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1.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-976255

RESUMO

ObjectiveTo explore the relapse status based on the positive and negative syndrome scale (PANSS Scale) and related factors of schizophrenics in Shanghai communities, and to analyze the association between socio demographic characteristics, lifestyles, clinical characteristics and relapse. MethodsA dynamic cohort prospective study design was used in this study. From March 2018 to February 2019, a total of 189 schizophrenics in Xuhui, Hongkou, Changning, Jiading, Songjiang and Baoshan districts were enrolled successively. Baseline questionnaires were conducted through face-to-face interviews at baseline, which contained social demographic information, lifestyle information and clinical information. A follow-up was conducted every 2 weeks for a measurement of PANSS Scale for a total of 6 months. Relapse was assessed by a PANSS score increase of ≥25% from baseline (or an increase of 10 points or more if the baseline score was ≤40 points). Univariate and multivariate Cox regression models were used to analyze the associations between relapse status (assessed by PANSS Scale) and socio demographic characteristics, lifestyles, and clinical characteristics, respectively. ResultsA total of 165 community schizophrenics completed baseline and follow-up surveys, with a loss to follow-up rate of about 12.7%. After exclusion of sociodemographic and clinical information deficits, 132 patients were included in the analysis totally, with an average age of 48.18±12.67 years, among whom 41.67% were male. Totally 33 patients relapsed during the 6-month follow-up period, with a relapse rate of 25.0%. After adjusting for gender, family history, age, employment, education, marital status, smoking, drinking, exercise frequency, medication compliance, insight, social function, violence history, stress recent events, adverse drug reactions and baseline scores of PANSS Scale, risk factors of relapse included the following four factors: age below 40 years (HR=4.47, 95%CI: 1.15-17.40), primary school or below (HR=7.11, 95%CI: 1.54-32.83), unemployed (HR=8.34, 95%CI: 1.78-38.98), and adverse drug reactions (HR=5.02, 95%CI: 1.75-14.37). ConclusionWe should pay attention to the risk factors such as age, education, employment and adverse drug reactions, in order to identify high-risk patients and to conduct timely interventions during the relapse management of schizophrenics in Shanghai community.

2.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-988905

RESUMO

ObjectiveTo explore the effect of paliperidone palmitate treatment on schizophrenic patients in the community. Methods446 schizophrenic patients who used paliperidone palmitate injection were selected in Shanghai. Before and after the treatment, the disease family burden scale, the concise evaluation scale of drug treatment compliance, the VAS100 score of treatment satisfaction, the short form of quality of life measurement scale, and the screening scale of social function defects were used to evaluate the effects of paliperidone palmitate injection. The data were statistically analyzed using SPSS 26.0 software. ResultsAfter using paliperidone palmitate injection, the total score of family burden (13.94±12.17), the score of daily family activities (3.26±2.74), the score of family entertainment activities (2.21±2.30), and the score of family relationship (2.79±2.76) were significantly higher than those before the treatment (14.98±12.64, 3.51±2.88, 2.48±2.38, 3.11±2.87, respectively, all with P<0.05). The scores of the World Health Organization on quality of life brief scale (62.89±11.94) and the medication compliance scale (28.11±5.64) were better than those before treatment (60.67±12.62 and 27.37±6.96, all with P<0.05). Compared with the prior treatment without paliperidone palmitate injection, the number of readmissions after treatment was significantly reduced (P<0.01). ConclusionThe treatment of paliperidone palmitate injection has significant effect, which can effectively reduce the disease family burden of Schizophrenic patients, improve their quality of life, enhance their drug compliance, reduce the readmission rate of patients, ensure long-term treatment effect and promote disease recovery.

