ObjectiveTo investigate the predictive value of right atrial myocardial fibrosis in the prognosis of isolated tricuspid regurgitation surgery after left heart valve surgery. MethodsThe patients who underwent tricuspid valvuloplasty by the same operator in Guangdong Provincial People's Hospital from April 2016 to August 2021 due to long-term isolated severe tricuspid regurgitation after left heart valve surgery were included in the study. According to the degree of right atrial myocardial fibrosis, the patients were divided into three groups: a mild group, a moderate group, and a severe group. The clinical data of these patients were compared and analyzed. ResultsA total of 75 patients were enrolled, including 16 males and 59 females with an average age of 57.0±8.4 years. There were 30 patients in the mild group, 29 patients in the moderate group and 16 patients in the severe group. In terms of the preoperative data, there were statistical differences in cardiac function grade, right atrial diameter, tricuspid incompetence area among the three groups (P<0.05). In terms of the postoperative data, there were statistical differences among the three groups in the cardiopulmonary bypass time, mechanical ventilation time, ICU monitoring time, complication rate and mortality (P<0.05). Further pairwise comparison showed that, compared with the mild group, the severe group had longer mechanical ventilation time (P=0.024), longer ICU monitoring time (P=0.003) and higher incidence of postoperative complications (P=0.024), while the moderate group had no statistical difference in all aspects (P>0.05); compared with the moderate group, the severe group had longer ICU monitoring time (P=0.021) and higher incidence of complications (P=0.006). ConclusionThe early outcome of tricuspid valvuloplasty in patients with isolated tricuspid regurgitation after left heart valve surgery with severe right atrial myocardial fibrosis is worse than that in the patients with mild and moderate fibrosis, suggesting that the degree of myocardial fibrosis in the right atrium can be a predictor of the effect of tricuspid regurgitation surgery and a judgement indicator of the surgery timing.
Objective To establish a method to isolate the CD4+CD25+ regulatory T cells (Tregs) and to identify the purity and function of these cells. Methods The peripheral blood (8 mL) were collected from the great saphenous vein of 10 rhesus monkeys (4 females and 6 males, aged 4-5 years, and weighing 5-8 kg). The mononuclear cells were isolated with density gradient centrifugation. CD4+ T cells were separated by the Magnetic cell sorting (MACS) negative selection and MACS positive selection. The cell yield rate, the cell viability, and the cell purity were compared between MACS negative selection and MACS positive selection. In CD4+ MACS negative selection, the anti-biotin MicroBeads and biotin-antibody cocktai in CD4+CD25+ Tregs isolation kit non-human primate were used, and in MACS positive selection, the anti-APC MicroBeads in CD4+CD25+ Tregs isolation kit non-human primate and CD4-APC were used. The CD4+ T cells separated by positive selection were selected to obtain CD4+CD25 Tregs with CD25 MicroBeads. The purity, activity, the FoxP3 level, and the suppressive function to concanavalin A (ConA) activated autologous CD4+CD24- effective T cells (Teffs) of CD4+CD25+ Tregs were detected by flow cytometry. Results After CD4+ T cells were separated by MACS negative selection and MACS positive selection, the cell viabilities were all up to 95%, showing no significant difference (P gt; 0.05). The cell yield rate and purity of CD4+ T cells by positive selection were significantly higher than those of CD4+ T cells by negative selection (P lt; 0.05). CD4+CD25+ Tregs can be successfully isolated by MACS double positive selection. The classifying purity was 76.2% ± 8.6%; the cell activity was 93.3% ± 4.7%; and the level of FoxP3 was 74.2% ± 6.9%. The CD4+CD25+ Tregs had suppressive effect on ConA activated autologous CD4+CD25- Teffs. Conclusion MACS double positive selection can be used to isolate high-purity CD4+CD25+ Tregs from the peripheral blood of rhesus monkeys and the process does not influence the activity of CD4+CD25+ Tregs.
Objective To explore a simple, effective and stable method for the isolation and purification of Kupffer cells from rat liver, enabling further study on the structure and function of these cells in vitro. Methods After laparotomy, a catheter was inserted into the portal vein and secured with artery clamp. Then, the rat liver was perfused and digested with solution Ⅰ and solution Ⅱ containing 0.05% collagenase Ⅳ respectively. The cell suspension was centrifuged with isopycnic sedimentation in a two-step Percoll gradient to harvest Kupffer cells. The isolated Kupffer cells were purified by selective adherence after 30 min of cultivation, and identified by evaluation of phagocytosis of India ink and peroxidase staining with DAB through light and electron microscopy. Results It was verified that the viability of isolated Kupfffer cells was more than 90% through Trypan blue staining. Those Kupffer cells could attach to plastic quickly and phagocytose ink, and had the appearance of “fried eggs” in positive peroxidase staining with a purity of 95%. Under the light microscopy, the appearance of newly isolated Kupffer cells was round with uniform shape and size. After two days of culture, Kupffer cells appeared to distend with irregular or stellate shape. The typical features were observed in the transmission electron micrographs. There were numerous pseudopods and occasional cup-like indentations in the cell membrane of Kupffer cells. The cytoplasm contained numerous types of lysosomes and other phagocytotic vesicles. Conclusion The method for isolating and culturing Kupffer cells in this study is effective and stable, and the biological characters are preserved in the cultured cells.
