1.Preparation and identification of polyclonal antibody against gastric cancer-related protein GCRG224
Yinqiao WU ; Gangshi WANG ; Mengwei WANG
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective To prepare a polyclonal antibody against gastric cancer-related protein GCRG224.Methods The thioredoxin/GCRG224 fusion protein was expressed in E.coli.A polyclonal antibody against GCRG224 was obtained by immunizing a rabbit with the purified GCRG224 protein.The titer and specificity of the antibody were determined by ELISA and Western-blot,respectively.Results The thioredoxin/GCRG224 fusion protein with relative molecular mass of 16.8kD was over-expressed in E.coli.The purity of expressed products directly purified from a denaturing polyacrylamide gel was about 100%.The polyclonal antibody against GCRG224 was obtained.The ELISA titer of antiserum against GCRG224 was about 1∶256 000.Western blot analysis showed that the antiserum could bind to the expressed fusion protein specifically.Conclusion The polyclonal antibody against GCRG224 has been successfully prepared,which lays the foundation for further study on the biological function and the possible role of the GCRG224 in the development of gastric carcinoma.
2.Expression of gastric cancer related gene GCRG213 in E.coli and purification of recombinant protein
Yinqiao WU ; Gangshi WANG ; Mengwei WANG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To express gastric cancer related gene GCRG213 by using thioredoxin fusion expression system, and to prepare human GCRG213 fusion protein. Methods GCRG213 cDNA with complete open reading frame was amplified by PCR from plasmid pGEM-T, and then was cloned into thioredoxin fusion expression vector pET102/D-TOPO. The recombinant plasmid was further transformed into E.coli BL21 strain. After induction with IPTG, the thioredoxin/GCRG213 fusion protein was expressed in E.coli. The product was obtained by means of direct purification from a denaturing polyacrylamide gel. Results SDS-PAGE analysis showed the thioredoxin/GCRG213 fusion protein with relative molecule mass of 29.4kDa was highly expressed. The thin layer gel scanning analysis showed that the yield of GCRG213 fusion protein was 28.7% of the total bacterial protein. The product was obtained with a purity of about 100% by means of direct purification from a denaturing polyacrylamide gel. Conclusion The thioredoxin/GCRG213 fusion protein was successfully expressed in E.coli and the product with high purity was obtained, which laid the foundation for the function research and antibody preparation hereafter.
3.Cloning and primary expression analyses of down-regulated cDNA fragment in human gastric cancer
Gangshi WANG ; Mengwei WANG ; Weidi YOU ; Hong LI
Chinese Journal of Medical Genetics 2001;18(1):43-46
Objective To identify human gastric cancer related genes. Methods Specimens of paired tumor, paratumor and normal gastric mucosa tissues were collected from five patients (male 3, female 2, with average age 48.8±18.1 years) who suffered from stomach antrum adenocarcinoma. Total RNA samples were extracted from these specimens, then studied by fluorescent differential display reverse transcription polymerase chain reaction (DDRT-PCR) analysis. The differentially expressed bands of interest were recovered, purified and cloned, then they were analyzed by sequencing, Northern blot and RT-PCR. Through BLAST, the sequencing results were compared with GenBank database for homology analysis.Results One of the interesting cDNA bands expressed much lower in all five tested tumor samples than in their normal and paratumor counterparts. This band was named W4. Northern blot analysis showed a consistent result with that of DDRT-PCR. W4 was cloned into pGEM-T easy vector. Sequence analysis showed that W4 consists of 712bp, this sequence was named W44.BLAST analysis revealed that W44 has extremely low sequence identity with any genes from GenBank and any sequences from EST database. This sequence data was submitted to GenBank with accession No. AF150631. RT-PCR analysis showed that W4 was expressed much lower in 11/15 gastric cancer tissue than in paratumor and normal samples. Conclusion A novel cDNA sequence related to human stomach adenocarcinoma was identified.