3.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-886546

RESUMO

@#Objective    To summarize the experience of surgical treatment of asphyxiating thoracic dysplasia (Jeune syndrome). Methods    A total of 15 patients with asphyxiating thoracic dysplasia from August 2018 to April 2020 in our hospital were retrospectively included. There were 7 males and 8 females, aged 1-25 (8.87±6.71) years. Special steel bars were used to correct the growth direction of the rib and costal cartilage. Meanwhile, the concave and convex deformities of the chest wall on both sides were corrected to increase the chest volume and correct the thoracic deformity. Results    The contour appearance of the chest wall of all patients changed after the operation. The shape was close to normal, and the symptoms of hypoxia were improved. The operation time was 147.73±59.78 min, intraoperative bleeding volume was 105.67±91.90 mL, ICU stay time was 14.20±13.54 d and hospital stay time was 26.00±17.87 d. Eleven patients were directly extubated after the operation, 4 patients underwent tracheotomy and received assisted respiration, and the assisted respiration time was 19, 13, 22 and 12 days, respectively. The postoperative chest circumference was significantly increased, and the blood oxygen saturation was significantly improved. There were 5 patients with cardiac insufficiency, and 3 of them were improved by cardiotonic therapy, 2 of them died of heart failure on the 2nd and 31st day after the operation, respectively. Abdominal distention occurred in 10 patients after operation, and 5 of them were obstinate and eliminated by comprehensive treatment. All patients were followed up. The appearance of thorax was improved obviously and there was no sign of compression in lungs. One 13-year-old patient developed respiratory discomfort 3 months after the operation, and the symptoms were relieved after self-administration of oxygen. A 25-year-old patient developed cardiac insufficiency half a month after the discharge, and the symptoms disappeared after cardiotonic treatment. Four patients took out the steel bars in 13, 13, 15 and 17 months after the operation, respectively. The appearance of thorax remained well after the operation. The imaging examination showed that the position of bone structure was normal, the lung field was clear, and there was no sign of chronic inflammation. Conclusion    This technique is a safe and simple operation method. It can not only eliminate the deformity of chest wall, but also increase the volume of chest obviously. However, the long-term effect needs to be further evaluated.

4.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-745689

RESUMO

Recent studies revealed a number of interesting extrapancreatic actions of sulfonylureas (SUs),and there existed various mechanisms and targets of extrapancreatic effects among different SUs.We try to explore the impact of extrapancreatic effects on the clinical outcome of patients with type 2 diabetes mellitus (T2DM) based on the data from basic and clinical researches,including the effects of SUs on atherosclerosis,cognitive impairment,risk factors associated with abnormal bone metabolism,as well as the safety of treatment.All together,extrapancreatic effects of novel sulphonylureas (e.g.glimepiride) are associated with the improved clinical outcomes and less adverse effects in patients with T2DM.

5.
Mycobiology ; : 207-216, 2019.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-760538

RESUMO

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at 28°C, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ΔAcuD was significantly attenuated in this Zebrafish infection model.


Assuntos
Técnicas de Inativação de Genes , Hifas , Isocitrato Liase , Microinjeções , Mortalidade , Rombencéfalo , Interferência de RNA , Talaromyces , Virulência , Leveduras , Peixe-Zebra
6.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-772091

RESUMO

OBJECTIVE@#To review the experience with Wang procedure for treatment of pectus excavatum in young children.@*METHODS@#The clinical data of 21 children with a mean age of 3.3 ± 1.1 years (ranging from 1.5-6 years) undergoing Wang procedure for pectus excavatum were analyzed. A longitudinal incision (1 to 2 cm) was made in the front of the xiphoid, and two tunnels were created using steel bars beneath the muscles on two sides of the chest wall. The fibrous tissue between the diaphragm and the sternum was dissociated, and the steel wires were sutured through the deformed chest wall. After the steel bar was placed in the tunnels, the wires were pulled and fixed in the middle of the bar, and the incision was sutured.@*RESULTS@#All the operations were performed using 3 wires and 1 steel bar. The operation time was 25 to 51 (38.1 ± 9.6) min with an intraoperative bleeding volume of 5 to 10 (7.1±1.5) mL. The time of hospitalization of the patients ranged from 6 to 10 days (mean 8.1±1.3 days). In all the patients, the incision healed smoothly without serious pain or obvious complications. All the patients were followed up for 1 to 13 months after the operation. During the follow- up, no recess recurred and no such complications as bar displacement or transposition occurred. According to the evaluation criteria after pectus excavatum operation, 13 cases had a total score of 9, and 8 had a total score of 8. The overall effect was satisfactory, and there were no cases rated as basically satisfactory or unsatisfactory.@*CONCLUSIONS@#Wang procedure is a good option for treatment of pectus excavatum in young children.


