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BACKGROUND: The Rh blood group antigens are encoded by the RHD and RHCE genes, which possess a remarkable degree of polymorphism owing to their high homologous structures. These variants of the RH genes can lead to absence or weak expression of antigens. METHODS: Analysis of RHCE genotyping by Polymerase Chain Reaction (PCR-SSP) method specific to detect c.48G, c.48C, 109 bp insertion of IVS2, c.201A and c.307C and RhCE phenotyping, were conducted in 316 Chinese patients in previous study. One patient with discrepancy typing result was collected for further RhCE serologic typing using microcolumn gel method and tube method in saline using monoclonal antibodies. PacBio sequencing was performed for RHCE, RHD and RHAG complete sequence analysis. 3D molecular models of the protein with the wild-type and mutant residue were generated using the DynaMut web server. The effect of the mutation on the protein function was predicted by PolyPhen-2 software. RESULTS: One male patient of Chinese Han was detected with RHCE*C allele showed by PCR-SSP method but ccEE phenotype. Further PacBio sequencing identified one normal RHCE*cE allele and one RHCE*Ce allele carried a novel c.829G > A (p.Gly277Arg) variant, which the encoded amino acid located in the ninth transmembrane segment of RhCE protein. Crystallisation analysis of 3D molecular models revealed that the substitution at Arg277 leads to the formation of additional hydrogen bonds, including weak hydrogen bonds between multiple atoms. It also results in hydrophobic ion interactions between Arg277 and Ala244. This mutation is predicted to have a damaging effect on protein function. CONCLUSION: One novel RHCE*Ce allele with c.829G > A (p.Gly277Arg) variant was identified to resulting in the absence or weak expression of C and e antigens.
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ObjectiveTo analyze the mental health status and the influencing factors among the residents taking standardized residency training during the outbreak and stable period of COVID-19, so as to provide theoretical basis for their mental health education. MethodsOn February 8 to February 10 and April 11 to April 13, 2020, Symptom Checklist 90 (SCL-90), Beck Srivastava Stress Inventory (BSSI) and Simplified Coping Style Questionnaire (SCSQ) were distributed in online chat groups involving all grades of residents taking standardized residency training in the Second Affiliated Hospital of Kunming Medical University. Through two rounds of questionnaire survey, non-probability sampling method was used to obtain survey samples, and their mental status were analyzed. ResultsA total of 159 valid questionnaires were collected in the first round, and 99 valid questionnaires in the second round. The first survey showed that the total score of SCL-90 was (117.69±37.74) and the detection rate of positive symptoms in SCL-90 was 25.8%, and mainly dominated by obsession, fear and interpersonal sensitivity. In the second survey, the results showed that the total score of SCL-90 was (127.19±51.44), and the main positive symptoms included obsession, depression and interpersonal sensitivity, with a positive detection rate of 30.3 %. The first survey found that the mental health status had significant differences among residents of different grades (χ2=7.46, P<0.05), furthermore, the results indicated that lower grade was a risk factor while non-singleton was the protective factor of mental health status (P<0.05), and SCL-90 total score was positively correlated with score of negative coping styles (r=0.45, P<0.01). The second survey also classified lower grade as risk factor and non-singleton as protective factor (P<0.05), and SCL-90 total score was positively correlated with study stress, economic pressure, interpersonal relationship, clinical practice and negative coping styles (r=0.52, 0.46, 0.55, 0.54, P<0.05 or 0.01). ConclusionResidents under standardized residency training have obvious mental health problems during the outbreak of COVID-19, and the problems become more serious during the stable period of COVID-19.