4.A comparative study of using Brix meter versus ultrasonic monitoring of gastric residual volume in patients with enteral nutrition
Yu DING ; Gangshi WANG ; Ying ZHANG ; Huiru HOU
Chinese Journal of Internal Medicine 2017;56(12):940-942
To investigate the accuracy and feasibility of Brix value on monitoring gastric residual volume (GRV) in patients with enteral nutrition. Fifty patients with enteral nutrition via nasogastric tube were enrolled. The GRV was measured by both ultrasonography and Brix value. The results were compared according to the methods. The Pearson correlation coefficients showed that GRV measured by these two ways was positively correlated (r=0.986, P<0.05). Moreover paired sample t-test showed that the discrepancy was not statistically significant (P>0.05) between different measurements. The consistency was analyzed by Bland-Altman graph, showing that the two measurements were consistent. Brix value is recommended to measure GRV due to its convenience and easy operation.
5.A lamin-like protein gene is down-regulated in human gastric cancer.
Gangshi WANG ; Mengwei WANG ; Benyan WU ; Weidi YOU
Chinese Journal of Medical Genetics 2003;20(2):119-122
OBJECTIVETo clone human gastric cancer related gene and to analyze its expression profile in gastric mucosal tissues.
METHODSPaired tumor, paratumor and non-tumor specimens from 7 gastric adenocarcinoma patients (male 4, female 3, with average age 51 +/- 18 years) were studied by means of fluorescent differential display reverse transcription polymerase chain reaction (DDRT-PCR). The differentially expressed cDNA bands of interest were cloned and analyzed by Northern blot and in situ hybridization. Thirty cases (male 23 female 7 with average age 59 +/- 8 years) of paired paraffin-embedded gastric tumor and non-tumor tissues were used in in situ hybridization analysis.
RESULTSA gene expressed much lower in 6 out of 7 tested tumor samples than in their normal and paratumor counterparts was identified. It was named GCRG123. Northern blot analysis confirmed the differential expression. Human multiple tissue Northern blot analysis showed that GCRG123 expressed in various adult human tissues including thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocyte. Sequence analysis revealed that GCRG123 (GenBank accession number AF454554) was a lamin like protein gene. It had one open reading frame which consisted of 49 amino acids (GenBank accession number AAL61668.1). In situ hybridization analysis showed a high GCRG123 expression level in normal gastric epithelium and pylori glands, but low expression level in tumor as well as dysplasia and most intestinal metaplasia at the paratumor regions.
CONCLUSIONA lamin-like protein gene was identified in human gastric mucosa, it is down-regulated in gastric cancer and its precancerous leisions.
Adult ; Aged ; Blotting, Northern ; Cloning, Molecular ; DNA, Neoplasm ; chemistry ; genetics ; Down-Regulation ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; In Situ Hybridization ; Lamins ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; genetics ; metabolism ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics
6.Cognition and Education of Knowledge of Medical Ethics in Geriatric Ward Medical Staffs
Yue LYU ; Tianzhi LI ; Huiru HOU ; Yuan GONG ; Weimin DONG ; Gangshi WANG
Chinese Medical Ethics 2018;31(1):124-127
Objective:To investigate the mastery and application of medical ethics basic knowledge in geriatric ward medical staffs,and the way which they receive relevant education. Methods:From November 2016 to January 2017,240 questionnaires about medical ethics were distributed among medical staffs in clinical department in the Chinese PLA General Hospital,among them,30 were medical groups and 210 were nursing groups;and a total of 233 valid questionnaires were collected. The nurses were divided into geriatric ward nursing group and non - geriat-ric ward nursing group,according to whether they work in the geriatric wards or not. Results:Only 2. 14% and 7. 72% participants fully and correctly answered the core concepts and basic principles of medical ethics,respec-tively. The rate of informed consent in medical staffs in geriatric group was significantly higher than that of non -geriatric group (92. 66% vs 82. 11% ,P = 0. 02);the patient' s privacy protection in geriatric nursing group was significantly higher than that of non - geriatric nursing group (92. 66% vs 31. 57% ,P < 0. 001). 28. 75% of the respondents received medical ethics continuing education curriculum,and the proportion of correctly receiving ex-pectable death in these medical staffs (71. 40% ) was significantly higher than that of other education pathway groups (36. 17% ). Conclusion:Medical staffs lack more knowledge on medical ethics. Clinical work in geriatric medicine has higher demand for medical ethics knowledge,and continuing medical education can help to improve the cognitive level of medical ethics in medical staffs.