Assuntos
Criança , Pré-Escolar , Humanos , Fios Ortopédicos , Diafragma , Tórax em Funil , Cirurgia Geral , Fixadores Internos , Duração da Cirurgia , Procedimentos Ortopédicos , Métodos , Estudos Retrospectivos , Parede Torácica , Resultado do Tratamento , Processo Xifoide , Cirurgia Geral
7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-743622

RESUMO

Objective To analyze the effect of stratified nursing intervention based on aspiration risk assessment strategy in ICU mechanical ventilation patients. Methods Totally 156 patients with ICU mechanical ventilation admitted between January 2016 and December 2017 were selected as study subjects, and 78 patients were divided into treatment group and control group by random number table method. The control group was given routine nursing interventions for ICU patients undergoing mechanical ventilation, and the treatment group was combined with stratified nursing interventions based on aspiration risk assessment strategy. Compare the two groups of nurses'knowledge, attitude and practice of aspiration prevention, the complications of patient aspiration, patient satisfaction and other indicators. Results The scores of nurses′knowledge, attitude and practice of prevention of aspiration in the treatment group were (46.32 ± 3.24), (51.41 ± 3.56), and (49.36 ± 4.85), respectively, which were significantly higher than those of the control group (41.25 ± 5.32), (47.12 ± 5.45), (44.42 ± 5.46), The difference was statistically significant (t=4.815, 3.899, 4.002, P<0.05);The rate of aspiration, reflux, asphyxia, aspiration pneumonia in the treatment group was 7.69% (6/78), 19.23% (15/78), 1.28% (1/78), 2.56% (2/78), which were significantly lower than 29.49% (23/78), 39.74% (31/78),12.82% (10/78),20.51% (16/78) in the control group. The difference were statistically significant (χ2=7.892-12.309, P<0.05 or 0.01);Patient satisfaction 97.87% (74/78) was significantly higher than the control group 83.33% (65/78), The difference was statistically significant (χ2=5.347, P<0.05). Conclusion The stratified nursing intervention based on the aspiration risk assessment strategy can help develop nurses′ability in knowledge, attitude and practice of aspiration prevention, reduce the occurrence of complications such as aspiration, and improve patient satisfaction.

8.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-492770

RESUMO

Objective To observe the effect of transcutaneous electrical stimulationon on Jiaji acupoint (Ex-B 05) on the expression of glial fibrillary acidic protein (GFAP), nuclear factor kappa B (NF-κB) and interleukin-6 (IL-6) in rats with spinal cord injury (SCI). Meth-ods Ninety healthy adult Sprague-Dawley rats were randomly divided into normal group (group A, n=30), transcutaneous electrical stimula-tion treatment group (group B, n=30) and control group (group C, n=30). Allen's method was used to establish the model of acute SCI in T9. All of them were assessed with Basso-Beattie-Bresnahan (BBB) scale and Slanting Board Test, and the expression of GFAP, NF-κB and IL-6 of the spinal cord were detected with immunohistochemistry one, three and seven days after operation. Results The results of BBB scale and Slanting Board Test were better in group B than in group C three and seven days after SCI (t>3.349, P20.815, P<0.001). Conclusion Transcutane-ous electrical stimulation can inhibit the inflammation and the expression of GFAP in spinal cord after SCI, and improve the motor function in rats after SCI.

9.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-487340

RESUMO

Objective To investigate the clinical efficacy of triple method of heat-sensitive point moibustion and Taiji acupuncture plus massotherapy in treating facial spasm.Method Sixty patients with facial spasm were randomly allocated to treatment and control groups, 30 cases each. The treatment group received triple method of heat-sensitive point moibustion and Taiji acupuncture plus massotherapy and the control group, conventional acupuncture plus suspended moxibustion with moxa stick. After three courses of treatment, the clinical therapeutic effects on spasm (severity and frequency) were compared between the two groups. Result The total efficacy rate and the cure and marked efficacy rate for spasm (severity) were 86.7% and 43.3%, respectively, in the treatment group and 66.7% and 26.7%, respectively, in the control group; there were statistically significant differences between the two groups (P<0.01). The total efficacy rate and the cure and marked efficacy rate for spasm (frequency) were 83.3% and 46.7%, respectively, in the treatment group and 66.7% and 30.3%, respectively, in the control group; there were statistically significant differences between the two groups (P<0.01).Conclusion Triple method of heat-sensitive point moibustion and Taiji acupuncture plus massotherapy is an effective way to treat facial spasm.

