1.Construction of CDK7 siRNA expression vector and its effect on cisplatin sensitivity of endometrial carcinoma cells
Wenxin LIU ; Ying CHEN ; Quan HAO
Chinese Journal of Clinical Oncology 2013;(15):893-897
Objective:This study was aimed to investigate the influence of CDK7 siRNA on the sensitivity of endometrial carci-noma cell line HEC-1-A to cisplatin (DDP)-based chemotherapy. Methods:Different CDK7 siRNA fragments were synthesized based on the designs of the CDK gene sequence and were transfected into HEC-1-A. Real time reverse transcription polymerase chain reac-tion (RT-PCR) and Western blot analysis were employed to demonstrate the effects of transfection. The best CDK7 siRNA was chosen to specifically silence CDK7 expression in HEC-1-A.The sensitivity of the cells to DDP therapy before and after transfection was deter-mined by methyl thiazol tetrazolium (MTT) cytotoxicity assay, flow cytometry, and Hoechst/PI double-staining fluorescence microsco-py. Results:A total of four different CDK7 siRNA segments were designed and successfully transfected into HEC-1-A cells. The inter-ference effect in each group was confirmed by real time RT-PCR and Western blot assays. CDK7-423 was determined as the best per-forming CDK7 siRNA (over 70%) to transfect into HEC-1-A cells. MTT cytotoxicity test showed that IC50 of DDP decreased to a range from 45.122 μg/mL and 3.200 μg/mL after inhibition of CDK7 expression. DDP toxicity to the endometrial carcinoma cells sig-nificantly increased (P<0.05). Flow cytometry revealed that the average cell apoptosis rate significantly increased after the inhibition of CDK7 expression (11.66%to 37.57%, P<0.05). Similar results were observed using Hoechst/PI double-staining fluorescence microsco-py, and the number of apoptotic corpuscle demonstrated an apparent increase in the low CDK7-expressing group compared with the pa-rental cells. Conclusion:After the downregulation of CDK7 expression by CDK7 siRNA transfection, DDP chemotherapy sensitivity and apoptosis of endometrial carcinoma cells significantly increased. Further research is anticipated on the use of CDK7 as a new treat-ment target for endometrial carcinoma.
2.Chinese herbal medicine and therapeutic exercise for relieving spasticity in hemiplegic patients
Ying ZHANG ; Lei QIAO ; Hao CHEN
Chinese Journal of Physical Medicine and Rehabilitation 2008;30(6):390-393
Objective To observe the effectiveness of Chinese herbal medicine plus movement therapy in treating upper limb spasticity in hemiplegic patients. Methods Sixty-six patients were divided randomly into two groups. Thirty-five patients in the therapy group were treated with Chinese herbal medicine and movement therapy, and 31 patients in the control group were treated with movement therapy alone. The modified Ashworth scale and sur- face electromyography were applied to evaluate spasticity. Results The total effectiveness rates were 94.3% in the therapy group and 64.5% in the control group. After treatment, the average integrated electromyogram (IEMG) val- ue in the therapy group was lower than that in the control group. The difference in IEMG values before and after treat- ment in the therapy group was significantly larger than in the control group. Conclusion Chinese herbal medicine plus movement therapy is more effective than movement therapy alone in relieving spasticity.
3.Clinical and genetic study of spinocerebellar ataxias 3 within a consanguinity family
Ying HAO ; Yuanyuan CHEN ; Weihong GU ; Guoxiang WANG ; Kang WANG
Chinese Journal of Neurology 2013;(5):329-333
Objective To investigate the inheritance principle of the expanded GAG repeat allele and the clinical features of spinocerebellar ataxias 3 (SCA3) in a consanguinity family with first cousin marriage.Methods The CAG repeats of SCA3 gene were amplified by means of polymerase chain reaction.Fragment analysis with laser-induced fluorescence in capillary electrophoresis were performed for the positive samples detected by agarose gel electrophoresis.Furthermore,the clinical features were analyzed carefully.Results Fragment analysis revealed that the proband carried 2 alleles with 56 and 72 CAG repeats separately.The proband' s father carried 28 and 66,and the expanded CAG repeat allele inherited from his grandfather.The proband' s mother carried 33 and 56,and the expanded CAG repeat allele inherited from his grandmother.The proband' s son carried 27 and 85 and presented with dystonia besides ataxia.Conclusions The proband' s parents have the common ancestors.Their alleles with expanded CAG repeats probably come from the same allele of their ancestor.The GAG repeat is more unstable in the paternal inheritance than in the maternal inheritance.The 71-year-old asymptomatic family member carry the allele with 56 CAG repeats,which indicates the 56 CAG repeats may be not associated with the disease.The patients within this family have variable clinical features,especially the juvenile-onset case presents with apparent dystonia.