Objective To find a feasible method that can fast isolate seed cells, keratinocyte stem cell and fibroblasts, for composite tissue engineered skin. Methods The foreskin could be attained from posthectomy, the subcutaneous tissue was removed completely, and the full-thick skin was cut into pieces, 2 mm×2 mm in size, then the pieces were submerged into a centrifuge tube containing collagenase Ⅰ in a oscillator. After 3-hour digestion at 37℃, the dermis was dissolved completely with all the fibroblasts in the digestion solution and the epidermis could be separated easily.With more than 10minute digestion in trypsin at 37℃, the epidermal cells could be harvested. Then flowcytometry and FITCimmunofluorescence for cytokeratin 19 of epidermal cells and FITC-immunofluorescence for vimentin of fibroblast were conducted to identify keratinocyte stem cells in the epidermal cells and fibroblasts in the digestion solution. Moreover, epidermal cells and fibroblasts were cultured in vitro for 7 days to investigate their biological behavior. Results Using collagenase Ⅰ combined with trypsin, epidermal cells andfibroblasts could be isolated at one time within 3 hours. Up to 17% cells demonstrated cytokeratin 19 positive in the epidermal cells, with fibroblast vimentin positive. The amount of fibroblast could be enlarged to more than 100 times within 6 days, but the putative keratinocyte stem cells were difficult for subculture. Conclusion Seed cells for composite tissue engineered skin could be harvested fast at one time, that made it possible to reconstruct composite tissue-engineered skin in vitro.
ObjectiveTo systematically review the incidence of social isolation in Chinese elderly population. MethodsCNKI, WanFang Data, VIP, CBM, PubMed, EMbase, Web of Science and CENTRAL databases were electronically searched to collect studies on the incidence of social isolation in China from inception to May 2022. Two reviewers independently screened literature, extracted data and assessed the risk of bias of the included studies, then, meta-analysis was performed by using Stata 15.1 software. ResultsA total of 20 studies involving 86 111 subjects were included. Meta-analysis results showed that the incidence of social isolation among the elderly in China was 27.54% (95%CI 22.15% to 57.74%). The results of subgroup analysis showed that gender, age, educational level, marital status, self-assessment of health, living style, year of publication, and region surveyed were all influential factors of the incidence of social isolation among the elderly in China. ConclusionCurrent evidence shows that the incidence of social isolation among the elderly in China is relatively high. Due to the limited quality and quantity of the included studies, more high quality studies are needed to verify the above conclusion.
ObjectiveTo compare the clinical efficacy between right visual double lumen tube (VDLT) intubation and right common double lumen tube (DLT) intubation in lung isolation technique. MethodsA total of 57 patients undergoing thoracoscopic surgery with right DLT lung isolation technique in the First People's Hospital of Chenzhou City and West China Hospital from June 2020 to June 2021 were randomly divided into two groups: a DLT group (n=29, 16 males and 13 females, with a mean age of 54.3±13.2 years) and a VDLT group (n=28, 18 males and 10 females, with a mean age of 55.1±13.7 years) at 1 : 1 with random number table generated by the computer. The clinical data of the two groups were compared. ResultsCompared with the DLT group, the catheter positioning time in the VDLT group was statistically shorter (74.9±47.5 s vs. 151.6±88.9 s, P<0.001), the right upper lung occlusion rate (21.4% vs. 51.7%) and the intraoperative re-adjustment catheterization rate (14.3% vs. 48.3%) were lower (P<0.05). The quality of lung collapses immediately after thoracotomy (67.9% vs. 24.1%) and 20 minutes after thoracotomy (100.0% vs. 75.9%) were improved (P<0.05). There was no significant difference in the rate of fiberoptic bronchoscope assistance for positioning, or the incidence of pharynx pain and hoarseness between the two groups (P>0.05). ConclusionCompared with common DLT, VDLT is more efficient, accurate and intuitive in the location of right bronchial intubation.
ObjectiveTo evaluate the effect of disinfection measures in the isolation ward of ICU in severe patients infected with the Omilkerong variant of the new coronavirus, and to optimize the infection control measures. MethodsReferring to the patient's epidemiological and nucleic acid sample data, nucleic acid sampling was performed on the isolated ward environment by surface sampling method, and the nucleic acid Ct value of virus was detected by parallel fluorescence PCR method. The aerosol was collected by a wet-wall cyclone sampler, and the nucleic acid Ct value of the virus was detected by parallel fluorescence PCR. ResultsBefore daily disinfection, SARS-CoV-2 positive samples were found on the surface of the patient's ward and air sampling after the tracheoscopic tracheostomy. No positive samples of SARS-CoV-2 were found on the surface of articles and air in the patient's ward after daily disinfection. ConclusionThe daily disinfection measures of the hospital can achieve the effect of SARS-CoV-2 disinfection, which suggests that the surface of the articles in the ward after invasive operations such as tracheostomy and tracheoscopy, especially within 2 meters of the tracheostomy and the tracheostomy, is the key part of disinfection, which needs to be paid attention to.