10.
Zhonghua Yi Xue Za Zhi ; 95(13): 969-72, 2015 Apr 07.
Artigo em Chinês | MEDLINE | ID: mdl-26506704

RESUMO

OBJECTIVE: To evaluate the clinical outcomes and operative techniques of microsurgical resection of lumbar intraspinal tumors through paraspinal approach by percutaneous tubular retractor system. METHODS: A retrospective study was conducted to analyze 21 patients with lumbar intraspinal tumors between November 2011 and February 2014, including Schwannoma (n = 19) and meningioma (n = 2) without lumbar instability on preoperative images. The length of tumors was 0.6-2.0 cm. Tracheal intubation anesthesia was performed prior to microsurgery using percutaneous tubular retractor system through paraspinal approach. Operative duration, blood loss volume, postoperative wound pain duration and hospital stay were analyzed. Creatine phosphokinase (CPK-MM) level was recorded at 1 day preoperatively, 1 day, 3 days and 5 days postoperatively. The scores of Japanese Orthopedic Association (JOA) and visual analog scale (VAS) were analyzed at 1 day preoperatively, 1, 3, 5 days and 6 months postoperatively to evaluate the function status of spinal cord. Computed tomography ( CT) three-dimensional reconstruction of lumbar vertebrae was performed at 1 week postoperatively. Magnetic resonance imaging ( MRI) plain scan and enhanced scan of lumbar vertebrae were conducted preoperatively, 1 week and 6 months postoperatively. RESULTS: Complete removal of tumors was achieved in all patients without the injuries of spinal cord or nerve root. Postoperative scores of JOA and VAS improved versus preoperative ones (P < 0.05). Level of CPK-MM increased 1 day postoperatively and declined to preoperative level at 5 days postoperatively. And th difference was not statistically significant (P < 0.05). Neither residual tumor nor tumor recurrence was detected by MRI plain and enhanced scans. No postoperative spinal instability was identified by CT three-dimensional reconstruction. And no spinal deformity occurred during a follow-up period of 6-28 months. CONCLUSION: Microsurgical resection of lumbar intraspinal tumors using percutaneous tubular retractor system through paraspinal approach minimizes the injuries of paraspinal muscles, facet joint, spinous process and ligaments. This technique offers the advantages of mini-invasiveness, shorter hospital stay, rapid recovery and preserved postoperative spinal stability.


Assuntos
Microcirurgia , Neoplasias da Coluna Vertebral , Humanos , Vértebras Lombares , Região Lombossacral , Imageamento por Ressonância Magnética , Período Pós-Operatório , Recidiva , Estudos Retrospectivos , Tomografia Computadorizada por Raios X , Articulação Zigapofisária
11.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-815211

RESUMO

OBJECTIVE@#To determine the factors associated with the eff ect of steroid on adult primary nephrotic syndrome.@*METHODS@#The general information, laboratory examination and renal pathological type of 425 patients with primary nephrotic syndrome were retrospectively analyzed.@*RESULTS@#Th ere were significant differences in the response to steroid among the pathological types of minimal change disease, focal segmental glomerulosclerosis and IgA nephropathy. Th e patients in the age of 14-24 years old showed the strongest response to steroid (P0.05), while there was significant difference in urine protein in 24 hour quantitation in the non-steroid resistance group between pre- and post-treatment (P<0.05).@*CONCLUSION@#Pathological types and ages of the patients are related to the steroid curative effect. The decrease in IgA probably affects the effect of steroid on primary nephrotic syndrome.


