AIM: To examine the changes in immune cells which might be involved in systemic immune dysfunction after severe burns, we detected the sequence of expressed sequence tag in lymphocytes and monocytes of F344 rats post-burn. METHODS: Two pairs of primers were designed depending on the sequence of AI764697.1. SMART RACE technology was employed for amplification of whole sequence. Furthermore, the amplified sequence was testified by Northern blot. RESULTS: A 592bp sequence, which was testified by Northern blot, was suceessfally obtained in our experiments. This product was accepted by GenBank and its accession number is AF244895. CONCLUSION: A cDNA whole sequence in immune cells of F344 rats after severe burns was amplified successfully. More research work should be done to identify the resource, location in genome and function of this product.

Objective To evaluate the feasibility, advantages and pitfalls of trans-gastric, trans-colonic and trans-umbilical peritoneoscopy, and to compare pure natural orifice transluminal endoscopic surgery (NOTES) with hybrid NOTES. Methods Two female pigs of 30kg were used in the study, one for pure NOTES, the other for hybrid NOTES. Each pig received trans-gastric, trans-colonic and trans-umbilical peritoneoscopy, respectively, followed by postmortem examination for organic lesions and closure of incisions.The two approaches were compared in terms of intra-operative situation and manipulation. Results The vital signs of the pigs were stable during the procedures, with no peri-incision organic lesions. There were blind areas in trans-gastric as well as in trans-colonic peritoneoscopy. It was easier to close the colonic incision than the gastric one. The situation in trans-umbilical peritoneoscopy was similar to laparoscopic surgery. Hybrid NOTES seemed superior to pure NOTES with regard to safe access, stable pneumo-peritoneum and adequate visualization. Conclusion Three routes for NOTES appear to be reasonable and feasible. Hybrid NOTES is more effective and safer than pure NOTES with current instruments.

AIM: To investigate the variation of nitric oxide(NO) and NO synthase(NOS) in rats during the early stage of severe burn and their possible relation with prognosis of severe burns.METHODS: Levels of NO - 2/NO - 3, the metabolic products of NO, nNOS and iNOS protein in brain, lung and duodenum of rats were measured before and after burns. Survival times of rats in each group were also measured.RESULTS: Levels of NO - 2/NO - 3 in rats after burn increased remarkably, selective inducible NOS( iNOS) inhibitor aminoguanidine (AG), and nonselective NOS inhibitor L-NAME can inhibit this increasing. Levels of neuronal NOS(nNOS) protein in normal rats were low, and iNOS could not be detected. Levels of nNOS protein increased mildly in all observed tissues and the levels of iNOS protein increased remarkably after burn. Administration of L-NAME and AG made the increase of nNOS more apparently but could not affect the level of iNOS. Survival time of rats decreased in L-NAME group and increased in AG group compared to control group.CONCLUSION: Symptoms such as vascular ralaxation and hypotension in burn shock are connected mainly with over-increased iNOS. [

AIM: To identify the effects of nitric oxide synthase (NOS) inhibitor on NO production, expression of NOS and mean artery pressure (MAP) in rats with severe burns. METHODS: After administration of non-selective NOS inhibitor, L-NAME, and selective inducible NOS (iNOS) inhibitor, aminoguanidine (AG), to rats with severe burns, levels of NO - 2/NO - 3 in blood, mRNA expression of nerve NOS (nNOS) in lung and duodenum, MAP in each group were calculated. RESULTS: Levels of NO - 2/NO - 3 in blood of rats increased significantly post burn, which could be inhibited by L-NAME and AG, especially by L-NAME. Expression of nNOS mRNA in lung and duodenum of rats increased post burn, which could be enhanced by AG and L-NAME. MAP of rats decreased gradually post burn and administration of AG could slow down this process significantly. CONCLUSION: cNOS and iNOS could play different roles in the pathophysiology of burn shock. Over-expression of iNOS could be closely related to the pathogenesis of burn shock.

