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Objective:To evaluate the clinical status, and analyz obstacles and facilitators for perioperative deep vein thrombosis prevention of brain neoplasms based on the Ottawa model of research use (OMRU).Methods:A total of 93 patients with brain tumors who were admitted to the Department of Neurosurgery, the First Affiliated Hospital of Nanjing Medical University from April to May 2021 and 33 nurses in the neurosurgery ward and operating room neurosurgery special group were selected as the baseline review subjects by convenience sampling. Based on the framework of evidence-based continued quality improvement of Fudan University, we searched BMJ Best Practice, UpToDate, The Joanna Briggs Institute Library, International Guideline Library, American Guideline Network, Scottish Intercollegiate Guideline Network, National Institutes for Health and Clinical Technology Optimization, Medline, Medlive, China National Knowledge Infrastructure, VIP, Wanfang and SinoMed according to the '6S' evidence pyramid from inception to January 1, 2021 for all clinical decisions, recommended practices, best practice information, evidence summary, guidelines and expert consensus on venous thrombosis assessment, prevention, screening, nursing and health education. The best evidence was summarized, and the final review indicators were formulated through two rounds of expert correspondence. According to the results of baseline review, barriers and facilitators were analyzed, and countermeasures were developed guided by OMRU.Results:A total of 19 best evidences were included, and 34 review indicators were developed in this study. Among them, only 4 indicators had a compliance rate of 100%, 18 ones had a compliance rate of 0, and the other 12 ones had a compliance rate of 6.5%-97.8%. A multi-factor analysis of the review results showed that the main obstacles of evidence implementation were the feasibility and comprehensibility at evidence level, the lack of knowledge and heavy workloads at the potential practitioner level, insufficient education materials, trainings and preventive equipment at system level. Furthermore, the reliable sources of evidence at evidence level, supports from practitioners at the potential practitioner level and system resources (such as training, national and hospital policies, etc.) at system level may contribute to the clinical application of evidence.Conclusions:There was still a huge gap between the best evidence and clinical practice. The obstacles and facilitating factors in evidence transformation should be evaluated scientifically and comprehensively, and corresponding countermeasures should be given to promote the application of evidence in clinical practice.
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Objective:To analyze the results of plague surveillance in Qinghai Province from 2011 to 2020, master the epidemic situation in recent years, and provide scientific basis for prevention and control of the plague in the future.Methods:The human plague epidemic data (from the human case database of Qinghai Institute for Endemic Disease Prevention and Control) and animal plague epidemic data (from plague monitoring data and plague focus survey data of Qinghai Province) from 2011 to 2020 were collected and analyzed with descriptive epidemiological methods, including human plague epidemic, animal plague epidemic regional distribution, host animal monitoring results, pathogenic monitoring results and serological monitoring results.Results:From 2011 to 2020, there was a human plague epidemic in Qinghai Province, which was infected due to the infection of a middle finger of the right hand that was accidentally scratched when peeling marmots, and Yersinia pestis was isolated from heart, liver, lung, lymph node puncture fluid, tracheal secretion and throat swab samples of the deceased. There were 16 animal plague epidemics and endemic areas were distributed in Haixi Prefecture, Yushu Prefecture and Haibei Prefecture, among which the animal plague epidemic was the most prevalent in Haixi Prefecture, with 13 outbreaks in recent 10 years. According to the monitoring of host animals, the main host animal was the Himalayan marmot, with an average density of 0.07/hm 2. Pathgenic monitoring showed that 31 strains of Yersinia pestis were isolated, of which 27 strains were isolated from Haixi Prefecture. The host animals of Yersinia pestis were mainly Himalayan marmot, accounting for 77.42% (24/31) of the total. Serological monitoring showed that 66 plague F1 antibody positive sera were detected, of which 43 were dog positive sera; the Himalayan marmot took the second place, 20. Conclusion:From 2011 to 2020, the animal plague in Qinghai Province has continued for many years, with some areas showing an active trend, and the overall situation of plague prevention and control is severe.