4.Risk factors for sexual dysfunction in male patients with obstructive sleep apnea and hypoventilation syndrome
Song LIU ; Ying CHEN ; Gangyue HAO ; Zhengyi HE
Clinical Medicine of China 2013;(1):27-30
Objective To investigate the risk factors of sexual dysfunction in male patients with obstructive sleep apnea and hypoventilation syndrome (OSAHS).Methods One hundred and twenty-one OSAHS male patients aged 22-50 years were stratified into two groups based on scores on the rating scales of international index of erectile function-5 (IIEF-5).Patients' characteristics and polysomnography data of the sexual dysfunction group (n =56) were compared with that of the normal group (n =65).Continuous positive airway pressure(CPAP) treatment was carried out in some patients with abnormal sexual function.Results There were significant differences between the two groups in body mass index (BMI),history of hypertensive disease and diabetes mellitus,the rise of morning blood pressure,respiratory disturbance index (RDI),the longest apnea time(Tmax),the lowest oxygen saturation(LSpO2),percentage of recording time with oxygen saturation of 90% or less(T90%),slow wave sleep(S1 + S2) over total sleep time(TST),rapid wave sleep (S3 + S4) time over TST,and rapid eye movement (REM) sleep time over TST in bivariate analysis (P < 0.05).There were statistical significance of the history of diabetes mellitus,slow wave sleep (S1 + S2) over TST,RDI,and LSpO2shown in Logistics regression analysis,and ORs were 2.10 (1.17-5.01),3.52 (1.33-5.97),1.45 (1.08-2.95),and 0.86 (0.67-0.93) respectively (P < 0.05).Moreover,20 OSAHS patients with sexual dysfunction were revaluated after the CPAP treatment in two months,and 14 patients returned to the normal scores.Conclusion The history of diabetes mellitus,slow wave sleep (S1 + S2) over TST,RDI,and LSpO2 were independent risk factors for sexual dysfunction in male patients with OSAHS,and treatment with CPAP could significantly improve sexual function in male patients.
5.Homozygosity mapping and mutation analysis of a consanguineous marriage family with autosomal recessive cerebellar ataxia
Ying HAO ; Weihong GU ; Yuanyuan CHEN ; Jin ZHANG
Chinese Journal of Neurology 2015;48(5):400-405
Objective To identify the pathogenic gene for a Chinese Han consanguineous marriage family with autosomal recessive cerebellar ataxia by homozygosity mapping and mutation analysis.Methods Six members of the family were enrolled in this study,including 3 patients,the unaffected sibling and their parents of first cousin marriage.After excluding GAA repeats mutation of FXN gene,whole-genome single nucleotide polymorphism (SNP) microarray scanning and homozygosity mapping were performed to localize the candidate gene.The coding regions and intronic flanking sequences of the candidate genes were analyzed.Results Four candidate regions were identified,including 2p25.3,9q22.2-34.3,13q12.3-14.3 and 17p13.The SETX gene localizing in 9q22.2-34.3 that is responsible for ataxia with oculomotor apraxia 2 was analyzed at first.There were 4 mutations in exon 10,including three missense mutations (c.3576T > G,p.D1192E ; c.3754G > A,p.G1252R; c.4156A > G,p.I1386V) and a deletion mutation (c.5084_5087delAGTC,p.Q1695_S1696del).Three patients were homozygous of the 4 mutations,an unaffected sibling was normal,and their parents were heterozygous of 4 mutations.Conclusions The pathogenic haplotype comprising four mutations of the SETX gene was identified in the consanguinity family.c.5084_5087delAGTC (p.Q1695_S1696del) is a novel mutation.The affected individuals of this family were characterized by mild phenotype and slow progress without oculomotor apraxia,indicating the clinical variability of the disease.
6.Chronic pharyngitis treated by garlic moxibustion.
