1.Construction and assessment of recombinant plasmid pRNAT-U6.1/CFB siRNA and its inhibitory effect on proliferation of human umbilical vein endothelial cells
Huan, TONG ; Qingli, SHANG ; Jingxue, MA ; Jian, GAO ; Xin, WANG
Chinese Journal of Experimental Ophthalmology 2015;33(8):686-690
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.
2.Value of computed tomography and magnetic resonance imaging in diagnosis and differential diagnosis of small hepatocellular carcinoma.
Yan MA ; Xue-lin ZHANG ; Xin-yu LI ; Lin ZHANG ; Huan-huan SU ; Chuan-yin ZHAN
Journal of Southern Medical University 2008;28(12):2235-2238
OBJECTIVETo analyze the computed tomography (CT) and magnetic resonance imaging (MRI) findings of small hepatocellular carcinoma to improve the accuracy in the diagnosis.
METHODSThis retrospective analysis involved 41 patients with small hepatocellular carcinoma cases confirmed by pathological examination of the biopsy samples or follow-up. These patients were assessed for CT and MRI findings including lesion size, density or signal intensity, enhancement patterns, and presence of tumor capsules.
RESULTSOn unenhanced CT images, small hepatocellular carcinomas were displayed mainly as low-density masses, and the majority of tumors presented with low signal intensity on T1-weighted unenhanced MR images with increased signal intensity on T2-weighted images in comparison with the surrounding liver parenchyma. Most of tumors showed intense enhancement during the arterial phase (CT in 15 cases and MRI in 13 cases), but some appeared isointense to the liver parenchyma (CT in 4 cases and MRI in 4 cases). In portal and delayed phases, the tumors typically had lower signal intensity than that of the surrounding liver tissues (CT in 25 cases and MRI in 12 cases) with enhancement of the tumor capsules (13 cases).
CONCLUSIONDynamic enhanced scanning can be more informative of the pathology and blood supply of small hepatocellular carcinoma. Early and late arterial phase imaging may help in detecting the small lesions and in making differential diagnosis.
Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; diagnostic imaging ; Diagnosis, Differential ; Female ; Humans ; Image Enhancement ; Liver Neoplasms ; diagnosis ; diagnostic imaging ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed
3.Research progress on influence of intestinal microflora on peripheral immune organs in mice
Huai-Juan XUE ; Long MA ; Huan QIN ; Xin-Sheng YAO
Chinese Journal of Immunology 2018;34(6):953-956
The study found that the presence of intestinal microbiota is not only important for the metabolism of essential nutrients in the body, but also plays a key role in the development of the body′s immune system in recent years. Partial microbiota, through natural selection and co-evolution with the host, forms symbiotic relationships with host microbes that are inseparable from host physiology in mice. Symbiotic flora affects the formation of the body′s immune system by affecting innate and adaptive immunity and the development of various regulatory mechanisms. The destruction of the microbial ecosystem in the intestine can lead to the occurrence of many diseases,especially those related to the immune system. Peripheral immune organs always receive a number of immune cells colonized by antigen stimulation. So,the intestinal flora plays an important role in maintaining the function of immune cells. This article will investigates the effects of mouse-related intestinal flora on peripheral immune organ function.
4.Reparation of deep burn wounds with different kinds of skin flaps in patients with electrical injury.
Yun-Chuan PAN ; Yan-Kun CHEN ; Xin-Chi MA ; Jia-Qin XU ; Si-Huan CHEN
Chinese Journal of Burns 2004;20(3):174-176
OBJECTIVETo sum up the clinical experience in the reparation of burn wounds with different types of skin flaps in patients with severe electrical injury.
METHODSFree skin flap, axile island flap with vascular pedicle and muscular skin flap were employed to repair 64 wounds in 49 cases.
RESULTSSkin flap necrosis happened in 2 cases, congestion and necrosis at the edge of skin flaps in 4 cases, and complete survival of the skin flaps and primary healing of the wounds in the remaining cases.
CONCLUSIONReparation of wounds with proper skin flaps according to the degree of the electrical injury is effective and reliable, which might be beneficial to the prevention of necrobiosis and the infection in deep tissue.
Adolescent ; Adult ; Burns, Electric ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Surgical Flaps
5.Relationship between liver pathology and clinical characters in HBV carriers.
Li-Na MA ; Zhi-Min HE ; Zhen-Huan CAO ; Yong-Hong ZHANG ; Xin-Yue CHEN
Chinese Journal of Hepatology 2010;18(6):433-436
OBJECTIVESTo study the relationship between liver pathology and clinical characters of chronic HBV carriers.