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Objective:To explore the effect of application in patients undergoing general anesthesia prone surgery on basis of observation of ocular complications and focus on preventive care.Methods:One hundred patients who underwent general anesthesia prone surgery before the start of the ocular complications focused preventive care exploration activity (January to May 2019) were set as the control group. After the ocular complications focused preventive care exploration activity was launched (June to October, 2019) 100 cases of general anesthesia prone surgery in our hospital were set as the experimental group, and the observation indexes after the two groups were compared.Results:The satisfaction scores of nurses and patients in the observation group were (9.40 ± 0.57) and (9.30 ± 0.56) respectively, while those in the control group were (7.51 ± 0.88) and (7.09 ± 1.10) respectively, the difference between the two groups was statistically significant ( t values were 18.013 and 13.063, all P < 0.05). The incidence rates of conjunctival congestion, edema and exposure keratitis in the observation group were 4.00% (4/100), 2.00% (2/100) and 1.00% (1/100), respectively, while those in the control group were 12.00% (12/100), 10.00% (10/100) and 9.00% (9/100), respectively. the difference between the two groups was statistically significant ( χ 2 values were 4.348, 5.674, 6.737, all P< 0.05). Conclusion:The focus on preventive care of ocular complications in patients undergoing general anesthesia prone surgery can significantly reduce the risk of ocular complications in this type of patients, and it is well recognized by both patients and patients.
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Objective:To study the clinical characteristics and treatment measures of postoperative bleeding in hyperparathyroidism surgery.Methods:A total of 56 patients with secondary hyperparathyroidism in Shunde Hospital from November 2014 to November 2018 were underwent parathyroidectomy, from which 5 cases developed postoperative hemorrhage were collected and analyzed.Results:Among the 56 patients, bleeding after operation were found in 5 cases(5/56, 8.93%), including hemorrhage of anterior jugular vein in 1 case, hemorrhage of thyroid wound after thyroid mass resection in 1 case, hemorrhage of anterior jugular muscles in 3 cases, hemorrhage of cricothyroid muscle in 1 case.All patients were cured by treatment in time without death.Conclusion:Early diagnosis and proper treatment are crucial for reducing the complications of post-parathyroidectomy bleeding.
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Objective To investigate the association between diabetic retinopathy ( DR) and atherosclerosis cardiovascular disease ( ASCVD) .Methods Clinical data of 654 patients with type 2 diabetic mellitus (T2DM) were collected from a cross sectional , population based survey on chronic diseases and risk factors which was conducted in Beijing Changping district from July 2010 to March 2011.Among 654 T2DM patients, there were 73 patients with ASCVD (ASCVD group) and 581 patients without ASCVD ( non-ASCVD group ) .The association between DR and ASCVD was analyzed .Results Patients with ASCVD had significantly older age [58.5(53.9,65.9) years], more female sex[52(71.2%)], higher proportion of ASCVD history [45(61.6%)], higher levels of PG 2 h[16.26(11.08,19.20) mmol/L], HbA1c[7.20(6.55,8.85)%], systolic pressure [151(133,165) mmHg(1 mmHg＝0.133 kPa)] and lower eGFR[87.2(75.0,103.0) ml· min-1· 1.73 m-2] than non-ASCVD patients[52.4(46.5,58.3) years, Z＝-5.86, P＝0.00; 307(52.8%),χ2＝-8.86, P＝0.00; 256(44.1%),χ2＝8.07, P＝0.01; 13.10(8.99,17.93) mmol/L, Z＝-2.35, P＝0.02; 6.70(6.00, 7.90)%, Z＝-3.33, P＝0.00; 143(131,158) mmHg, χ2＝-2.28, P＝0.02; 94.6(84.8,106.3) ml· min-1· 1.73 m-2, Z＝-3.47, P＝0.00].The trend to develop DR in ASCVD group was significantly higher than that in non-ASCVD group [19.2%(14/74) vs.8.3%(48/581), χ2＝9.01, P ＝0.00] .DR was an independent statistical indicator of the presence of ASCVD [ OR ( 95%CI ): 2.64 ( 1.37 -5.06 ), P ＝0.00 ] . Furthermore, when DR was divided into NPDR and PDR according to its severity , only PDR was significantly associated with incident ASCVD [OR(95%CI): 12.05 (2.63-55.12), P＝0.00].After adjusting for traditional ASCVD risk factors , such an association still existed , with the risk of having ASCVD increasing by 718%[ OR (95% CI): 8.18 ( 1.56 -42.81 ), P ＝0.01] .DR associates strongly with ASCVD in the Chinese population with T 2DM.Conclussion With the severity of DR increasing , the risk of ASCVD also grows.After adjustment for traditional risk factors , PDR is still associated with the risk of prevalent ASCVD.