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Objective@#To examine the relative factors of transmural intestinal necrosis(TIN) during multidisciplinary stepwise management facilitating the decision making in patients with acute superior mesenteric vein thrombosis (ASMVT).@*Methods@#Clinical data of patients with ASMVT admitted to Department of General Surgery, Jinling Hospital from January 2009 to June 2017 were reviewed retrospectively. There were 52 males and 37 females, aging (45.9±12.6) years (range: 20 to 69 years). According to the postoperative pathological results and follow-up, the patients were divided into TIN group (n=31) and non-TIN group(n=58, including 18 cases of intestinal stricture). The related factors were compared between ASMVT patients with TIN and patients without TIN by univariate analysis using t test, U test and χ2 test accordingly, and factors with statistically significance were subsequently submitted to binary Logistic regression analysis. The predictive value and cut-off point of factors were evaluated by receiver operator characteristic(ROC) curve and area under the curve.@*Results@#In univariate analysis, smoking, hypertension, peritonitis, white blood cell count,haemoglobin, international normalized ratio, blood albumin, thrombosis of superior mesenteric branches vein, free intraperitoneal fluid, decrease of bowel wall enhancement and pneumatosis intestinalis were TIN risk factors (all P<0.05). According to the binary Logistic regression analysis, white blood cell count (OR=1.093, 95%CI: 1.010 to 1.182, P=0.027), thrombosis of the superior mesenteric branches vein (OR=11.519, 95%CI: 1.906 to 69.615, P=0.008), pneumatosis intestinalis (OR=11.140, 95%CI: 2.360 to 52.585, P=0.002) were independent relative factors of TIN in patients with AMI, and the area under the ROC curve of the above factors and predictive model was 0.759 (95%CI: 0.647 to 0.871), 0.745 (95%CI: 0.641 to 0.848), 0.737 (95%CI: 0.621 to 0.854), 0.909 (95%CI: 0.847 to 0.971), respectively. The cutoff value of white blood cell count was 18.1×109/L.@*Conclusion@#White blood cell levels, superior mesenteric vein branch thrombosis and pneumatosis intestinalis are independent predictors of TIN in ASMVT.
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Objective To investigate the influential factors of surgical treatment for ischemic enteropathy secondary to portal venous system thrombosis (PVST).Methods Clinical data of 27 patients with ischemic enteropathy secondary to PVST admitted in our department from January 2009 to Jun 2016 were analyzed retrospectively.These patients were divided into two groups according to different surgical procedures:ischemic bowel resection with primary anastomosis or ostomy.Results There were significant differences between the groups in albumin and prealbumin level (t =3.585,4.194,P <0.05).There were also significant differences for BMI and body fat (t =2.325,2.430,P < 0.05).The average time from the onset PVST to ischemic enteropathy was 20 d.Conclusion There should be awareness of ischemic enteropathy.Intestinal resection and anticoagulation therapy is necessary.The choice of surgical procedures depends on the preoperative nutrition level (albumin,prealbumin).
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Objective To explore the diagnosis and treatment of mesenteric venous thrombosis secondary to long-term ischemic enteropathy.Methods The retrospective cross-sectional study was conducted.The clinical data of 36 patients with mesenteric venous thrombosis secondary to long-term ischemic enteropathy who were admitted to the Nanjing General Hospital of Nanjing Military Command from January 2009 to June 2017 were collected.Diagnostic methods:history inquiry,physical examination,laboratory test and image finding.Treatment methods:parenteral nutrition support,selective stage 1 bowel resection with anastomosis or stage 1 bowel resection and colostomy,definitive stage 2 operation for recovering digestive tract.Anticoagulation therapy was performed.Observation indicators:(1) clinical characteristics;(2) treatment;(3) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect postoperative complications up to December 2017.Measurement data with skewed distribution were described as M (P25,P75) and M (range).Results (1) Clinical characteristics:① the main clinical manifestations:34,33,27,20,17 and 14 patients showed respectively discomfort after meal,abdominal pain,abdominal distension,nausea with vomiting,stop of analis exhaust and defecation and weight loss of different degree,and the worst patient lost 20 kg within 1 month.The clinical manifestations of 30 patients were more than 3.② The score of acute physiology and chronic health evaluation (APACHE)Ⅱ in 36 patients was 4 (2,6).③ Laboratory test:total protein (TP),albumin and prealbumin were 55.8 g/L (45.2 g/L,59.1 g/L),30.6 g/L (27.3 g/L,37.5 g/L) and 100.0 g/L (86.0 g/L,132.0 g/L),respectively,showing a decreased trend.④ Imaging finding:enhanced scans of abdominal CT showed the portal cavernous in 16 patients,the absence of main trunk of superior mesenteric vein with extensive collaterals in 12 patients,and dovelopment of portal vein and main trunk of superior mesenteric vein in 8 patients.One patients had intestinal edema and stenosis.X-ray contrast examination of digestive tract showed intestinal stenosis with mucosal erosion in 28 patients and complete intestinal obstruction in 8 patients.(2) Treatment:of 36 patients,24 underwent stage 1 bowel resection with anastomosis and other 12 received stage 1 bowel resection and colostomy (11 undergoing definitive stage 2 operation for recovering digestive tract and 1 refusing stage 2 operation due to advanced age).The length of resected bowel was 30 cm (15 cm,80 cm).One patient with stage 1 bowel resection with anastomosis was complicated with small bowel fistula,and was cured by conservative treatment.There was no complication in other patients.(3) Follow-up:all 36 patients were followed up for 3-10 months,with a median time of 6 months.During the follow-up,7 patients were complicated with secondary portal hypertension,5 of 7 were improved by symptomatic treatment,and 2 died of severe digestive tract bleeding;other patients had no related complications.Conclusions The discomfort after meal and bowel obstruction are the main clinical manifestations of mesenteric venous thrombosis secondary to long-term ischemic enteropathy.The abdominal CT with X-ray contrast examination benefits to diagnosis of patients with poor nutrition status and mild infection.The main strategy includes early surgical resection and postoperative anticoagulation therapy after bleeding risk evaluation.