METHODSAnalyze the age, sex, grade of liver inflammation and stage of liver fibrosis among patients with chronic HBV carriers (n = 58) and non-active HBsAg carriers (n = 32), and compare the grade of liver inflammation and stage of liver fibrosis in different groups according to age, ALT levels and with/without HBeAg. The data was processed by using t test or Chi-square test for statistical analysis, respectively.
RESULTS(1) No differences existed in gender composition ratio between chronic HBV carriers and non-active HBsAg carriers. However, the ages of non-active HBsAg carriers group (35.2+/-7.6) were much older than that of the HBV carriers group (24.7+/-4.8) (t = 2.576, P = 0.017). (2) The stage of liver fibrosis in non-active HBsAg carriers group was more aggravated than that of the chronic HBV carriers group (Chi-square = 23.231, P less than 0.01), whereas no significant differences existed between these 2 groups (Chi-square = 0.058, P = 0.972). (3) As to the grade of liver inflammation and the stage of liver fibrosis, significant differences existed between the groups with higher level of serum ALT (20-40 U/L) and lower level ( is less than or equal to 20 U/L) (Chi-square = 7.827, P = 0.008; Chi-square = 14.303, P = 0.001), and similar results also existed between elder group (more than 40) and younger group (is less than or equal to 40) (Chi-square = 10.949, P = 0.001; Chi-square = 21.271, P less than 0.01); (4) Among the chronic HBV carriers, significant differences existed in grade of liver inflammation between groups with HBeAg positive and negative patients (Chi-square = 10.275, P = 0.002), and the latter was more aggravated; however, there was no difference in stage of liver fibrosis between them (Chi-square test = 3.457, P = 0.178).
CONCLUSIONLiver histopathology can be recommended to guide the clinical diagnosis and treatment, especially for the chronic HBV carriers, with elder age, ALT close to normal and HBeAg negative.
Adolescent ; Adult ; Age Factors ; Biopsy ; Carrier State ; pathology ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; Hepatitis B, Chronic ; pathology ; virology ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult
6.Glycated haemoglobin in diagnosis of diabetes mellitus and pre-diabetes among middle-aged and elderly population: Shanghai Changfeng study.
Hui MA ; Xin GAO ; Huan Dong LIN ; Yu HU ; Xiao Ming LI ; Jian GAO ; Nai Qing ZHAO
Biomedical and Environmental Sciences 2013;26(3):155-162
OBJECTIVETo investigate the optimal glycated haemoglobin (HbA1c) cut off points and evaluate the impact of HbA1c on diabetes and pre-diabetes in middle-aged and elderly population.
METHODSSubjects were recruited from Shanghai Changfeng Study. A total of 1973 community-based participants (age ⋝45) without known diabetes underwent oral glucose tolerance test (OGTT) by using a 75-g oral glucose load and HbA1c was measured by using high performance liquid chromatography (HPLC). Subjects were classified as normal glucose tolerance (NGT), pre-diabetes(impaired glucose regulation, IGR) and new diagnosed diabetes (NDD) per 1999 WHO criteria. Two tests are compared with receiver operating characteristic curve (ROC).
RESULTSAmong 1973 subjects, 271 (13.7%) were diagnosed as NDD and 474 (24.0%) as IGR by using OGTT. HbA1c was 5.7%±0.7% in this population. Use of 6.5% as the HbA1C cutoff point has sensitivity of 38.7% and specificity of 98.5%. We recommend 6.0% as a better cutoff value for diagnosis of diabetes in this population (AUC 0.829, 95% CI 0.798-0.860, P<0.001) with its sensitivity and specificity as 66.1% and 86.8%. For IGR, the results showed low sensitivity (44.9%) and specificity (66.7%) with an AUC of 0.571 for HbA1c when 5.8% was used as the cutoff point. Participants detected with HbA1c⋝6.0% were associated with nearly the same metabolic characteristics, including body mass index (BMI), blood pressure, lipid profile and urine albumin-creatinine ratio (uACR) compared with diabetic subjects detected by OGTT.
CONCLUSIONThe optimum HbA1c cutoff point for diabetes in our study population was lower than ADA criteria, and HbA1c may not be used to identify IGR.
Aged ; China ; Diabetes Mellitus ; diagnosis ; metabolism ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Male ; Middle Aged ; Prediabetic State ; diagnosis ; metabolism
7.Acupuncture combined with auricle cutting method for blood stasis-type psoriasis: a randomized controlled trial.
Ting LI ; Zhi-Yan LIU ; Huan YANG ; Zhong MA ; Hong-Yan QU ; Yu LI ; Hai-Bin HUANG ; Juan LIU ; Jie LI ; Ji-Xin WU
Chinese Acupuncture & Moxibustion 2014;34(5):449-453
OBJECTIVETo verify the clinical efficacy of acupuncture combined with auricle cutting method for treatment of blood stasis-type psoriasis.