AIM: To study the effect of cGMP-dependent protein kinase (PKG) on the pathogenesis of burn shock. METHODS: Confluent endothelial cells were disintegrated and centrifugated to obtain cell lysates after being treated with 10% burn serum or PKG activator 8-Br-cGMP. PKG activity of lysates was measured with radioactive isotope label method in a reaction system of phosphorylation of specific substrate H2B by PKG, and the shape and the distribution of intracellular filamentous actin were detected by specific fluorescence staining. For the control study, the PKG specific inhibitor KT5823 were used to pretreat the endothelial cells before the administration of burn serum or PKG activator 8-Br-cGMP. RESULTS: Exposures to burn serum and 8-Br-cGMP led to a rapid time-dependent increase in endothelial PKG activity and the polar distribution of intracellular filamentous actin, and preincubation with KT5823 abolished those effects. CONCLUSIONS: The results suggest that burn serum induces PKG activation and the stress variety of filamentous actin in the vascular endothelial cells, which probably contributes to the endothelial hyperpermeability after burn shock. [

Objective To explore the role of p16 gene methylation in fibroblasts in the occurrence and development of keloid.Methods Skin tissue specimens were resected from the lesions of patients with keloid and normal skin of healthy human controls.Fibroblasts were isolated from these tissue specimens and subjected a primary culture.An immunohistochemical analysis was performed to measure the expression of p16 protein in tissue specimens,real-time fluorescence-based quantitative PCR to determine the mRNA expression level (expressed as 2-△△Ct) of p 16 and DNA methyltransferases (DNMTs) in fibmblasts,and bisulfite sequencing PCR (BSP) to estimate the methylation status of p16 gene in the tissue specimens and primary fibroblasts.Results The keloid fibroblasts (KFbs) showed significandy lower mRNA expression of p16 gene (0.64 ± 0.18 vs.1.92 ± 0.23,t =10.54,P＜ 0.05),but significantly higher mRNA expressions of 3 DNMTs (DNMT1:2.58 ± 0.23 vs.1.13 ± 0.21,t =11.22,P ＜ 0.05; DNMT3A:4.87 ± 0.46 vs.2.38 ± 0.32,t =10.81,P＜ 0.05; DNMT3B:1.57 ± 0.12 vs.0.57 ± 0.16,t =12.45,P＜ 0.05) compared with the normal fibmblasts (NFbs).The DNA methylation rate in the p16 gene promoter region was significantly increased in keloid tissue (1.81％ ± 0.46％) and KFbs (3.15％ ± 0.94％) compared with normal skin tissue (0.90％ ± 0.35％,F =14.23,P＜ 0.01) and NFbs (0.17％ ± 0.29％,F=37.62,P＜ 0.01).Conclusions The methylation and low expression of p16 gene in KFbs may be associated with the uncontrolled growth of keloid,and DNMTs may play a role in the pathogenesis of keloid.

To propose an equipment and technology package for oxygen preparation and supply under hypoxia conditions in order to meet the requirements of populations on the plateau .Different populations were analyzed in terms of their oxygen supply requirements, and the modes, schemes and equipment for oxygen supply were explored accordingly and then applied.Efforts in the past 20 years have resulted in the advances in the equipment for centralized, mobile and portable oxygen preparation and supply, though the equipment for individual use hads to be improved and the diffused oxygen supply scheme also needs to be further evaluated .