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Objective To optimize and compare four in vitro cytotoxicity assays in order to find a relatively rapid assay that can replace the traditional 51Cr release assay to evaluate the cytotoxicity of immune cells prepared for immunotherapy.Methods Four assays including BATDA, CAM (calcein acetoxymethyl ester), CytoTox-Glo and PKH were optimized and used to measure the in vitro cytotoxicity of NK-92 cells to K562 cell line.Intra-and inter-assay reproducibility of these assays and their correlation with 51Cr release assay were analyzed.Results After optimization, all of the four cytotoxicity assays showed good correlation with effector to target (E/T) ratio in a certain range.Compared with the other three assays, CytoTox-Glo assay showed obvious hook effect at a high E/T ratio of 40∶1.BATDA assay could detect the significant cytotoxicity of NK-92 cells to K562 cells after incubating them for one hour and that was the shortest time taken by the four assays to detect in vitro cytotoxicity.Both CAM and PKH assays took about four hours and CytoTox-Glo assay took six to eight hours to detect the significant cytotoxic potency.All of the four assays, especially BATDA and CAM assays, showed good intra-and inter-assay reproducibility.Among these assays, BATDA assay showed the best correlation with the traditional 51Cr release assay.BATDA assay, as compared with the other three assays, could be used to detect the cytotoxicity of Caov3 cells, an adherent cell line, and showed good results in evaluating the cytotoxic potency of autologous primary NK cells and CD19-CAR T cells.Conclusion Compared with the other three assays, BATDA assay shows the best linear correlation with 51Cr release assay and has the advantages of time-saving and good reproducibility.Besides, it is a better assay for detecting the cytotoxicity of adherent cells.BATDA assay is a promising substitute for 51Cr release assay in evaluating the in vitro cytotoxic potency of NK cells and other immune cells.
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Objective To establish an assay for the detection of Torque teno sus virus ( TTSuV) strains and to analyze its preliminary application to biologics.Methods Primers and probe were designed according to the conserved sequences.A fluorescent PCR assay for the detection of TTSuV strains was estab-lished.Several parameters including the specificity, linearity, accuracy, sensitivity and anti-interference of the established assay were verified.The fluorescent PCR assay was performed to detect the samples of por-cine blood, cell substrate and rotavirus vaccine.The porcine blood samples positive for TTSuV strains were further genotyped.Results The established fluorescent PCR assay was confirmed to have high specificity as no cross-reactions with parvovirus virus of various species, SV40 and porcine circovirus strains were detec-ted.The linear range of the assay was 1×109-1×103 copies/μl for TTSuV1 genotype and 1×109-1×102 cop-ies/μl for TTSuV2 genotype with a R2 value more than 0.993.The sensitivity of the fluorescent PCR assay was 1×103 copies/μl for TTSuV1 genotype and 1×102 copies/μl for TTSuV2 genotype.The intra-and inter-CVs were both less than 7%in Ct values and less than 25% and 45% respectively in copies.No interfer-ence was found in the detection of TTSuV nucleic acids from cell samples.8 out of 20 porcine blood samples were positive for TTSuV strains, among which one sample was positive for TTSuV1 genotype, four samples were positive for TTSuV2 genotype and the rest were positive for both TTSuV1 and TTSuV2 genotypes.Com-pared with the reference strain, strains genotyped as TTSuV1 and TTSuV2 were respectively shared 98%-99%and 98%homologies in sequences.All of the cell substrate and rotavirus vaccine samples detected by the fluorescent PCR assay were negative for TTSuV strains.Conclusion The fluorescent PCR assay for the detection of TTSuV was established successfully, the application of which would further improve the safety of biologics.