【Abstract】ObjectiveTo develop a method of adult porcine pancreatic islet isolation.MethodsThe tails of adult porcine pancreas were perfused through the pancreatic duct with 0.1% cold collagenase(type Ⅺ) and incubated at 38.5 ℃.The digested tissue was dispersed in 4 ℃ Hanks balanced salt solution(HBSS).The tissue suspensions were filtered through a 600 μm mesh.The residual tissue was resuspended in cold HBSS,and put in the Ricordi’s chamber and shaken for 5 minutes,then filtered again.The isolated islets were divided into three groups: control group(n=14),Pefabloc(trypsin inhibitor,n=8) group and FOY(trypsin inhibitor,n=5) group.The collagenase solution of the Pefabloc and FOY group was supplemented with 1.0 mmol/L Pefabloc and FOY respectively. ResultsThe islet yields of the Pefabloc group and FOY group 〔(11 848±3 530) islet/g pancreas and (14 496±3 693) islet/g pancreas〕 were significantly higher than that of the control group 〔(8 505±3 349) islet/g pancreas〕,P＜0.05.The activity of pancreatic protein enzyme in digestive fluid after digestion in control group was higher than the activity of pancreatic duct before injection and Pefabloc group(P＜0.01),which the control group, pancreatic duct before injection and Pefabloc group were (114.7±50.0) BAEEU,(4.0±1.8) BAEEU and (5.5±2.7) BAEEU,respectively.The pancreatic duct before injection and Pefabloc group showed no significant difference in statistics. In control group,when the harvest of islet was more than 8 000/g,the activity of pancreatic protoin enzyme was less than that with the harvest of islet below 8 000/g 〔(78.3±26.7) BAEEU vs (137.5±48.4) BAEEU,P＜0.05〕.Islet after purification in control group,Pefabloc group and FOY group showed good insulin secretion ability for different concentration of glucose.ConclusionA higher porcine pancreatic islet yield can be obtained by this method of pancreatic islet isolation and prophylactic administration of trypsin inhibitors consistently produce excellent islet yields.
Carbapenemase producing Enterobacteriaceae (CPE) has emerged as a significant global public health challenge and placing infected patients at risk of potentially untreatable infections. When resistance to carbapenems occurs, there are often few alternative treatments available. Numerous international guidelines have performed systematic and evidence review to identify new strategies to prevent the entry and spread of CPE in healthcare settings. Several key strategies have been shown to be highly effective. Firstly a new strategy that is proven to be effective is the early identification of the CPE carrier patients through active surveillance cultures. While waiting for the screening results, suspected CPE carriers will be put on preemptive isolation in single room and healthcare worker will at the same time practice contact precautions. The active surveillance culture and prompt preemptive isolation will limit the entry and spread of CPE from getting into hospital. Secondly, it is of utmost importance to incorporate enforcement of the basic infection prevention and control best practices in the hospital including, full compliance to hand hygiene, appropriate use of personal protective equipment, execute antibiotic stewardship program to control abuse of antibiotics, effective environmental cleaning and decontamination, staff education and feedback, as well as surveillance of healthcare-associated infections. Such a holistic approach has been shown to be effective in inhibiting CPE from gaining foothold in the hospital.
Objective To study the effect of soybean trypsin inhibitor (STI) on the rat islets’ yield and function during the process of isolation and purification. Methods The rats were divided into experiment group and control group according to whether STI was put into the collagenase. STI (2.0 mg/ml) was put into the collagenase digestive juice of the experiment group and none to the control group. For both of two groups, islets were isolated by situ perfusion collagenase into the rat pancreas and they were purified by un-continuous gradient centrifugation with Ficoll-400. The quantities of the obtained rat islets before and after purification were recorded, and the morphosis and function of the purified rat islets were tested, then their vivo function were observed after islets plantation. Results After digest and before purification, there was no obvious deviation of the obtained islets quantity between two groups 〔(624±38.2) IEQ vs (586±37.7) IEQ, P＞0.05〕; After purification, there were significant deviation in the islets quantity 〔(408±28.3) IEQ vs (189±27.1) IEQ, P＜0.05〕 and purity quotient 〔（93±2.4）% vs （75±2.1）%, P＜0.05〕. For two groups, there was no obvious deviation of the obtained islets in insulin stimulation and secretion experiment as well as their vivo function experiment. Conclusion The ultimate yield and purity quotient of the rat islets can be obviously improved by using collagenase digestive juice with SIT in situ perfusion on the rat pancreas, and it has no obvious effect on the islets function.