Objective:The study seeks to explore the factors influencing the psychological status and sleep quality of medical workers amid the ongoing COVID-19 pandemic, in order to provide data sources and theoretical basis for the development of relevant psychological intervention programs.Methods:Employing the convenience sampling method, general information questionnaire (age, gender, marital status, educational background, job status, etc.), Generalized Anxiety Disorder-7 and Patient Health Questionnaire, epidemic stress index scale, and sleep quality questionnaire were distributed to medical staff between February 18 and April 3, 2020, using the PEM mental health care platform of by ZhongShengKaiXin for medical staff issued. Descriptive, single factor, and correlation analyses, as well as multiple linear regression analysis were used to analyze the data.Results:Overall, 24, 845 questionnaires were collected from 23 provinces, of which 24, 687 were valid, with a recovery rate of 99.36%. The findings showed that the proportion of medical personnel with symptoms of anxiety and depression was 50.58% and 51.37%, respectively; 16.11% had poor or very poor anti-stress ability; and 71.78% reported poor or very poor sleep quality. There was a positive correlation between anxiety, depression, anti-stress ability, and sleep quality ( P<0.05). Anxiety was positively correlated with depression, stress tolerance, and sleep quality( r=0.787, 0.667, and 0.486, all P<0.001); depression was positively correlated with stress tolerance and sleep quality ( r=0.709 and 0.586, both P<0.001); and stress tolerance was positively correlated with sleep quality ( r=0.452, P<0.001). Multiple linear regression analysis results showed that age, gender, marital status, educational background, professional title, job status, and participation influenced the anxiety levels of medical personnel in the backdrop of the pandemic ( P<0.001). Depression levels of medical staff were influenced by gender, educational background, job position, and participation ( P<0.001), while gender, marital status, educational background, job position, and participation influenced the stress tolerance levels ( P<0.001). The sleep quality of medical workers was influenced by age, gender, job position, participation in the fight against the pandemic, and professional title ( P<0.001). Conclusions:Amid the ongoing COVID-19 pandemic, medical staff reported poor mental health status and sleep quality, which can be attributed to diverse factors. The research findings can be useful for assisting medical staff to strengthen their self-cognition, while also providing certain psychological counseling data and theoretical basis for management departments.

Objective To observe the hemodynamic changes of acute after cardiac resynchronization therapy(CRT) interruption in patients with chronic heart failure (CHF),and explore the benefit of hemodynamic of the reverse remodeling and non-reverse remodeling CHF patients during CRT.Methods 46 CHF patients who had undergone implantation of CRT device for more than 6 months were enrolled in the study.The reverse remodeling and non-reverse remodeling were identified by reduction of LV end-systolic volumes ≥ 15％.The two groups underwent echocardiography,and mitral regurgitation area (MRA),velocity time integral of aortic valve (VTI-AV),velocity time integral of mitral valve(VTI-MV),maximum rising rate of left intra-ventricular pressure(+ dp/dtmax),LVFT/T were measured in on and off mode of CRT.Hemodynamic indicators of two groups were compared in CRT on and off modes.The rate of changes of above mentioned parameters ΔMRA,ΔVTI-AV,ΔVTI-MV,Δdp/dt and ΔLVFT/T were calculated and compared in the two groups.Results MRA had significant increasing in both group after 10 minutes intereuption of CRT (P ＜0.01).Two groups had significant worsening of VTI-AV,VTI-AV,+ dp/dtmax and LVFT/T after 10 minutes interruption of CRT as compared to CRT-on mode (P ＜0.05).ΔMRA,ΔVTI-MV,Δdp/dt of reverse remodeling group significantly higher than non-reverse remodeling group(P ＜0.05).But there was no difference in ΔVTI-AV and ΔLVFT/T(P ＞0.05).Conclusions The hemodynamic index become worsening after 10 minutes interruption of CRT in both group of medium-and long-term CRT patients especially in reverse remodeling group,suggesting that patients of both groups have obtained hemodynamic benefits continuously during CRT.Reverse remodeling group get more benefits than non reverse remodeling group.

OBJECTIVEThe purpose of this study was to establish a palatal organ culture method and to investigate the palatogenesis in vitro.

METHODS20 pregnant 14-day mice were killed, embryos were separated ascetically, and palatal shelves were dissected and placed on a modified Trowell's system. All explants were cultured 24 h and 48 h respectively. Finally, all explants were embedded and stained by Hematoxylin and Eosin.

RESULTSAll explants grew healthy. After incubation for 24 h, medial edge epithelium maintained, whereas after 48 h, medial edge epithelium disappeared, bilateral mesenchymal cells contacted, palates fused.

CONCLUSIONThis method provides an effective way for investigating the etiology of cleft palate in vitro.

Animals ; Cleft Palate ; Epithelium ; Female ; In Vitro Techniques ; Mice ; Organ Culture Techniques ; Palate ; cytology ; Pregnancy