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Objective@#To evaluate the application of intestinal stomas in mesenteric ischemia (MI) according to the concept of damage control surgery.@*Methods@#Clinical data of 59 MI patients received intestinal stomas at Jinling Hospital, Nanjing University School of Medicine from January 2010 to June 2017 were analyzed retrospectively. There were 41 male and 18 female patients aging of (51±14) years (ranging from 20 to 86 years). All the patients were divided to two groups according to the degree of bowel ischemia: acute MI group (AMI, bowel necrosis, n=43) and chronic MI group (CMI, bowel stricture, n=16). The medium time from onset to consult of AMI was 7(12) days (M(QR)) and the time of CMI was 80(51) days. After the resection of irreversible ischemic intestine, ostomy was carried out for all 59 patients. Patients received oral anticoagulation, enteral nutrition and succus entericus reinfusion therapy for about 6 months after discharge. Then definite surgery to restore digestive tract was preferred.@*Results@#In AMI group, APACHEⅡ score in admission was (16±3). The length of infarcted intestine resected was (160±95) cm, normal bowel left was (220±106) cm. Twelve patients had complications during first post-operation period including sepsis (n=8), acute renal failure (n=4), acute respiratory distress syndrome (n=4), short bowel syndrome (n=4). 30-day mortality was 18.6%. Total 30 patients received operation to restore the continuity of intestinal tract after 202(42) days and APACHEⅡ score was 4±2. Two patients suffered from sepsis and were cured after anti-infection. In CMI group, APACHEⅡ score was 16±3 and NRS2002 score was more than 3. The length of infarcted intestine resected was (43±33) cm. All patients had restored the continuity of intestinal tract after 176 (47) days. No major complications occurred during the first and second post-operation period.@*Conclusions@#According to damage control surgery, after early revascularization, patients with acute intestinal necrosis should be treated with infarcted bowel resection and stomas. Besides, second operation to restore the continuity of intestinal tract should be conducted after nutritional support for 6 months. Patients with ischemic enteropathy who cannot be corrected with severe malnutrition should achieve stomas during first operation.
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Objective To study the molecular mechanism of oridonin-induced apoptosis of ghoma SHG44 cells.Methods A growth curve was plotted using CCK-8 colorimetric method with different concentrations of oridonin (0,1.25,2.5,5,10,20,and 40 μmol/L)to observe its effect on the growth of SHG44 cells.Hoechst33258 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were used to examine the changes in cell morphology and flow cytometry was used to detect cell apoptosis.Western blotting was used to analyze the expression of apoptosis-related proteins (Caspase-3,cleaved Caspase-3,Bax,and Bcl-2)in SHG44 cells.Results SHG44 cell proliferation was significantly suppressed after 24 and 48 h Oridonin treatment,with a half-maximal inhibitory concentration of 7.865 and 4.74 μmol/L,respectively.Hoechst33258 and TUNEL staining showed changes in cell morphology such as shrinkage and nucleus fragmentation and morphogenesis,which are indicative of apoptosis.Western blotting analysis showed that oridonin inhibited the expression of Bcl-2 and activated the expression of Caspase-3 and Bax.Conclusion Oridonin can inhibit the proliferation and induce the apoptosis of SHG44 cells by regulating the expression of apoptosis-related proteins.