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Objective To measure the macular thickness of natural population in Changping district of Beijing by optical coherence tomography (OCT), and to analyze whether there is an increased thickness in diabetic patients.Methods In a cross-sectional population-based study, OCT was performed on 521 subjects (233 with normal glucose homeostasis, 174 with pre-diabetes and 114 with type 2 diabetes (T2D) according to 2011 Health Examination Survey of Changping.The subjects also received physical examination and laboratory measurements including fasting plasma glucose (FPG), oral glucose tolerance test (OGTT)-2 h plasma glucose and hemoglobin A1c (HbA1c).Results The results showed that central subfield thickness (CST) of the retina of men ((246±22) μm) was significantly greater than that of women ((235±26) μm) (P＜0.001).Meanwhile, no significant difference was found in the CST in subjects of different age, HbAlc and body mass index (BMI) (P＞0.05).Difference in thickness of different glucose groups was not seen in central subfield, inner subfields and outer subfields(P＞0.05).Conclusions Retina CST of men was significantly greater than that of women.No significant difference was found in the CST in subjects of different age, HbA1c and BMI.
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Abstract] Objective To establish an assay for the detection of porcine parvovirus ( PPV) and to verify its application for monitoring cells used for production.Methods A pair of primers and one probe were designed according to the conserved sequence encoding non-structural protein 1 (NS1).Based on the designed primers, a real-time fluorescent quantitative PCR assay for the detection of PPV was developed. Several parameters including the linearity, precision, minimum detection limit and anti-interference of the established assay were evaluated.A stock of PPV strains was prepared by infecting swine testicle ( ST) cells with PPV strains.An assay for the detection of PPV infection was developed by using ST cells as sensitive cells.A combined ST cell infection-PCR test was developed by combining the ST cell infection assay with the real-time fluorescent quantitative PCR assay.The sensitivity of ST cell infection-PCR test was analyzed.The cell samples used for production of biological products were detected by using the established assay.Results The real-time fluorescent quantitative PCR assay was specific for the detection of PPV without cross-reaction to other species of parvovirus virus, SV40 virus and other porcine viruses.The linear range of the assay was 1×109-1×104 copies/μl with a R2 value more than 0.98.The sensitivity of the real-time quantitative PCR assay was 1×104 copies/μl.Both of the intra-and inter-coefficient of variation (CV) were less than 5%in Ct values.The intra-and inter-CV in copies of detection were 5%-15% and 30%-40% respectively.The minimum detection limit of the real-time fluorescent quantitative PCR assay was 1CCID50/ml.The PPV strains were detected in cell samples with no interference.The sensitivity of ST cells infection-PCR test was 0.01CCID50/ml.All of the 22 cell samples were negative for PPV by using the real-time fluorescent quanti-tative PCR assay.Conclusion The real-time fluorescent quantitative PCR and the ST cell infection-PCR test for the detection of PPV in cells were established successfully.The application of the two assays was conducive to further enhance the safety of using cells for production and therapy.
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Objective To conduct extensive quality control tests on Madin-Darby Canine Kidney ( MDCK) cells used for the production of influenza vaccine .Methods Tests for characteristics , extraneous agents, endogenous agents and tumorigenicity were performed on MDCK cells according to Chinese Pharma -copeia Book III .Cell lysate and DNA of MDCK cells were tested for oncogenicity in the light of new interna -tional requirements .Results The MDCK cells extracted from canis were adherent cells with an epithelial morphology, whose average number of chromosome was 80±1.No bacteria, fungi and mycoplasma contami-nation were detected . The detection for extraneous and endogenous virus showed that there was no nonspecific virus causing cytopathic effect , hemadsorption , hemagglutination or animal death .Tests for re-verse transcriptase , bovine viruses and canine viruses were all negative .Each nude mouse was injected with 107 viable cells to observe their tumorigenicity .Twelve weeks after cell injection , no node was found at the injection site and in large organs by gross anatomy .There was no significant difference between test group and negative control group .The test for tumorigenicity of viable cells was negative .Cell lysate and cellular DNA collected from equivalent amount of cells were respectively injected into nude mice , and no node forma-tion was found.There was no significant difference between the test cells and negative controls in pathology indicating that the tested MDCK cells were non-oncogenic .Conclusion It showed the possibility of using MDCK cells for the production of influenza vaccine .
