1.Effects of SP600125 at various concentrations on proliferation and osteogenesis of human adipose-derived stem cells in vitro
Lin WANG ; Huijie GU ; Xiao CHEN ; Yun ZHANG ; Liehu CAO ; Weizong WENG ; Panpan PAN ; Fang JI ; Lei CUI ; Jiacan SU
Chinese Journal of Orthopaedic Trauma 2017;19(2):157-163
Objective To elucidate the effects of SP600125 at different concentrations on the proliferation and osteo-differentiation of human adipose-derived stem cells (hASCs).Methods The hASCs harvested were cocuhured with SP600125 at concentrations of 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmol/L in growth medium (OM group) and in osteogenesis medium (OM group),respectively.The DNA quantitative assay was carried out to evaluate proliferation of the hASCs;flow cytometry was used to determine the effect of SP600125 on the cell cycles of hASCs;Alkaline phosphatase level (ALP) and calcium deposition tests were conducted to observe the effects of SP600125 at different concentrations on osteogenic differentiation of the hASCs.Results The proliferation of hASCs was inhibited by 42.1% when the cells were cocultured with SP600125 at the concentration of 10 μmol/L;the suppression decreased with decreased concentration of SP600125.The hASCs of phase G0/G1 in GM cocultured with SP600125 at the concentration of 10 μmol/L were more than those in GM cocultured with dimethylsulfoxide at the same concentration.ALP test revealed that after 10 days of culture in vitro the staining was more and more weakened and scattered and the ALP activity was more and more decreased with the increased concentration of SP600125.The extracellular calcium deposition of hASCs after 14 days of culture in vitro showed that the size and number of calcium nodules decreased with the increased concentration of SP600125.Conclusion SP600125 can suppress the proliferation and osteogenic differentiation of hASCs in vitro.
2.Proliferation inhibition of human lung adenocarcinoma cell line A549 transfected by RASSF1A gene.
Zheng-hao DENG ; Jian-hua ZHOU ; Hui-qiu CAO ; Ming SHENG ; Ji-fang WENG
Journal of Central South University(Medical Sciences) 2005;30(2):193-196
OBJECTIVE:
To determine the effects of exogenous RASSF1A gene on the proliferation and expression of P65 and subunit of NF-kappaB, in lung adenocarcinoma cell line A549.
METHODS:
pcDNA3.0-RASSF1A and pcDNA3. 0 were introduced into A549 cell line by lipofectin transfection, and the A549 cells stably expressing RASSF1A gene were established by G418 selection. The expression of RASSF1A was detected by Western blotting. The cytobiologic characterizations of the positive clone were analyzed by methythiazoletertraolium (MTT) assay and cytometry. The expressing of P65 was analyzed by RT-PCR and Western blotting.
RESULTS:
A549 cells stably expressing RASSF1A protein were established by lipofection mediated transfection and selected for further study. Compared with the nontransfected and vector transfected cells, the positive clone cells grew more slowly. Flow cytometric data showed that more positive clone cells went into phase G0/G1 and fewer cells went into phase S. The expression of P65 in nuclear protein in positive clone cells was lower than that of the control group while there was no obvious difference between the expression of p65 mRNA and P65 protein in total protein among the 3 groups.
CONCLUSION
RASSF1A gene might suppress the proliferation of A549 cells through blocking the activity of P65 protein.
Adenocarcinoma
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genetics
;
pathology
;
Cell Line, Tumor
;
Cell Proliferation
;
Humans
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Lung Neoplasms
;
genetics
;
pathology
;
RNA, Messenger
;
biosynthesis
;
genetics
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Transcription Factor RelA
;
biosynthesis
;
genetics
;
Transfection
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Tumor Suppressor Proteins
;
biosynthesis
;
genetics
3.Multicenter clinical study on the efficacy and safety of inhalable insulin aerosol in the treatment of type 2 diabetes.