7.The interRAI acute care for comprehensive geriatric assessment in elderly emergency patients
Hailan ZHU ; Fansen MENG ; Yunjuan LIN ; Haiyan SHI ; Liyuan WANG ; Haiyan XIE ; Xiaomei PEI ; Gangshi WANG
Chinese Journal of Geriatrics 2020;39(11):1292-1296
Objective:To evaluate dynamic changes of functional status in elderly emergency inpatients by using the interRAI Acute Care for Comprehensive Geriatric Assessment(interRAI AC-CGA), and to analyze whether assessment results are helpful for clinical decision-making.Methods:Elderly patients(aged 60 years and older)who were hospitalized in the Acute Care Unit of PLA General Hospital from October 2017 to March 2018 were included.Each patient received the interRAI AC-CGA instrument-based assessment at admission(day 1), on day 7 and day 28 after admission.Analysis of collected data was conducted using interRAI AC-CGA scales, geriatric syndrome screening and risk identification of adverse outcomes, in order to examine dynamic changes of functional status in patients at and after admission and to explore the relationship between functional status at admission and outcomes(at discharge).Results:Thirty-four patients were evaluated, including 26 males and 8 females, aged from 60 to 101 years, with a mean age of(84.2±10.1)years.Patients had a variety of functional abnormalities at admission and during hospitalization, in which the ability to perform activities of daily living(ADL)and instrumental activities of daily living(IADL)decreased significantly.Compared with evaluation results on day 1, the incidences of functional abnormalities on day 7 and day 28 declined, and there was improvement in transient depression, pain and falls( χ2=6.298, 6.027 and 7.766, P=0.043, 0.049 and 0.021). After evaluation on day 28, 16 patients were discharged and 18 remained hospitalized(including 2 deaths). The rate for most abnormality was lower in the discharge group than in the continued hospitalization group, and the rates for abnormal communication and falls were lower in the discharge group than in the continued hospitalization group( χ2=6.349 and 5.443, P=0.019 and 0.030). Age(≥85 years old)had no effect on discharge.There was no significant difference in hospital discharge rate between the advanced age group(≥85 years old)and the elderly group(60-84 years old)(40.0% or 8/20 vs.57.1% or 8/14, χ2=0.971, P=0.324). Conclusions:The interRAI AC-CGA instrument can systematically and dynamically estimate functional changes in elderly emergency inpatients.Functional status at admission is related to prognosis(performance at discharge)of patients.
8. Treatment of postprandial discomfort syndrome in the elderly: a multi-centered prospective randomized controlled clinical study
Gangshi WANG ; Le XU ; Hongtan CHEN ; Liping SHI ; Minjing HUANG ; Ling XI ; Lishu XU ; Fen WANG ; Hongyi LI ; Shu LI ; Yijun ZHANG ; Shiyun TAN ; Rutao HONG ; Nonghua LYU ; Mei YE ; Huatian GAN ; Miao LIU ; Benyan WU
Chinese Journal of Internal Medicine 2020;59(2):117-123
Objective:
To evaluate the efficacy and safety of Oryz-Aspergillus enzyme and pancreatin tablets (Combizym®) in the treatment of postprandial distress syndrome (PDS) in the elderly, compared with gastrointestinal motility drugs.
Methods:
A prospective randomized controlled trial was designed and registered in the China Clinical Trials Registry (ChiCTR-IPR-16008185). The elderly patients with PDS were randomly divided into three groups, including Mosapride group with Mosapride citrate tablets 5 mg 3 times per day for 2 weeks; Combizym® group with Combizym tablets 244 mg 3 times per day for 2 weeks; combined treatment group with both drugs and same doses for 2 weeks. The modified Nepean dyspepsia index (NDSI) score, discomfort intensity score and PDS score were calculated on patients before treatment, at the end of first and second week of treatment, as well as 4 weeks after treatment finished, respectively. Adverse effects were evaluated.
Results:
A total of 323 patients from 16 tertiary hospitals in China were enrolled in this study. Among them, 105 patients were in Mosapride group, 109 in Combizym® group and 109 in combined treatment group. There were 148 males (45.8%) and 175 females (54.2%) with median age 71.4±9.0 years (60-100 years). Baseline characteristics of three groups were comparable. After treatment, the NDSI scores in three groups all decreased significantly (