Assuntos
Adolescente , Adulto , Humanos , Adulto Jovem , Glomerulonefrite por IGA , Tratamento Farmacológico , Glomerulosclerose Segmentar e Focal , Tratamento Farmacológico , Rim , Nefrose Lipoide , Tratamento Farmacológico , Síndrome Nefrótica , Tratamento Farmacológico , Proteinúria , Estudos Retrospectivos , Esteroides , Usos Terapêuticos , Urinálise
12.
China Pharmacy ; (12): 4504-4505, 2015.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-501185

RESUMO

OBJECTIVE:To establish a rapid method for the concentration determination of lamotrigine (LTG) in human se-rum. METHODS:HPLC was adopted after serum samples treated with methanol. The column was Hypersil BDS C18(200 mm×4.6 mm,5μm)with the mobile phase of methanol-KH2PO4(45∶55,V/V)at the flow rate of 1 ml/min,the detection wavelength was 250 nm,and the temperature was 35℃. RESULTS:The linear range of LTG was 0.5-30.0 μg/ml(r=0.999 9). The RSDs of intra-day and inter-day were all less than 5% and the recovery was 101.96%-102.74%. CONCLUSIONS:The method is simple,accurate and fast,and suitable for the monitoring of LTG serum concentration.

13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-940062

RESUMO

@#Objective To observe the effect of proanthocyanidin on the expression of brain- derived neurotrophic factor (GFAP) and brain-derived neurotrophic factor (BDNF) in rats with spinal cord injury (SCI). Methods 48 healthy adult Sprague-Dawley rats were randomly divided into proanthocyanidin treatment group (group A, n=24) and control group (group B, n=24). Allen's method was used to establish the model of acute spinal cord injury in T9. 1, 3 and 7 days after operation, 8 rats in each subgroup were assessed with Basso-Beattie- Bresnahan (BBB) scale and Slanting Board Test, the serum malondialdehyde (MDA) and superoxide dismutase (SOD) were detected, and the expression of GFAP and BDNF of the spinal cord were detected with immunohistochemistry. Results The results of BBB scale and Slanting Board Test were better in group A than in group B 3 and 7 days after SCI (P<0.05). The levels of SOD and MDA were better in group A than in group B 1, 3 and 7 days after SCI (P<0.05). The expression of GFAP was lower, and the expression of BDNF was higher in group A than in group B all the time points after SCI (P<0.01). Conclusion Proanthocyanidin can inhibit the lipid peroxidation and the expression of GFAP in spinal cord after SCI, stimulate the synthesis of endogenous BDNF, and improve the motor function in rats after SCI.

14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-476880

RESUMO

Objective To observe the effect of proanthocyanidin on the expression of brain-derived neurotrophic factor (GFAP) and brain-derived neurotrophic factor (BDNF) in rats with spinal cord injury (SCI). Methods 48 healthy adult Sprague-Dawley rats were ran-domly divided into proanthocyanidin treatment group (group A, n=24) and control group (group B, n=24). Allen's method was used to estab-lish the model of acute spinal cord injury in T9. 1, 3 and 7 days after operation, 8 rats in each subgroup were assessed with Basso-Beat-tie-Bresnahan (BBB) scale and Slanting Board Test, the serum malondialdehyde (MDA) and superoxide dismutase (SOD) were detected, and the expression of GFAP and BDNF of the spinal cord were detected with immunohistochemistry. Results The results of BBB scale and Slanting Board Test were better in group A than in group B 3 and 7 days after SCI (P<0.05). The levels of SOD and MDA were better in group A than in group B 1, 3 and 7 days after SCI (P<0.05). The expression of GFAP was lower, and the expression of BDNF was higher in group A than in group B all the time points after SCI (P<0.01). Conclusion Proanthocyanidin can inhibit the lipid peroxidation and the ex-pression of GFAP in spinal cord after SCI, stimulate the synthesis of endogenous BDNF, and improve the motor function in rats after SCI.

15.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-254460

RESUMO

<p><b>OBJECTIVE</b>To explore the impact of Line-1 methylation on clinical features of non-small cell lung cancer and its connection with smoking and other living habits.</p><p><b>METHODS</b>Pyrosequencing was used to determine the extent of Line-1 methylation in cancer and adjacent tissues derived from 197 patients with primary non-small cell lung cancer. Non-conditional logistic regression analysis was performed to correlate the level of Line-1 methylation with clinical features and living habits of the patients.</p><p><b>RESULTS</b>Line-1 methylation for cancer tissue and adjacent tissue has measured 68.20±11.63 and 78.90±2.09, respectively (P < 0.01), and has been associated with TNM staging, smoking history and histopathological types.</p><p><b>CONCLUSION</b>Lung cancer tissue Line-1 methylation level is closely related with clinical features and smoking. There is also a correlation between histopathological types of lung cancer and relative hypomethylation of Line-1.</p>


