGFR1 and the ratio of p-AMPKa/AMPKa significantly increased in the administration group.Conclusion TFDM may inhibit doxorubicin-induced endothelial cell injury by activating VEGF-B/AMPKa pathway.

The change of serum complement factor B and its clinical implication in children with primary IgA nephropathy / 中华实用儿科临床杂志

Objective:To detect serum level of complement factor B (CFB), and to explore its correlations with clinical parameters and prognosis in children with primary IgA nephropathy (IgAN).Methods:A total of 204 children with primary IgAN confirmed by kidney biopsy in the Department of Nephrology of the First Affiliated Hospital of Zhengzhou University from December 2014 to April 2017 were included in IgAN group.During the same period, 84 healthy children were included in healthy control group.Their mean age was (11.0±3.5) years and (10.9±3.2) years, respectively.Patients in IgAN group were divided into low CFB group (102 cases) and high CFB group (102 cases) according to the medium serum level of CFB measured by enzyme-linked immunosorbent assay. Spearman′ s coefficient was employed to analyze correlation amongst various parameters.Multivariable-adjusted Cox proportional ha-zards models were used to evaluate the relationship between serum CFB level and prognosis in children with IgAN. Results:Serum CFB levels were significantly higher in IgAN group than that in healthy control group [290.9 (186.2-453.9) mg/L vs.218.9 (155.0-321.3) mg/L, Z=-3.372, P=0.001]. Serum levels of CFB were negatively correlated with serum albumin ( r=-0.388, P<0.001) and estimated glomerular filtration rate ( r=-0.416, P<0.001), but positively correlated with serum creatinine ( r=0.305, P<0.001) and 24 h urinary protein ( r=0.456, P<0.001) in IgAN group.The incidences of crescents (C1-2) (70.6% vs.29.4%, χ2=34.588, P<0.001) and C 3 deposition (+ + -+ + + ) (63.7% vs.44.1%, χ2=7.892, P=0.005) were significantly higher in high CFB group than those in low CFB group. Kaplan- Meier analysis showed that high CFB levels predicted worse renal outcome in pediatric IgAN patients ( χ2=17.509, P<0.001). Multivariate Cox regression analysis showed that the high CFB level was the independent risk factor for the poor renal outcome ( HR=2.517, 95% CI: 1.284-4.932, P=0.007). Conclusions:High serum levels of CFB are associated with decreased renal function, increased urinary protein excretion, crescentic formation and poor renal outcome in pediatric IgAN patients.

Effects of Shuangdan Mingmu capsule on the expressions of VEGF in the retina of diabetic rat models / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the effects of Shuangdan Mingmu capsule on the expressions of vascular endothelial growth factor-a(VEGF-a), VEGF-b, VEGF-c in the retina of a diabetic rat model. <p>METHODS: Forty male SD rats were divided into Group A(normal group), Group B(model group), Group C(Shuangdan Mingmu group)and Group D(positive control group)10 rats(20 eyes)in each group. A rat model of diabetic retinopathy was established by one-time tail vein injection with STZ(50mg/kg). After modeling for 1wk, the rats were given medicine by gavage. After gavage for 4wk rats were sacrificed, and the expressions of VEGF-a, VEGF-b, VEGF-c in the retina tissues were detected by immunohistochemical method. <p>RESULTS: After gavage for 4wk the average gray values of VEGF-a, VEGF-b and VEGF-c protein in the retina of model group, Shuangdan Mingmu group and positive control group were lower than those of the normal group, and the average optical density were higher than those of the normal group. There was a significant difference between the model group and the normal group(<i>P</i><0.01). The average gray values of VEGF-a, VEGF-b and VEGF-c expression in Shuangdan Mingmu group and positive control group were higher than those in the model group(<i>P</i><0.05)and the average optical density value were lower than those in the model group.(<i>P</i><0.01). <p>CONCLUSION: Shuangdan Mingmu capsule could significantly reduce the expressions of VEGF-a, VEGF-b,VEGF-c in the retina and had a certain protective effect on the retina of rats in the diabetic retinopathy model.