Re-Hua CHEN ; Hao LIN ; Ri-Ying JIN
Chinese Acupuncture & Moxibustion 2012;32(1):58-58
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7.Melanin Synthesis was Affected by Extracts of 22 Kinds Chinese Herbs of Acid Taste: an Experi- mental Study.
Li-ping ZHANG ; Juan LIANG ; Bin CHEN ; Ying-hao WANG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(5):618-622
OBJECTIVETo confirm the inhibitory effect of Chinese herbs of acid taste on melanin synthesis.
METHODSActive ingredients of 22 kinds Chinese herbs of acid tastes were extracted by alkali extraction and acid precipitation, alcohol extraction, and water extraction, respectively, which was then dispensed into 25.00, 12.50, and 6.25 g/L suspension. Their effects on activities of tyrosinase were detected using mushroom-tyrosinase-DOPA speed oxidation. Their inhibition rates on activities of tyrosinase were respectively compared with inhibition rates of 1.0, 0.5, and 0.1 mmol/L arbutin.
RESULTSThe 22 kinds Chinese herbs of acid taste included Cornus Officinalis, Crataegus pinnatifida, dark plum fruit, Schisandra Chinensis, Chaenomeles sinensis Koehne, Reynoutria japonica Houtt, Achyranthes Bidentata, Sanguisorba officinalis L., Semen Ziziphi Spinosae, Herba Ecliptae, blueberry, immature bitter orange, submature bitter orange, Prunus mume Var, Hovenia acerba Lindl., Fructus Mori, Pomegranate Rind, white paeony root, Rosa laevigata Michx., Portulaca oleracea L, Terminalia chebula Retz, Rhus chinensis Mill. Their alkaline extractions showed inhibition to activities of tyrosinase to different degrees except Herba Ecliptae. Of them, the highest inhibition rate (88.49% ± 9.98%) was got by dark plum fruit at 25 g/L, while the lowest inhibition rate (11.22% ± 3.36%) was got by immature bitter orange at 6.25 g/L. Their alcohol extractions showed inhibition to activities of tyrosinase to different degrees except Herba Ecliptae. Of them, the highest inhibition rate (75.92% ± 5.57%) was got by Hovenia acerba Lindl. at 25 g/L, while the lowest inhibition rate (9.60% ± 1.15%) was got by submature bitter orange at 6.25 g/L. Their water extractions all had inhibition on activities of tyrosinase. Of them, the highest inhibition rate (54.23% ± 3.56%) was got by Fructus Mori at 25 g/L, while the lowest inhibition rate (10.25% ± 1.83%) was got by Semen Ziziphi Spinosae at 6.25 g/L. Compared with 1 mmol/L arbutin water solution, alkaline extractions of dark plum fruit, Schisandra Chinensis, Rhus chinensis Mill., Rosa laevigata Michx., blueberry, Chaenomeles sinensis Koehne, Portulaca oleracea L, Fructus Mori, Achyranthes Bidentata, Pomegranate Rind; alcohol extractions of dark plum fruit, Rhus chinensis Mill., Pomegranate Rind, Hovenia acerba Lindl., Crataegus pinnatifida, Achyranthes Bidentata; water extractions of Chaenomeles sinensis Koehne, blueberry, and Fructus Mori at 25 g/L got obviously higher inhibition rates (P < 0.05, P < 0.01). Compared with 0.5 mmol/L arbutin water solution, alcohol extraction of Chaenomeles sinensis Koehne and alcohol extraction of dark plum fruit at 12.5 g/L got obviously higher inhibition rates (P < 0.05, P < 0.01).
CONCLUSIONChinese herbs of acid taste could inhibit melanin synthesis, and its mechanism was related to inhibiting activities of tyrosinase.