METHODSFifty-six cases of blood stasis-type psoriasis were randomly divided into a combined therapy group, a auricle cutting group, an acupuncture group and a control group, 14 cases in each one. Based on regular treatment of TCM decoction in four groups, the combined therapy group was treated with acupuncture and auricle cutting method, and the auricle cutting group was treated with sham-acupuncture and auricle cutting, and the acupuncture group was treated with acupuncture and sham auricle cutting, and the control group was treated with sham-acupuncture and sham auricle cutting. The acupuncture was applied at Dazhui (GV 14), Feishu (BL 13), Ganshu (BL 18) and Geshu (BL 17), etc., and manipulated with routine technique; in the sham acupuncture, the needle was inserted into dermis layer so that the needles could be swung without being dropped out. In the auricle cutting, erbeixin (P1) of unilateral auricle was selected and cut by Chan needle to perform bloodletting; in the sham auricle cutting, the neighborhood approximately 0.5 cm next to erbeixin (P) of auricle was selected as cutting area. The treatment was given once a day, seven days as a treatment session for totally two sessions. Psoriasis area and severity index (PASI) before and after treatment was observed and efficacy of each group was compared.
RESULTSThe effective rate was 57.1% (8/14) in the combined therapy group, which was superior to 14.3% (2/14) in the auricle cutting group, 7.1% (1/14) in the acupuncture group and 0.0% (0/14) in the control group (all P < 0.05). The scores of PASI were all decreased in each group after the treatment (all P < 0.05), which was the most significant in the combined therapy group (all P < 0.05). After factorial analysis, the main effect was P < 0.05 in the auricle cutting, P < 0.05 in the acupuncture and P < 0.05 in interaction effect of combined therapy.
CONCLUSIONThe scores of PASI of blood stasis-type psoriasis could be effectively reduced by acupuncture, auricle cutting method and TCM decoction, among which the interaction effect of auricle cutting and acupuncture combined with TCM decoction is the most significant.
Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Bloodletting ; Combined Modality Therapy ; Ear ; blood supply ; Female ; Humans ; Male ; Middle Aged ; Psoriasis ; blood ; therapy ; Treatment Outcome ; Young Adult
8.Clinical analysis of 11 children with primary osteoma cutis
Jiaosheng XU ; Xin XIANG ; Zigang XU ; Huan XING ; Lin MA
Chinese Journal of Dermatology 2019;52(8):525-528
Objective To investigate clinical manifestations,pathological features and outcomes of primary osteoma cutis in children.Methods Eleven children with confirmed primary osteoma cutis diagnosed in Department of Dermatology,Beijing Children's Hospital between 2011 and 2018 were included into this study.The clinical manifestations,histopathological features,and outcomes were analyzed retrospectively.Results Among the 11 patients,7 were males and 4 were females.Primary osteoma cutis occurred within 22 months after birth in all the children,the median age of onset was 1 month,and the disease occurred during the first 6 months of life in 10 children.The skin lesions were characterized by skincolored or reddish indurated papules,plaques or nodules of varying size with slight epidermal atrophy.Three patients had local skin lesions,and 8 had multiple skin lesions.Serum calcium and parathyroid hormone levels were within normal limits in all the children,and no developmental deformity was observed at birth in any of the children.Histopathological examination revealed the formation of mature lamellar bone in the dermis in all the cases,which involved the subcutaneous adipose tissue in 5 cases.The skin lesions became stable 8-18 months after the occurrence in 10 patients,which was consistent with primary plaquelike osteoma cutis.Only 1 patient underwent a slowly progressive course,and the skin lesions involved subcutaneous deep tissues,leading to dyskinesia,which was consistent with progressive osseous heteroplasia.Conclusions Primary osteoma cutis in children mostly occurs in infancy,whose clinical manifestations are atrophic,indurated plaques or nodules,and its main pathological feature is the formation of mature lamellar bone.Long-term follow-up is needed,and attention should be paid to the occurrence of progressive osseous heteroplasia.
9.Etiology and genetic diagnosis of short stature in children.
Wei-Wei CHEN ; Huan-Xin LIU ; Jing LIU ; Lin-Lin YANG ; Min LIU ; Hui-Juan MA
Chinese Journal of Contemporary Pediatrics 2019;21(4):381-386
OBJECTIVE:
To study the etiology and genetic diagnosis of children with short stature.
METHODS:
A retrospective analysis was performed to study the etiological distribution and clinical features of 86 children with short stature.