Clinical application evaluation of serum C1q and other complement factors in the diagnosis and prediction of preeclampsia / 中华检验医学杂志

Objective To observe the levels of serum complement C 1q, C3, C4 and factor B in different phases during normal pregnancy;To evaluate the diagnostic value and the predictive value of serum complement C1q, C3, C4 and factor B in preeclampsia (PE).Methods Three groups of subjectes were enrolled from January 2017 to March 2018 in Department of Obstetrics and Gynecology , Peking University Third Hospital.(1) 30 pregnant women in each group at 8-14 weeks, 20-26 weeks and 28-36 weeks were retrospectively selected , and the serum levels of complement C 1q, C3, C4 and B factors were measured and compared.(2)Selecting 17 cases of early-onset mild PE, 47 cases of early-onset severe PE, 24 cases of late-onset mild PE, 27 cases of late-onset severe PE, and 30 normal pregnant cases of the same gestational stage as early-onset /late-onset controls , through ANOVA analysis and comparison between two groups , this study evaluated the diagnostic value of serum complement C 1q, C3, C4 and factor B in PE.(3)To evaluate the predictive effect in PE, it analyzed serum C1q and factor B levels of pregnant women at 20-26 gestation weeks through prospective nested case-control study of 214 cases.Results The levels of serum C1q remained stable in the whole pregnancy .The levels of C3 and factor B increased at the early stage of pregnancy and remained stable after the middle stage .C4 increased early in pregnancy and then remained stable.Compared with the control group , the levels of serum C1q in all four types of PE patients were significantly decreased ( median: 169 mg/L, 161 mg/L, 165 mg/L, 163 mg/L;early-onset, late-onset control group:187 mg/L, 194 mg/L;U=130.500, 426.500, 159.500, 130.500, all P<0.05).Serum C3 levels of all the other three types of PE patients were significantly lower than those of the control groups (median:1170 mg/L, 1323 mg/L, 1223 mg/L;early-onset, late-onset control groups: 1438 mg/L, 1434 mg/L;U =379.000, 246.000, 160.000, all P <0.05 ), except for the early-onset mild PE (1275 mg/L).Serum C4 levels of patients with early/late onset severe PE were significantly lower than those of the control groups ( median: 140 mg/L, 142 mg/L;early-onset, late-onset control groups:223 mg/L, 235 mg/L;U =329.500, 136.500, both P <0.001 ) .Serum factor B levels showed no statistical difference among 3 early on-set groups or among 3 late on-set groups ( early-onset group median:332 mg/L,318 mg/L,early-onset control group 312 mg/L;late-onset group median:316 mg/L,314 mg/L, late-onset group 303 mg/L;χ2 =5.990, 1.77, all P>0.05).33 (15.4％) cases developed PE out of 214 pregnant women with PE risk factors .Compared to those who didn′t develop PE , it showed no statistical difference of serum C1q, C3, C4, and factor B levels at 20-26 gestational weeks of the women who subsequently developed PE ( C1q:175 mg/L vs.184 mg/L; C3:1523 mg/L vs.1467 mg/L; C4:230 mg/L vs.229 mg/L;FB:344 mg/L vs.320 mg/L;U=2090.000, 1575.000, 2058.500, 1362.000, all P>0.05).Compared to those of the healthy pregnant controls , it showed no statistical difference of serum C1q, C3 and C4 levels of 20-26 gestational weeks of the women who subsequently developed PE (C1q:175 mg/L vs.190 mg/L; C3:1523 mg/L vs.1428 mg/L; C4:230 mg/L vs.227 mg/L; U=353.000, 395.000, 493.500, all P >0.05),while it showed statistical difference (344 mg/L vs.306 mg/L;U=233.500, P=0.007) for factor B.Conclusions Serum C1q level of PE patients significantly decreased, which can be used as potential indicators of PE diagnosis , but serum C1q, C3, C4 level of 20-26 gestational weeks cannot predict risk of PE .Factor B cannot serve as serum index of PE diagnosis , but its serum levels at 20-26 gestational weeks werer higher than those of normal pregnant controls , factor B may be a potential predictor , but need further verification .