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Objective@#To investigate the effect of Gradient treatment for acute superior mesenteric venous thrombosis (ASMVT).@*Methods@#Clinic data of 68 patients of ASMVT admitted in Department of General Surgery, Jinling Hospital, Medical School of Nanjing University from January 2009 to December 2014 were analyzed retrospectively. There were 50 male and 18 female patients with a mean age of (45±12) years. These patients were conducted by the stepwise treatment model (endovascular treatment-damage control surgery-surgical intensive care-intestinal rehabilitation treatment). Clinical outcomes and complications were compared during the follow-up period. Differences about bowel resection length of endovascular treatment and surgical procedures were evaluated with t test.@*Results@#In the 68 cases, 24 cases were cured simply by endovascular treatment, 19 cases received surgical procedures alone (group surgery). Twenty-five patients received endovascular treatment combined with surgical procedures (group combined), including 6 cases temporary abdominal closure. The overall mortality rate was 2.9% (2/68) during hospitalization. The range of bowel resection of group combined significantly reduced compared with group surgery ((92±14) cm vs. (162±27) cm, t=-2.377, P=0.022). During 1-year follow-up period, 4 cases suffered from short bowel syndrome, whom underwent surgery alone.@*Conclusions@#Early diagnosis and treatment is the key to treatment of ASMVT, the rapid improvement of intestinal ischemia is particularly important for prognosis. Combination therapy significantly save more residual small intestine and avoid short bowel syndrome. The selection of early gradient treatment can significantly reduce the mortality and improve the prognosis of ASMVT patients.
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PURPOSE: By using endovascular techniques, we set up an animal model of mesenteric venous thrombosis to avoid surgical laparotomy. MATERIALS AND METHODS: Ten pigs underwent percutaneous transhepatic puncture to create animal model of acute superior mesenteric venous thrombosis. Experimental animals were injected with thrombin via indwelling catheter, while sham-operated animals with receiving physiological saline instead of thrombin. Animals were divided into three groups according to the time of thrombosis: the control (n=3, sham group), group A (n=3, 24h follow up) and group B (n=4, 72 h follow up). Blood samples were collected and tested at the baseline and end of the experiment from the systemic circulation (jugular vein). A pathologist, blinded to the performed interventions, graded the ischemic lesions. RESULTS: Nine pigs were successfully conducted MVT model, while one died of liver rupture during the experiment. White blood cell (WBC) count (group A: 18.77 ± 1.29, group B: 28.93 ± 3.13), D-dimer (group A: 8.30 ± 1.93, group B: 17.30 ± 2.48) and lactate dehydrogenase (LDH) (group A: 452.13 ± 53.14, group B: 753.97 ± 65.29) showed a rapid step-up between the experimental animals and control animals (P<0.05), while hemoglobin (HB) count (group A: 106.63 ± 3.84, group B: 90.00 ± 2.43) and albumin (group A: 35.83 ± 1.15, group B: 27.60 ± 1.44) a steady decline (P<0.05). Changes of these parameters correlated with duration of mesenteric ischemia by statistical analysis (P<0.05). CONCLUSION: The endovascular approach in this minimally invasive porcine model of mesenteric venous thrombosis was feasible. Moreover, further animal studies are underway to evaluate the effectiveness and reproducibility of endovascular technique for MVT model.
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Modelos Animales de Enfermedad , Procedimientos Endovasculares/métodos , Venas Mesentéricas , Trombosis de la Vena/etiología , Animales , Reproducibilidad de los Resultados , PorcinosRESUMEN
Objective To study the effects of ePTFE-covered stents versus bare stents on portal hemodynamics in portal hypertensive patients receiving TIPS. Methods Sixty patients with portal hypertension underwent TIPS with 8 mm diameter bare stents and ePTFE-covered stent from April 2007 to April 2009. The clinical outcomes were observed after TIPS, and the hemodynamics of potal vein system were studied before and after TIPS and during the follow up using direct portography and color Doppler ultrasound technique. Results TIPS procedures were successful in all patients without major complications. The follow-up of patients with bare stents were (8 ± 4) months and follow-up of ePTFEcovered stents group were (6 ± 4) months. Immediately after the TIPS the portosystemic pressure gradient of the two groups decreased by 60% and 58%, respectively(t =0.79, P >0.05). During the follow-up,portosystemic pressure gradient of bare stents group increased gradually, while that in ePTFE-covered stents group maintained low portosystemic pressure gradient (13.2 ± 1.2) mm Hg vs. (9. 5 ± 2. 9) mm Hg, P =0. 015. The blood velocity and volume of venous return of potal vein system were significantly higher in ePTFE-covered stents group than in bare stents group during the follow-up using color Doppler ultrasound method. The blood velocity and volume of the shunts were significantly higher in covered stents group than in bare stents group after 1 year, (125 ±20) cm/s vs. (88 ±13) cm/s, and (1816 ±380) ml/min vs.(1074 ±239) ml/min, respectively P<0. 01. Conclusions In TIPS patients with ePTFE-covered stents high blood velocity and low portosystemic pressure gradient sustained in contrast with those using bare stents.
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Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.