Zhi-hong LIAO ; Ying-li CHEN ; Fang-ping LI ; Xiang YAN ; Hai LU ; Li YAN ; Zhi-guang ZHOU ; Da-long ZHU ; Li-nong JI ; Jian-ping WENG ; null
Chinese Medical Journal 2008;121(13):1159-1164
BACKGROUNDA new inhalable insulin aerosol (Inh-Ins) was developed in China. The aim of this multicenter clinical study was to evaluate the efficacy and safety of this new Inh-Ins as a treatment of type 2 diabetes. Regular porcine insulin (RI) was used as a control.
METHODSThis study is a prospective, randomized, open-label, parallel-group multicenter clinical trial in which 253 qualified patients with type 2 diabetes received the insulin Glargine daily at bedtime plus either a pre-meal Inh-Ins or a pre-meal subcutaneous RI for 12 weeks. HbA1c, fasting plasma glucose (FPG), the 1-hour-postprandial blood glucose (1hPBG) and the 2-hour-postprandial blood glucose (2hPBG) were measured. Events were monitored for adverse effects.
RESULTSAfter 12 weeks, the HbA1c decreased significantly from baseline in both treatment groups, with no significant difference between the two regimens. In the Inh-Ins group, FPG, both 1hPBG and 2hPBG significantly declined from baseline after the 8th- and 12th-weeks of treatment. The reduced values of FPG or 1hPBG between the two groups showed a more significant hypoglycemic effect with the Inh-Ins than the RI. After 12 weeks, the pulmonary carbon monoxide diffusing capacity (DLco) was significantly lower in Inh-Ins group than in the RI. The main side effects of Inh-Ins were coughing, excessive sputum, and hypoglycemia.
CONCLUSIONSInh-Ins was effective in decreasing HbA1c like the RI. It was better in lowering the FPG and the 1hPBG than the RI. Its main side effects were coughing, excessive sputum, and hypoglycemia. Also, Inh-Ins slightly impaired DLco.
Adolescent ; Adult ; Aerosols ; Aged ; Blood Glucose ; analysis ; Body Weight ; drug effects ; Cough ; chemically induced ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Female ; Glycated Hemoglobin A ; analysis ; Humans ; Hypoglycemia ; chemically induced ; Insulin ; administration & dosage ; adverse effects ; Male ; Middle Aged ; Prospective Studies
4.Liver histopathological features influencing HBeAg seroconversion in patients with HBeAg-positive chronic hepatitis B responding to Peg-IFN treatment.
Hua-dong YAN ; Fan-rong JIANG ; Cheng-liang ZHU ; Guo-sheng GAO ; Peng-jian WENG ; Ai-rong HU ; Chang-feng XU ; Yao-ren HU ; Ji-fang SHENG
Chinese Journal of Hepatology 2013;21(5):340-344
OBJECTIVETo investigate the therapeutic efficiency of antiviral treatment with pegylated-interferon (Peg-IFN) for hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) and to explore whether liver histopathological features or other factors influence the HBeAg seroconversion treatment response.
METHODSEighty HBeAg-positive CHB patients with diagnosis confirmed by liver puncture were treated with Peg-IFN(2a or 2b)body weight dose, once weekly). At treatment week 48, the rate of HBeAg seroconversion was determined and used to analyze the influence of liver histopathological features (liver biopsy assessment of: inflammation, graded G0 to G4; fibrosis stage, graded S0 to S4), sex, age, differential levels (pre-treatment baseline vs. week 48 post-treatment) of serum alanine transferase (ALT), and HBV DNA, by binary logistic analysis.
RESULTSAt week 48, the overall rate of HBeAg seroconversion was 30.0%. The rate of HBeAg seroconversion gradually advanced with increased liver inflammation (X2 = 8.435, P = 0.015): 9.09% of the 22 patients with G1; 31.58% of the 38 patients with G2; 47.30% of the 19 patients with G3; the one patient with G4. In contrast, the rate of HBeAg seroconversion showed a much weaker association with liver fibrosis (X2 = 5.917, P = 0.116). Only baseline HBeAg level, and no other baseline index, was significantly different between the patients who achieved HBeAg seroconversion and those who did not. Liver inflammation and baseline HBeAg level were identified as influencing factors of HbeAg seroconversion in response to Peg-IFN treatment.