Efficacy of Baofukang Suppositories in the Treatment of Cervical Columnar Epithelial Ectopic and Influence on the Levels of ICMI-1mRNA, TGF-β1m RNA and Inflammatory Cytokines in Cervical Tissues / 中国药师

Objective: To investigate the efficacy of Baofukang suppositories on cervical columnar epithelial ectopic and the influence on the expression of uterine tissue intercellular adhesion molecule 1 (ICMI-1), transforming growth factor b1 (TGF-b1) mRNA and inflammatory cytokines in cervical tissues.Methods: Totally 102 patients with cervical columnar epithelial ectopic were randomly divided into the observation group and the control group with 51 ones in each.The observation group received Baofukang suppositories, and the control group was treated with cryosurgery.The clinical efficacy was compared between the groups, and before and after the treatment, the levels of ICMI-1 mRNA, TGF-b1 mRNA and inflammatory cytokines were also recorded and compared.Results: The clinical curative effect of the observation group was 94.12%,which was better than that of the control group (P＜0.05).After the treatment, the levels of ICMI-1 mRNA and TGF-b1 mRNA decreased when compared with those before the treatment (P 0.05), while those in the observation group decreased after the treatment (P <0.05), which were significantly lower than those in the control group (P＜0.05).There were no significant changes in menstrual period and menstrual cycle in the two groups before and after the treatment, and no adverse reactions occurred during the treatment.Conclusion: Baofukang suppositories are effective in the treatment of columnar epithelial ectopic, which can reduce the levels of ICMI-1 mRNA, TGF-b1 mRNA and serum inflammatory cytokine in the patients.

Construction and assessment of recombinant plasmid pRNAT-U6.1/CFB siRNA and its inhibitory effect on proliferation of human umbilical vein endothelial cells / 中华实验眼科杂志

Background Choroidal neovascularization (CNV) is one of the causes of blindness in multiple eye diseases.Researches showed that complement system participates in the pathogenesis of CNV.Objective This study was to construct the recombinant of complement factor B-small interference RNA (CFB-siRNA) expression vector and to observe its inhibitory effect on human umbilical vein endothelial cells (ECV-304).Methods CFB gene primers were designed based on human CFB gene,and an expression vector of CFB-siRNA was constructed by inserting CFB-siRNA into pRNAT-U6.1/Neo plasmid.Recombinant plasmids were confirmed by the digestion analysis of restriction endonuclease,and all inserted sequences were verified by DNA sequencing.The recombinant pRNAT-U6.1/CFB-siRNA plasmid and the blank plasmid were transfected into ECV-304 cells in the CFB-siRNA group and blank plasmid group by electroblot,respectively,and non-transfected cells served as the normal control group.The cells were observed under the fluorescence microscope 48 hours after transfection,and the transfective efficiency was calculated.The relative expression of CFB mRNA in the cells of different groups was detected by semi-quantitative reverse transcription PCR (RT-PCR).MTT was employed to calculated the growth inhibitory rates of the cells 24,48 and 72 hours after transfection.The percentages of the cells in different cell cycles were detected by flow cytometry.Results The sequence of the target vector was identical to the designed sequence.The green fluorescence protein (GFP) was seen in both the CFB-siRNA group and the blank plasmid group.The relative expression levels of CFB mRNA were 0.07 ±0.04,0.14 ±0.02 and 0.14 ±0.03 in the CFB-siRNA group,the blank plasmid group and the normal control group,respectively,a significant difference was obtained among the three groups (F=233.05,P =0.00);the expression level of CFB mRNA in the CFB-siRNA group was significantly declined in comparison with the blank plasmid group and the normal control group (both at P＜0.05).The growth inhibitory rates of the cells were (23.45 ±0.01) ％,(33.48 ±0.02) ％ and (45.49±0.01) ％ at 24,48 and 72 hours after transfection,respectively,a significant difference was obtained among the three groups (Fgroup =212.99,P =0.00);the growth inhibitory rates in CFB-siRNA group were significantly higher than that in the blank plasmid group and normal control group (all at P＜ 0.05).The percentages of G1 phase cells were (44.4 ±0.5) ％,(25.8 ±0.4) ％ and (27.9 ± 0.6) ％ in the CFB-siRNA group,the blank plasmid group and the normal control group respectively,a significant difference was obtained among the three groups (F=58.98,P=0.00).The percentages of G1 phase and G2 phase cells in the CFB-siRNA group were significantly higher than those in the blank plasmid group and the normal control group (all at P＜0.05).Conclusions Recombinant pRNAT-U6.1/CFB siRNA inhibits the proliferation of ECV-304 cells effectively by arresting the cells in G1 intermediate phase of the growth cycle.