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Objective To study the profiling and authentication of human cell lines in cell bank of our department using short tandem repeat (STR) loci and to analyze the situation of cell contamination and misjudgment. Methods Sixty-one human cells including cells collected and preserved by our cell bank and cells from the other departments were detected by the 16 STR loci-method. To analyze the cross contamina-tion between human cell lines, the results were aligned with profiles published by international cell banks. Isoenzyme detection was employed to authenticate the cell species when the STR signal can not be detected. Results Among the 61 cells, specific profiles were produced by 41 ceils and there was no cross contamina-tion. Thirty-six cells had the completely same STR profiles in 9 STR loci with the same cells preserved by ATCC or JCRB, while 5 cells have different profiling just in the vWA loci. The cells referred above can be recognized as correct cells; Eleven cells (18.0%) were the false cells. Among them, cancer cells of tongue named Tca8113 and cancer cell of liver named HHCC(changed to FHCC98 now) had the same profile with HeLa and HeLa S3 respectively; Two ceils both named HUT-102 have the completely different profiles with ATCC; The signal of 4 cells was not be detected, and all of them were determined as hamster cell lines by u-sing isoenzyme detection. Also 2 cells were identified to be mixed cells. Conclusion The phenomenon of cell misjudgment and cross contamination between cells is serious. Authentication of cell lines correctly, es-pecially for the re-authenticatian of domestic self-established cells, is very important for the guarantee of the reliability and reproducibility for scientific researches.
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Objective To study the application of short tandem repeat (STR) profiling in quality control of human cell lines used for biological production. Methods The methods detecting 9 and 16 STR loci to identify human cell lines by PCR-capiilary electrophoresis were established respectively. Human cell lines, which were derived from many corporations and including diploid cell strains used for virus-vaccine production and 293 cell lines used for gene therapy products, were analyzed and compared by these two methods. Results The STR profiling methods used for authentication of human cell lines were established. Most of human diploid cell strains(20/21 ) used for virus-vaccine production from 13 corporations were iden-tiffed as the intended cells and no cross-contamination was found. However, one MRC-5 cells was identified as a false cell line and one MRC-5 had 3 alleles in D13S317 locus. For 12 strains of 293 cell lines, there were significant differences in STR profiling from different manufactures, which was likely be explained that the sources and gene modifications of these 293 cell lines are not well known and their genes are unstable during passage. Conclusion The STR profiling method has the advantages of high sensitivity and specifici-ty, and can be used for authentication of each of human cell lines for biological production.
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60 minutes. There were no significant differences in the three groups in age, sex ratio, C/T ratio, or left ventricular function. Blood samples for analysis were collected before skin incision and at time intervals up to 6 days postoperatively. Analysis of creatine kinase MB, LDH and cardiac-specific troponin I was used for the detection of myocardial damage. Meantime, the ECG was checked for myocardial infarction. After the reperfusion, myocardial tissue was obtained from the free wall of right ventricle myocardial structure studies. Results: The level of cTnI was increased significantly when the time of myocardial ischemia was prolonged. The changes of CK-MB and LDH were not significant in these three groups. Electron microscopy demonstrated the mitochondria of myocardial cell swelled, the myofilament shortened and the sarcoplasmic reticulum vacuolated in group III. The ECG was almost normal in all groups. Conclusion: The cTnI was an early and highly sensitive biochemical marker of ischemic and reperfusion injury during correction of cardiac defects in children. The concentration of cTnI was correlated ischemia with the degree of so evaluation of the release of cTnI could be used to assess myocardial protection during cardiac operation.
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Objective To summarize our experience on laparoscopic splenectomy with amputation of the secondary spleen pedicle. Methods From September 2006 to May 2007, laparoscopic splenectomy with amputation of the secondary spleen pedicle was performed on 13 patients, including 5 cases of traumatic spleen rupture, 2 idiopathic thrombocytopenic purpura, and 6 hypersplenism. Results All the operations were completed under a laparoscope without using hand-assisted procedures. The intraoperative blood loss was 50-800 ml (mean, 350 ml), and the operation time was 150-300 min (mean, 210 min). No complications occurred during and after the operation. The average postoperative hospital stay was 5-9 d (mean, 7.5 d). The patients were followed up for 1-6 months, during which all the patients had normal platelet count. Conclusions Laparoscopic splenectomy with amputation of the secondary spleen pedicle is a feasible, minimally invasive, safe and low-cost procedure.