RESULTS:
A total of 6 causes were observed in these children, among which idiopathic short stature (ISS, 41%) and growth hormone deficiency (GHD, 29%) were the most common causes, followed by genetic diseases (14%). There were no significant differences in age at the time of diagnosis, body height, body length and weight at birth, body height of parents and insulin-like growth factor-1 levels between the genetic disease group and the ISS/GHD groups (P>0.05). Compared with the ISS group, the genetic disease group had significantly lower deviation from the 3rd percentile for the height of children of the same age and sex (ΔP3) and height standard deviation score (P<0.05), while there were no significant differences between the genetic disease and GHD groups (P>0.05). The analysis of the clinical manifestations for the genetic disease group showed heterogeneity and phenotypic overlap in children with different genetic diseases.
CONCLUSIONS
ISS, GHD and genetic diseases are major causes of short stature in children. For children with severe short stature, genetic testing should be performed to make a definitive diagnosis after GHD has been excluded.
Body Height
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Child
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Dwarfism, Pituitary
;
Genetic Testing
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Growth Disorders
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Human Growth Hormone
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Humans
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Retrospective Studies
10.Effects of total flavonoids of Oxytropis falcata Bunge on CCl4-induced liver fibrosis in rats
Tian-Yan YANG ; Xin-Huan MA ; Zhi-Wei XU ; Rong-Kun LI ; Fang-Xiong MA ; Bao-Feng HE ; Liang CHEN ; Xiao-Qing CHEN ; Jun ZHANG
The Chinese Journal of Clinical Pharmacology 2024;40(14):2073-2077
Objective To investigate the effects of total flavones from Oxytropis falcata Bunge on hepatic fibrosis(HF)induced by carbon tetrachloride and liver transforming growth factor(TGF-β)/Smad signaling pathway.Methods Forty-eight male rats were randomly divided into normal group(intraperitoneal injection of peanut oil,intragastric administration of 0.9%NaCl),model group(intraperitoneal injection of 40%CC14 peanut oil solution induced HF model,intragastric administration of 0.9%NaCl),positive control group(modeling,intragastric administration of 0.2 mg·kg-1 of colchicine),experimental-L,-M,-H groups(modeling,intragastric administration of 100,200 and 400 mg·kg-1 of total flavonoid extract of Oxytropis falcata Bunge),8 individuals in each group,for 4 consecutive weeks.The histopathological changes were observed by hematoxylin-eosin and Masson staining.Serum liver function and liver fibrosis were measured;erum inflammatory factors were detected;fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to determine gene expression in liver.Results The pathological injury of liver tissue in the model group was serious,and a large number of inflammatory factors and collagen fibers were accumulated,while the rest of the treatment groups had different degrees of remission.In normal group,model group,positive control group,experimental-L,-M,-H groups,glutamic-pyruvic transaminase levels were(49.28±12.44),(5 885.42±948.37),(4 454.60±489.27),(4 650.47±843.53),(3 761.75±887.30)and(3 544.90±1 066.75)μg·L-1;glutamic-oxaloacetic transaminase levels were(186.90±46.89),(5 936.23±793.81),(3 971.37±780.28),(4 360.30±863.35),(3 943.10±439.47)and(3 971.38±631.08)μg·L-1;hyaluronic acid levels were(45.08±17.16),(104.32±36.06),(66.83±20.09),(70.30±21.07),(60.00±9.68)and(59.02±10.73)μg·L-1;laminin levels were(23.13±3.89),(60.85±13.66),(35.67±9.92),(39.98±9.39),(36.55±12.21)and(34.68±24.83)μg·L-1;type Ⅲ procollagen level were(24.98±5.34),(82.58±30.14),(40.70±16.14),(51.08±23.21),(43.60±12.48)and(44.20±11.66)p±g·L-1;interleukin(IL)-1β levels were(37.63±1.24),(46.10±3.23),(39.22±2.36),(41.33±0.93),(40.25±2.04)and(39.18±2.23)pg·mL-1;tumor necrosis factor-α levels were(314.58±20.56),(383.71±16.97),(349.00±7.93),(348.88±25.11),(325.75±27.84)and(335.07±21.33)pg·mL-1;TGF-β1 mRNA expression of relative quantity respectively were 1.00±0.00,60.99±15.70,9.61±1.59,7.37±1.09,6.41±0.64,6.87±1.09;Smad7 mRNA relative expression were 1.00±0.00,0.34±0.05,0.21±0.03,0.35±0.02,0.38±0.02,0.42±0.03.The above indexes in the model group were compared with the normal group,and the above indexes in the experimental-M,-H groups were compared with the model group,and the differences were statistically significant(P<0.05,P<0.01,P<0.001).Conclusion Total flavonoids of Oxytropis falcata Bunge have protective effects on CC14-induced liver fibrosis in rats,and the mechanism may be related to the regulation of TGF-β/Smad pathway.