CONCLUSIONPeg-IFN therapy induces a higher rate of HBeAg seroconversion in HBeAg-positive CHB patients with severe liver inflammation; histological analysis of pre-treatment liver biopsies may help to identify patients most likely to benefit from the antiviral regimen.
Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; pathology ; Humans ; Interferon-alpha ; therapeutic use ; Liver ; pathology ; Male ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Serologic Tests
5.Clinical management of abdominal trauma.
Guo-en FANG ; Tian-hang LUO ; Cheng-hui DU ; Jian-wei BI ; Xu-chao XUE ; Guo WEI ; Zhao-zhang WENG ; Li-ye MA ; Ji-de HUA
Chinese Journal of Traumatology 2008;11(4):239-242
OBJECTIVETo improve the prognosis of patients with abdominal trauma.
METHODSBetween January 1993 and December 2005, 415 patients were enrolled in this research. The patients consisted of 347 males and 68 females with mean age of 36 years (ranging from 3-82 years). All abdominal traumas consisted of closed traumas (360 cases, 86.7%) and open traumas (55 cases, 13.3%).
RESULTSA total of 407 cases (98.1%) were fully recovered from trauma and the other 8 cases (1.9%) died of multiple injuries. The mean injury severity score (ISS) of all patients was 22 while the mean ISS of the patients who died in hospital was 42. Postoperative complications were seen in 9 patients such as infection of incisional wounds (6 cases), pancreatic fistula (2 cases) and intestinal fistula (1 case). All these postoperative complications were cured by the conservative treatment.
CONCLUSIONCareful case history inquisition and physical examination are the basic methods to diagnose abdominal trauma. Focused abdominal ultrasonography is always the initial imaging examination because it is non-invasive and can be performed repeatedly with high accuracy. The doctors should consider the severity of local injuries and the general status of patients during the assessment of abdominal trauma. The principle of treatment is to save lives at first, then to cure the injuries. Unnecessary laparotomy should be avoided to reduce additional surgical trauma.
Abdominal Injuries ; diagnosis ; therapy ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged
6.The experimental study of suppressing silicosis fibrosis.
Ze-ping WENG ; Ji-jun ZHANG ; Wei-wei LIU ; Juan CHEN ; Yi-min LIU ; Wei YU ; Li-juan TANG ; Jia-yu CHEN ; Mao FANG ; Cheng ZHANG ; Geng-xin YE ; Ling-zhen CHEN ; Xue-yun ZHONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(10):740-745
OBJECTIVETo compare the difference of effects on SiO(2)-induced alveolitis and early fibrosis between bone marrow-derived mesenchymal-like stem cells (BM-MSCs) and BM-MSCs transfected by pcDNA3.1-HGF and to explore the mechanism of this effects.
METHODSThe Primary BM-MSCs from Wistar male young rats were cultured and labeled by 4, 6-diamidino-2-phenylindole (DAPI). Fifty Wistar rats were randomly divided into 3 groups:model group (10 rats),which was administered with SiO(2) by the trache, the next day,injected PBS via the tail vein; BM-MSCs group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs via the tail vein; pcDNA3.1-HGF plus BM-MSC group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs transfected by pcDNA3.1-HGF via the tail vein. On the 14th and 28th days after treatment, half of the animals were sacrificed, respectively, and the lungs were harvested for frozen section to observe the cell marked by DAPI. HE staining under a fluorescent microscope, and to observe the pulmonary alveolitis and fibrosis by HE and Masson staining under a light microscope. Western blot assay was used to detect the expression of HGF in rat lungs. The expression levels of tumor necrosis factor-α (TNF-α) in pulmonary tissues were analyzed quantitatively by ELISA. The contents of HYP in pulmonary tissues were analyzed quantitatively by sample hydrolysis method.