Study on activity of VEGF-binding factors in Paris polyphylla / 中草药

Objective: To screen and isolate vascular endothelial growth factor (VEGF)-binding factors from Paris polyphylla var. yunnanensis, and explore their property for stimulating VEGF activity. Methods: The VEGF-binding factors from water extract of P. polyphylla var. yunnanensis were screened by VEGF-affinity chromatography and further purified by HPLC method. Their activity on proliferation of VEGF-dependent cells was determined by MTT analysis with sensitive cell line HepG2 as model. We also predicted the possible components according to RRLC-QTOF and UV results. Results: The VEGF-binding factors screened from the water extract of P. polyphylla var. yunnanensis were named as factor B3 which included three compounds and could induce the proliferation of VEGF-dependent cell line HepG2 but not the VEGF-independent cell line HEK293. Further studies indicated that factor B3 had an additive effect with VEGF to induce the proliferation of HepG2, and the additive effect could be attenuated by VEGF antibody. In addition, the proliferation of HepG2 cells induced by factor B3 alone could also be attenuated by VEGF antibody. Furthermore, based on the results of RRLC-QTOF and UV analysis, we predicted that factor B3 are probably members of saponin family. Conclusion: The factor B3 isolated from P. polyphylla var. yunnanensis has the property to stimulate VEGF activity.

The role and significance of serum B factor in the pathogenesis of systemic lupus erythematosus / 中国医师杂志

Objective To investigate the expression of serum B factor in the peripheral blood of patients with systemic lupus erythematosus (SLE),and explore its role in the pathogenesis.Methods Seventy eight patients with SLE in our hospital and 46 healthy persons were eligible to participate in this study.Rate nphelometyr was used to test serum B factor for 78 patients with SLE and 46 healthy controls.According to systemic lupus erythematosus disease activity index (SLEDAI),participants were divided into steady SLE group (SLEDAI ＜ 5) and active SLE group (SLEDAI ≥5),which was further divided into mild,moderate,and serious subgroups.The differences in serum B factor between SLE patients and healthy controls,including SLE patients with different severity,were all compared.Then we analyzed the differences in serum B factor and other laboratory and clinical indexes between active and steady SLE patients.The correction of serum B factor and other laboratory and clinical indexes were also analyzed.Results Compared to healthy controls,patients with SLE had significantly lower value of serum B factor [(27.13 ± 8.98) mg/dl vs (36.73 ± 5.47) mg/dl,t =7.4,P ＜ 0.01].Compared to steady SLE group,SLE active group had significantly lower level of serum factor B,C3 and C4,and also had significant higher level of C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) (all P ＜ 0.05).Moreover,There were significant differences in the lower level of serum B factor between subgroups.Correlation analysis showed that the level of serum B factor was negatively associated with the levels of CRP and SLEDAI scores,whereas serum B factor was positively associated with the levels of C3 and C4 (all P ＜ 0.05).Conclusions Serum B factor is related to SLE.Serum B factor might be involved in the pathogenesis of SLE.Detection of serum B factor is helpful for diagnosis and evaluation of SLE disease activity.