Assuntos
Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Pulmonar de Células não Pequenas , Genética , Metabolismo , Patologia , Metilação de DNA , Elementos Nucleotídeos Longos e Dispersos , Neoplasias Pulmonares , Genética , Metabolismo , Patologia
16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-291711

RESUMO

<p><b>OBJECTIVE</b>To explore the incidence and genotypes of glucose-6-phosphate dehydrogenase (G6PD) gene mutations among infertile patients in Shenzhen.</p><p><b>METHODS</b>DNA samples from 851 infertile patients were tested for 25 G6PD gene mutation sites using a multiplex SNaPshot assay.</p><p><b>RESULTS</b>The incidence of G6PD gene mutations among infertile patients in Shenzhen was 17.63%. Male and female abnormal rates were 15.13% and 20.09% respectively. Most of the female abnormal cases were heterozygotes. Mutations involved 11 haplotypes in 10 sites. 1311C> T/IVS-11 93T> C was the most common mutation, accounting for 72.00% (108/150) abnormal cases. Forty three cases of missense mutations were detected, including 19 cases of 1376G> T, 9 cases of 1388G> A, 5 cases of 95A> G and 871G> A/1311C> T/IVS-11 93T> C, 1 case of 202G> A, 835A> T, 1360C> T, 1376G> T and 392G> T/1311C> T/IVS-11 93T> C.</p><p><b>CONCLUSION</b>The incidence of G6PD gene mutations among infertile patients in Shenzhen was high and the mutation types were various. Therefore, the G6PD deficiency genetic screening should be performed prior to assisted reproduction. This investigated results provided valuable basic data for genetic counseling, preimplantation genetic diagnosis and prenatal diagnosis.</p>


Assuntos
Feminino , Humanos , Masculino , Povo Asiático , Genética , China , Epidemiologia , Análise Mutacional de DNA , Frequência do Gene , Genótipo , Glucosefosfato Desidrogenase , Genética , Haplótipos , Incidência , Infertilidade Feminina , Etnologia , Genética , Infertilidade Masculina , Etnologia , Genética , Mutação
17.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-389943

RESUMO

Objective To investigate the expression of exogenous gene transferred by piggyBac (PB) transposon in various gynecological malignant cell lines and reveal its potential application of gene therapy in gynecological cancer.Methods Amplified herpes simplex virus thymidine kinase (HSV-tk) gene coding region by PCR and integrated it into PB expression vector, PB[Act-RFP]DS, for reconstructing PB[Act-RFP, HSV-tk]DS (pPB/TK).By using different transfection reagents: FuGENE HD, jetPEI, lipofectamine 2000, pPB/TK together with helper plasmid Act-PBase were cotransfected into four mostly common gynecological malignant tumor cell lines HeLa, JEG-3, SKOV3 and HEC-1B.The mRFP1 report gene expressions was observed and detected by fluorescence microscope and flow cytometry to analyze transfection efficiency.The expressions of HSV-tk and mRFP1 gene were detected by reverse transcription PCR (RT-PCR).The cytotoxic effect of various concentration of pro-drug ganciclovir (GCV) for transfected cells was detected by methyl thiazole tetrazolium assay.The transfected cells were positive sorted by flow cytometry and limiting diluted to obtain the stable transfected cell line.The insertion sites of foreign gene tranferred by PB transposon in genome were analyzed by inverse PCR.Results (1) Double digests analysis and sequences test demonstrated that pPB/TK vector was reconstructed successfully.(2) Using three different transfective reagents, PB trausposon transferred HSV-tk gene and mRFP1 gene into HeLa, HEC-1 B, SKOV3 and JEG-3 cell efficiently, and the transfection efficiency of pPB/TK for the same cell was different by using different transfective reagents; in Hela cell, the transfection efficiency of FuGENE HD [(78.7 ± 9.2) %]was higher than that of lipofectamine 2000 [(54.1 ± 11.4) %]and jetPEI [(46.5 ± 7.4) %, all P < 0.05] ; using the same transfective reagent, the transfection efficiency of pPB/TK was also different on various cell lines, using FuGENE HD, the transfeetion efficiency of pPB/TK on HeLa, JEG-3 and SKOV3 cell was (78.7 ± 9.2) %, (74.4 ± 8.9) % and (83.2 ± 9.7) % respectively, which all were higher than that on HEC-1B [(39.5 ± 8.7) %, P < 0.05] .(3) RT-PCR showed that there were the mRNA expression of HSV-tk and mRFP1 in all cell lines.(4) 50% inhibitory concentration of GCV for transfected cells, HeLa, JEG-3, SKOV3 and HEC-1B, was 1.29, 3.35, 0.09 and 13.28 μg/ml respectively.Inhibitory effect of GCV (10 μg/ml) on SKOV3 transfected with pPB/TK was (86 ± 9) %, which was superior to that transfected with pORF-HSVtk alone [(52 ± 12)%, P < 0.05] .(5) The insertion sites of PB transposon in the target cells genome were located at TTAA sites, mRFP1 expression still could be detected in three months after transfected.Conclusions PB transposon could transfer exogenous gene into various gynecological malignant cells, which could integrated into genome and obtain a long-term and stable expression.It is expected that PB transposon may supply a more efficient and safer transgene technology platform for gene therapy in gynecological cancer.