RESULTSOn the 14th and 28th days after treatment, the scores of pulmonary alveolitis and early fibrosis in pcDNA3.1-HGF plus BM-MSCs group were 2.36 ± 0.17, 2.8 ± 0.14 and 0.1 ± 0.11, 1.16 ± 0.13, which were significantly lower than those (1.68 ± 0.17, 1.58 ± 0.31 and 0.54 ± 0.15, 1.36 ± 0.13) in BM-MSCs group, also which were significantly lower those (2.36 ± 0.17, 2.80 ± 0.14 and 0.64 ± 0.09, 1.84 ± 0.17) in model group (P < 0.05); On the 14th and 28th days after treatment, the TNF-α contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 280.4 ± 23.11 and 249.78 ± 22.33 pg/mg, which were significantly lower than those (341.58 ± 35.34, 442.29 ± 36.76 pg/mg and 319.51 ± 17.84, 348.53 ± 33.95 pg/mg) in BM-MSCs and model groups (P < 0.05); On the 14th and 28th days after treatment, the HYP contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 0.46 ± 0.04 and 0.65 ± 0.05 µg/mg, which were significantly lower than those (0.63 ± 0.04, 1.04 ± 0.07 µg/mg and 0.72 ± 0.60, 1.39 ± 0.60 µg/mg) in BM-MSCs and model groups (P < 0.05).
CONCLUSIONThe effects of BM-MSCs transfected by pcDNA3.1-HGF on suppressing pulmonary alveolitis and early fibrosis induced by SiO2 were better than those of BM-MSCs. The mechanism may be associated with the reduced pulmonary inflammation.
Animals ; Bone Marrow Cells ; cytology ; Hepatocyte Growth Factor ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; metabolism ; Pulmonary Fibrosis ; chemically induced ; prevention & control ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Silicosis ; prevention & control ; Transfection
7.Bibenzyls and Phenanthrenes from Arundina Graminifolia
Rong HUANG ; Yong-Sheng TAO ; Liang ZHANG ; Shuo-Tong HUANG ; Fang-Ning LOU ; Rui-Xuan WENG ; Ji-Yun YE ; Xiao-Ling WEN ; Yu-Peng LI
Journal of Kunming Medical University 2017;38(11):1-4
Objective To study the bibenzyls and phenanthrenes from Arundina graminifolia.Methods The compounds were extracted by 95% alcohol and isolated by column chromatography on silica gel and Sephadex LH-20.Their structures were identified by spectroscopic analysis (1H NMR and13CNMR).Results Eleven compouds were obtained and identified as batatasin Ⅲ (1),arundinanin (2),2,8-dihydroxy-4,7-dimethoxy-9,10-dihydrophenanthrene (3),shancidin (4),arundinan (5),isoshancidin (6),erianthridin (7),lusianthridin (8),eulophiol (9),flavanthrin (10),orchinol (11).Conclusion Compounds 3,7,9 were isolated from this plant for the first time.
8.Analysis of related risk factors for the refracture of adjacent vertebral body after percutaneous kyphoplasty.
Shen-Yun FANG ; Ji-Kang MIN ; Zhong-You ZENG ; Qiang-Hua ZHANG ; Tao ZHU ; Wei WENG ; Wei-Li ZHANG ; Fang-Ying SHEN ; Ying XIA ; Li-Ying DING
China Journal of Orthopaedics and Traumatology 2021;34(8):705-709
OBJECTIVE:
To investigate the risk factors of vertebral refracture after percutaneous kyphoplasty (PKP) for osteoprotic vertebral compression fractures (OVCFs), and to provide reference for clinical prevention.
METHODS:
A retrospective analysis of 228 OVCFs patients who met the inclusion criteria admitted from November 6, 2013 to December 14, 2018. There were 35 males and 193 females, with a male-to-female ratio of 3∶20, and aged 58 to 91 years with an average of (69.70±7.03) years. All patients were treated with PKP and had complete clinical data. According to whether refracture occurred after operation, they were divided into refracture group (24 cases) and non refracture group (204 cases). Factors that may be related to refracture (including gender, age, surgical segment, number of vertebral bodies in the surgical segment, whether combined with degenerative scoliosis, whether anti-osteoporosis treatment) were included in the univariate analyses, and the single factor analysis of statistically significant risk factors was carried out with multiple Logistic regression analysis to further clarify the independent risk factors for vertebral body refracture after PKP. Survival analysis was performed using the time of vertebral refracture after PKP as the end time of follow up, the occurrence of refracture after PKP as the endpoint event, and the presence or absence of degenerative lateral curvature as a variable factor.