Drugs, Chinese Herbal ; pharmacology ; Humans ; Melanins ; metabolism ; Monophenol Monooxygenase ; metabolism ; Oxidation-Reduction ; Schisandra ; Taste
8.Protective Effect of Total Glucosides of Mudan Cortex on Acute Myocardial Ischemia in Mice
Xiaoyan XU ; Ying LI ; Wei WANG ; Hao WANG ; Wei CHEN
Herald of Medicine 2015;(9):1158-1161
Objective To study the protective effect of total glucosides of Mudan cortex(TGM) on acute myocardial ischemia in mice and its mechanism. Methods The total of 60 mice were randomly divided into 5 groups (n = 12),normal control and model control (were given equal capacity of 0.9% sodium chloride solution),and TGM at low,middle,high dose (were given with 50,100,200 mg?kg-1 TGM).The mice were administered once daily for consecutive seven days.After the last administration,the mice in the model control and drug groups were treated by intraperitoneal injection of 15 mg ? kg-1 isoproterenol to to make myocardial ischemia animal model. TGM on the T wave and J point on ECG, and serum lactate dehydrogenase (LDH),myocardial tissue superoxide dismutase (SOD),malondialdehyde (MDA) changes was detected,and the extent of myocardial ischemic injury in mice was measured by Nagar-Olsen staining. Results TGM significantly reduced the displacement of ECG T wave and J point,and improved the related biochemical indexes in mice with myocardial ischemia.The activity of LDH [(898.992± 285.108) μmol?mg-1 ] ,the content of MDA [(11.737 ±5.162) nmol?mg-1 ]in mice treated with TGM at high dose obviously decreased in comparison to the model controls,and the activity of myocardial SOD [(45. 505 ± 20.711) U?mg-1 ] significantly elevated compared with the model control.It was showed that TGM significantly diminished the areas of cardiac muscles ischemia injured via Nagar-Olsen staining. Conclusion TGM has the remarkable protective effect on acute myocardial ischemia injury in mice.
9.Natural science-social science multidisciplinary research in medical research and its research organization
Hao CHEN ; Mei WANG ; Ying LI ; Laixin ZHOU
Chinese Journal of Medical Science Research Management 2016;29(5):375-379
Medical research has entered the era of mega-science.The multidisciplinary collaboration has attracted great attention.besides studies on genes and the environment interaction,multidisciplinary research between natural sciences has received widespread recognition.However muhidisciplinary researches on the interaction between the gene and social factors or behavioral factors on health science deserve more emphasis.This article described development of multidisciplinary research and its features;analyzed importants of conducting multidisciplinary research between natural and social disciplines,and finally provided suggestions on research organization reform in order to promote the multidisciplinary research.
10.IL-10 inhibits cardiac fibroblasts proliferation and phenotype transformation induced by TGF-β1 in rats
Yanjie HAO ; Ying CHEN ; Lin XUE ; Xiaoning HAN ; Wenhui DING
Basic & Clinical Medicine 2015;(9):1182-1187
Objective To examine the effects of IL-10 on cardiac fibroblasts ( CFBs) proliferation and phenotype transformation to myofibroblasts (MyoFbs) induced by transforming growth factor-β1 (TGF-β1);and to investigate the regulating pathways .Methods Cardiac fibroblasts were isolated from cardiac ventricles of neonatal SD rats . The passage 2~4 were used and divided into the following groups for treatment:1) control group, 2) IL-10 reac-tion group, 3) TGF-β1 reaction group, and 4) IL-10 plus TGF-β1 reaction group (TGF-β1 treatment followed with IL-10 pretreatment ) .Cells proliferation was assessed by MTT assay and immunocytochemistry staining for prolifera-ting cell nuclear antigen (PCNA);the phenotype transformation into MyoFbs was assessed by immunocytochemistry of α-smooth muscle actin (α-SMA);extracellular signal related kinase ( ERK1/2) and P38 kinase pathways were assessed by western-blot.Results TGF-β1 (10 μg/L) treatment boosted the proliferation and the expression ofα-SMA significantly (P<0.01), while IL-10 (10, 50 or 100 μg/L) plus TGF-β1 co-treatment induced lower cell proliferation and expression of α-SMA than treating with TGF-β1 alone ( P<0.05 ) , with the inhibitory effect of IL-10 being concentration dependent .TGF-β1 could significantly stimulate the ERK 1/2 and P38 kinase phospho-rylation ( P<0.01 ) , however IL-10 (100 μg/L) plus TGF-β1 co-treatment failed to down-regulated the phospho-rylation of ERK1/2 and P38 kinase compared with TGF-β1 alone ( ERK1/2:P<0.05;P38:P<0.01 ) .Conclu-sions IL-10 can attenuate TGF-β1-induced CFBs proliferation and phenotype transformation to MyoFbs .The in-hibitory effects may explained by a mechanism of inhibiting the activation of ERK 1/2 and P38 kinase .