18.
Chinese Journal of Dermatology ; (12): 810-813, 2008.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-397502

RESUMO

Objective To develop a multiple PCR-based reverse line blot hydrization assay (mPCR-RLB)to simutaneously detect several STD pathogens:Neisserria gonorrhoeae,Chlamydia trachomatis,Ureaplasma urealyticum,U.parvum,Mycoplasma genitalium,M,hominis and Trichomonas vaginalis.Methods Seven pairs of biotin-labelled primer were designed and synthesized to target the 16S rRNA-23S rRNA intergenic spacer regions of Neisserria gonorrhoeae,Chlamydia trachomatis,Mycoplasma,and repetitive DNA sequence of Trichomonas to identify and subtype thesc pathogens.DNA was extracted from the referrence strains of seven pathogens and used as templates.mPCR was performed to simutaneously amplify the target regions of these pathogens.Then,the biotin-labelled amplicons were hybridized with membrane-bound specific oligonucleotide probes followed by the detection of bound amplicons with chemiluminescence assay.Serially diluted plasmids containing the target genes of pathogens were amplified with this method to detect its sensitivity.Two-hundred and eleven specimens,including 104 male urethral swabs and 107 female cervical swabs,were collected from the STD clinic of Wuhan First Hospital;mPCR-RLB and single-primer PCR were performed.For specimens with inconsistent results,nested PCR was performed to confirm the results.Results The assay sensitively and specifically identified referrence strains of the tested pathogens.The detection limit of mPCR-RLB was 100 copies for all the pathogens.Of the 211 clinical specimens,2.8%(6/21)were negative for single-primer PCR,but positive for mPCR-RLB,and nested-PCR results were consistent with those of mPCR-RLB.Conclusion mPCR-RLB is a sensitive,specific and rapid method for the detection of STD pathogens from clinical specimens.

19.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-566138

RESUMO

0.05).Conclusions In vivo ischemia models,the suitable dose GM can reduce the expression of iNOS,inhibit the cell apoptosis and protect the nerve cell.

20.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-564389

RESUMO

Aim To investigate the inhibition of cell proliferation and the expression of PCNA gene of human glioma U251 by elemene.Methods Normal control and treatment group with elemene in human glioma U251 were studied, then the inhibition effect was examined by MTT methods and the density of half death was worked out (IC50). The expression of PCNA protein and gene was examined by immunohistochemical method and RT-PCR in different action time or at different concentrations.Results Elemene had a marked antineoplastic effect on glioma U251 with the dependence of dose and time,IC50 was 0.062 g?L-1. Elemene exhibited the expression of PCNA protein and gene,which could be examined by immunohistochemical method and RT-PCR. The expression of PCNA mRNA was lowered with the increase of drug concentration and action time.Conclusion Elemene inhibited the expression of PCAN gene,induced the inhibition of cell proliferation,and had a marked antiproliferative effect on glioma cells.

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