The Clumping factor B gene typing in discriminating the strains of nosocomial-acquired methicillin-resistant Staphylococcus aureus / 中华流行病学杂志

Objective To evaluate the clfB typing method in discriminating the ST239 methicillin-resistant Staphylococcus aureus (MRSA) isolated from patients under nosocomial infection in Tianjin first central hospital so as to access the clinical risk factors and outcomes of the MRSA nosocomial infection from ICU and non-ICU departments.Methods Forty-two stains of MRSA with known SCCmec type were chosen in both ICU (n=35) and non-ICU (n=7) wards,from 2006 to 2012,of which MLST genotype was ST239.Clinical risk factors and rates on drug resistant to MRSA were counted,respectively.Results All the isolates of MRSA belonged to the same lineage 3 and 6 heplotypes,based on clfB variable-number random repeats typing.Thirty-five isolates from ICU belonged to 6 heplotypes,among which clfB3-52,3-52E,3-50,3-52C,3-50A and 3-50E were accouted for 42.9％,37.1％,8.6％,5.7％,2.9％ and 2.9％,respectively.Seven isolates from non-ICU belonged to 3 heplotypes,in which 3-52,3-52E and 3-50 were accouted for 42.8％,28.6％,28.6％,respectively.When clfB typing was combined with SCCmec typing in use,results showed that the index of discrimination as 0.767,better than clfB (ID=0.688) or SCCmec (ID=0.303) used alone.SCCmec Ⅲ-clfB3-52E seemed as the major clone among the 10 heplotypes of clfB/SCCmec typing,which was accounted for 40.4％.There were significant differences on the length of hospitalization (P＜0.005) and the duration of antibiotics use (P＜0.05) between ICU and non-ICU.Conclusion The clfB typing method which was based on variable-numbers of tandom repeats showed powerful ability of resolution.It could also be combined with MLST and SCCmec typing to be used in local epidemiological investigations.

Relationship between vascular endothelial growth factor-B and ocular neovascularization disease / 中华实验眼科杂志

Abnormal neovascularization can lead to a variety of eye diseases,and vascular endothelial growth factor(VEGF) plays an important role in the generation and development of neovascularization diseases.As a member of VEGF family,VEGF-B can regulate neovascularization by the effect of vascular survival and cell apoptosis,in order to inhibit the generation and development of neovascularizition,while it has an insignificant impact to angiogenesis and vascular permeability.Meanwhile,VEGF-B can protect cardiac and neuron.As a method of clearly effective to treat neovascularization disease,anti-VEGF attracts extensive attention.Because of the dual character of VEGF-B,we need more studies on the effect of treating ocular neovascularization disease by the drug of anti-VEGF-B.This review summarizes the role of VEGF-B in blood vessel,cell,neuron and cardiac of body and its application in the treatment of ocular neovascularization disease.

DNA immunization of Mycobacterium tuberculosis resuscitation-promoting factor B elicits polyfunctional CD8+ T cell responses

PURPOSE: T cell-mediated immune responses, and particularly activation of polyfunctional T cells that simultaneously produce multiple cytokines, are necessary for the control of Mycobacterium tuberculosis. In the present study, we examined if DNA immunization of Mycobacterium tuberculosis resuscitation-promoting factor B (RpfB) elicits polyfunctional T cell responses in mice. MATERIALS AND METHODS: C57BL/6 mice were immunized intramuscularly three times, at 3-week intervals, with RpfB-expressing plasmid DNA. For comparison, protein immunization was performed with recombinant RpfB in control mice. After immunization, RpfB-specific T cell responses were assessed by interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot assay and intracellular cytokine staining (ICS), and T cell polyfunctionality was assessed from the ICS data. RESULTS: RpfB DNA immunization induced not only humoral immune responses, but also CD8+ and CD4+ T cell responses. Immunodominant T-cell epitopes were identified within RpfB by assays with overlapping peptides. RpfB DNA immunization elicited a polyfunctional CD8+ T cell response that was dominated by a functional phenotype of IFN-gamma+/TNF-alpha+/IL-2-/CD107a+. CONCLUSION: RpfB DNA immunization elicits polyfunctional CD8+ T cell responses, suggesting that RpfB DNA immunization might induce protective immunity against tuberculosis.