RESULTS:
All 228 patients were followed up for 1.8 to 63.6 months with an average of (28.8±15.6) months, and the refracture rate was 10.5%(24/228). There were statistically significant differences between two groups in age, number of operative vertebral bodies, whether combinedwith degenerative scoliosis and whether anti osteoporosis treatment (
CONCLUSION
Combined scoliosis is an independent risk factor for refracture after OVCFs vertebroplasty, and it is also a possible high-risk factor for refracture after surgery.
Female
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Fractures, Compression/surgery*
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Humans
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Kyphoplasty/adverse effects*
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Male
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Osteoporotic Fractures
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Retrospective Studies
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Risk Factors
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Spinal Fractures/surgery*
;
Vertebral Body
9.Biliverdin Protects the Isolated Rat Lungs from Ischemia-reperfusion Injury via Antioxidative, Anti-inflammatory and Anti-apoptotic Effects.
Wen-Fang TIAN ; Ping WENG ; Qiong SHENG ; Jun-Liang CHEN ; Peng ZHANG ; Ji-Ru ZHANG ; Bin DU ; Min-Chen WU ; Qing-Feng PANG ; Jian-Jun CHU
Chinese Medical Journal 2017;130(7):859-865
BACKGROUNDBiliverdin (BV) has a protective role against ischemia-reperfusion injury (IRI). However, the protective role and potential mechanisms of BV on lung IRI (LIRI) remain to be elucidated. Thus, we aimed to investigate the protective role and potential mechanisms of BV on LIRI.
METHODSLungs were isolated from Sprague-Dawley rats to establish an ex vivo LIRI model. After an initial 15 min stabilization period, the isolated lungs were subjected to ischemia for 60 min, followed by 90 min of reperfusion with or without BV treatment.
RESULTSLungs in the I/R group exhibited significant decrease in tidal volume (1.44 ± 0.23 ml/min in I/R group vs. 2.41 ± 0.31 ml/min in sham group; P< 0.001), lung compliance (0.27 ± 0.06 ml/cmH2O in I/R group vs. 0.44 ± 0.09 ml/cmH2O in sham group; P< 0.001; 1 cmH2O=0.098 kPa), and oxygen partial pressure (PaO2) levels (64.12 ± 12 mmHg in I/R group vs. 114 ± 8.0 mmHg in sham group; P< 0.001; 1 mmHg = 0.133 kPa). In contrast, these parameters in the BV group (2.27 ± 0.37 ml/min of tidal volume, 0.41 ± 0.10 ml/cmH2O of compliance, and 98.7 ± 9.7 mmHg of PaO2) were significantly higher compared with the I/R group (P = 0.004, P< 0.001, and P< 0.001, respectively). Compared to the I/R group, the contents of superoxide dismutase were significantly higher (47.07 ± 7.91 U/mg protein vs. 33.84 ± 10.15 U/mg protein; P = 0.005) while the wet/dry weight ratio (P < 0.01), methane dicarboxylic aldehyde (1.92 ± 0.25 nmol/mg protein vs. 2.67 ± 0.46 nmol/mg protein; P< 0.001), and adenosine triphosphate contents (297.05 ± 47.45 nmol/mg protein vs. 208.09 ± 29.11 nmol/mg protein; P = 0.005) were markedly lower in BV-treated lungs. Histological analysis revealed that BV alleviated LIRI. Furthermore, the expression of inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor-β) was downregulated and the expression of cyclooxygenase-2, inducible nitric oxide synthase, and Jun N-terminal kinase was significantly reduced in BV group (all P< 0.01 compared to I/R group). Finally, the apoptosis index in the BV group was significantly decreased (P < 0.01 compared to I/R group).
CONCLUSIONBV protects lung IRI through its antioxidative, anti-inflammatory, and anti-apoptotic effects.