Genetic association of complement factor B gene polymorphisms with susceptibility and prognosis of IgA nephropathy / 中华肾脏病杂志

Objective To investigate the genetic association of complement factor B (CFB)gene polymorphisms with the susceptibility and prognosis of IgA nephropathy (IgAN).Methods Four hundreds and sixty-three Northern Han Chinese patients with IgAN and two hundreds and ninty-six geographically and ethnic matched healthy volunteers were recruited.Peripheral blood was collected from recruited individuals for DNA extracting.After amplified by plymerase chain reaction (PCR),genotyping of the four single nucleotide polymorphisms (SNPs) in CFB gene,which were rs4151667,rs12614,rs641153 and rs117314762,were performed by sequencing.Differences of allele and genotype frequencies were analyzed between IgAN patients and healthy controls.Moreover,the association between these SNPs and disease clinical manifestation,pathological features and long term renal outcome in IgAN patients were further analyzed.Results The G allele and GG genotype frequencies of rs117314762 in CFB gene were significantly higher in IgAN patients than that in healthy controls.No difference in allele and genotype frequencies of rs4151667,rs12614,rs641153 between IgAN patients and healthy controls was observed.Furthermore,no association was found between these SNPs in CFB gene and clinical manifestation,pathological features and long term renal outcome of IgAN.Conclusion Association between rs117314762 in CFB gene and IgAN susceptibility suggests that there may be functional variants in CFB gene or its linked genetic region,which needs further exploration.

The low frequency of Y chromosome microdeletions in subfertile males in a Sinhalese population of Sri Lanka.

Aims: This study was designed to determine the prevalence of azoospermia factor (AZF) microdeletions on the Y chromosome in Sri Lankan Sinhalese infertile men with azoospermia and severe oligozoospermia. Settings and Design: The patient group was 207 karyotypically normal infertile Sinhalese males. Materials and Methods: The presence of 13 sequence-tagged site (STS) markers in the AZF region was tested using multiplex polymerase chain reaction (M-PCR). One hundred and twenty unselected men were also studied as a control group. Results: Three (1.5%) had classic Y chromosome microdeletions in the AZFc sub-region. Conclusions: These results suggest a much lower Y chromosome microdeletion frequency than previously thought, even among a strictly selected group of sub-fertile males in Sri Lanka.

Alternative complement pathway and factor B activities in rats with altered blood levels of thyroid hormone

Evaluating the activity of the complement system under conditions of altered thyroid hormone levels might help elucidate the role of complement in triggering autoimmune processes. Here, we investigated alternative pathway (AP) activity in male Wistar rats (180 ± 10 g) after altering their thyroid hormone levels by treatment with triiodothyronine (T3), propylthiouracil (PTU) or thyroidectomy. T3 and thyroxine (T4) levels were determined by chemiluminescence assays. Hemolytic assays were performed to evaluate the lytic activity of the AP. Factor B activity was evaluated using factor B-deficient serum. An anti-human factor B antibody was used to measure factor B levels in serum by radial immunodiffusion. T3 measurements in thyroidectomized animals or animals treated with PTU demonstrated a significant reduction in hormone levels compared to control. The results showed a reduction in AP lytic activity in rats treated with increasing amounts of T3 (1, 10, or 50 µg). Factor B activity was also decreased in the sera of hyperthyroid rats treated with 1 to 50 µg T3. Additionally, treating rats with 25 µg T3 significantly increased factor B levels in their sera (P < 0.01). In contrast, increased factor B concentration and activity (32 percent) were observed in hypothyroid rats. We conclude that alterations in thyroid hormone levels affect the activity of the AP and factor B, which may in turn affect the roles of AP and factor B in antibody production.

Inhibition efficacy of CFB-siRNA on laser-induced choroidal neovascularization in rat / 中华实验眼科杂志

BackgroundChoriodal neovascularization is an important ocular manifestation of angiogenesis in eyes,which derives from the choroid capillaries.Recent studies have found that complement activation is playing a key role in the laser-induced CNV.Because of the key position of CFB in the alternative pathway,bytargeting CFB and blocking the alternative pathway may provide an approach to observe the role of this alternative pathway in the generation of CNV.Objective This study was to investigate the inhibitory effect of reconstructed complement factor B (CFB)-small interfering ribonucleicacid(siRNA)on choroidal neovascularization (CNV)and its mechanism. Methods Experimental CNV was induced by laser photocoagulation in 96 eyes of 48 clean Brown Norway rats.The rats were randomly divided into 4 groups.25,50 and 75 μg B factor siRNA were injected via caudal vein on 1 day,3,5 days after photocoagulation in different dose groups,and normal saline solution was injected at the same way in experimental control group.Other 12 normal rats were used as blank control group.Fundus fluorescein angiography(FFA) was performed on 3,7,14,21,28 days after injection of CFB-siRNA and CNV was scored.The expressions of vascular endothelial growth factor(VEGF) and factor Ⅷ in choroid were detected by immunochemistry.The expressions of CFB-siRNA,VEGF,transforming growth factor β2( TGF-β2 )proteins in choroid were determined using immunochemistry in 7,14,21,28 days,and the expressions of mRNA of CFB-siRNA,VEGF,TGF-β2 were examined by reverse transcription polymerase chain reaction(RT-PCR). ResultsFFA revealed that the CNV rates in various doses of CFB-siRNA groups were significant lower than those of experimental control group in various time points(P＜0.05),and those in 75 μg B factor siRNA were decreased in comparison with 25 μg B factor siRNA (P＜0.05).Immunochemistry showed that the intensities of the VEGF and factor Ⅶ expression in various doses of CFB-siRNA groups were weaker than the blank control group ( P ＜ 0.05 ).Compared with the control group,the expression of CFB reduced in 7 days,and then approached to the level near the control group.Fourteen to twenty-one days after injection of CFB-siRNA,VEGF and TGF-β2 depressions in different doses of CFB-siRNA groups were lower than blank control group( P＜0.05 ).CFB expression in choroid showed the lower levels in CFB-siRNA injection group compared with blank control group in from 7 through 21 days (P＜0.05).RT-PCR displayed the gradual increase of CFB mRNA and curve-like changes of VEGF and TGF-β2 with time prolong. Conclusions Recombinated CFB-siRNA can effectively inhibit laser-induced CNV by down-regulating the expression of VEGF and factor Ⅷ.Alternative pathway of complement plays an important role in the production of CNV.

Role of NF-?B in the mechanism of liver injury following intestinal perforations due to abdominal firearm wound in pigs / 中国现代普通外科进展

Objective:To study the role of NF- ?B in the mechanism of liver injury following intestinal perforations due to abdominal firearm wound. Methods:A total of 42 Chang-Bai piglets were assigned randomly into 7 groups: control group and wounded 1, 2, 4, 8, 12, 24 hours group.The model of intestinal perforations due to abdominal firearm wound was established in wounded groups. Hepatic NF-?B and TNF-? content was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum ALT levels were also determined at the same time. The alterations of hepatic tissue were observed under light microscope. Results: Levels of hepatic NF-?B activity in wounded groups were significantly elevated compared with control group, and two peaks appeared in 1 h group and 8 h group, respectively (P