Objective To identify early Klebsiella pneumoniae(KP)infection after liver transplantation and its impact on prognosis.Methods Clinical data of 171 liver transplant recipients were retrospectively analyzed,and they were divided into the non-infection(n=52)and infection groups(n=119)according to the bacterial culture results at postoperative 2 weeks.In the infection group,KP was not detected in 86 cases(non-KP infection group),and KP was cultured in 33 cases(KP infection group).Preoperative,intraoperative and postoperative data were statistically compared between the non-infection and infection groups,and between the non-KP infection and KP infection groups.The risk factors of early KP infection after liver transplantation and the influencing factors of long-term survival of the recipients were analyzed.Results Compared with the non-infection group,model for end-stage liver disease(MELD)score and total bilirubin level were higher,the operation time was longer,the length of postoperative intensive care unit(ICU)stay and the length of hospital stay were longer,the amount of intraoperative red blood cell transfusion was higher,the hospitalization expense was higher,the incidence of severe complications was higher,white blood cell count,absolute neutrophil cell count and neutrophil-to-lymphocyte ratio at postoperative 14 and 30 d were higher,absolute lymphocyte count at postoperative 14 d was lower and hemoglobin level at postoperative 30 d was lower in the infection group.The differences were statistically significant(all P＜0.05).Compared with the non-KP infection group,MELD score,total bilirubin level and aspartate aminotransferase(AST)level were higher,the operation time and the length of postoperative ICU stay were longer,the hospitalization expense was higher,the 90-d fatality was higher,the albumin level at postoperative 14 d was lower,and total bilirubin level at postoperative 30 d was higher in the KP infection group.The differences were statistically significant(all P＜0.05).Among 33 recipients with KP infection,16 cases were resistant to carbapenem antibiotics,and 7 of them died within postoperative 90 d.Seventeen cases were intermediate or sensitive to carbapenem antibiotics,and 4 of them died within postoperative 90 d.Preoperative MELD score ≥17 and operation time≥415 min were the independent risk factors for KP infection after liver transplantation(both P＜0.05).The length of postoperative ICU stay ≥44 h and KP infection were the independent risk factors for long-term prognosis of liver transplantation(both P＜0.05).Conclusions KP infection is an independent risk factor for death after liver transplantation.High preoperative MELD score and long operation time are the independent risk factors for early KP infection after liver transplantation.

Objective:To discuss the effect of knockdown of receptor-interacting protein kinase 3(RIP3)on autophagy,pyroptosis,and ferroptosis in the human renal tubular epithelial HK2 cells under hypoxia/reoxygenation(H/R)conditions.Methods:The lentiviral interference vector plasmid shRIP3 and negative control lentiviral interference vector plasmid shNC were transfected into the HK2 cells and the HK cells were divided into shRIP3 group and shNC group,and the normal cultured untransfected HK2 cells were regarded as blank group.After 48 h of transfection,real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to verify the lentiviral transfection efficiencies.The HK2 cells were divided into control group,H/R group,shNC+H/R group,and shRIP3+H/R group.CCK-8 method was used to detect the survival rates of the HK2 cells in various groups;immunofluorescence staining was used to detect the fluorescence intensities of light chain 3(LC3B)and NLR family pyrin domain-containing 3(NLRP3)proteins in the cells in various groups;Western blotting method was used to detect the expression levels of LC3Ⅱ,LC3Ⅰ,Beclin1,Caspase-1,Gasdermin D(GSDMD),interleukin(IL)-1β,and IL-18 proteins in the HK2 cells in various groups;Kits were used to detect the ferri ion(Fe2+)levels in the cells in various groups.Results:Compared with blank group and shNC group,the expression levels of RIP3 mRNA and protein in the HK2 cells in shRIP3 group were decreased(P<0.05).The CCK-8 method results showed that compared with control group,the survival rate of the HK2 cells in H/R group was decreased(P<0.05);compared with H/R group,the survival rate of the HK2 cells in shRIP3+H/R group was increased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of LC3B protein in the HK2 cells in H/R group was decreased(P<0.05),and the fluorescence intensity of NLRP3 protein was increased(P<0.05);compared with H/R group,the fluorescence intensity of LC3B protein in the HK2 cells in shRIP3+H/R group was increased(P<0.05),and the fluorescence intensity of NLRP3 protein was decreased(P<0.05).The Western blotting results showed that compared with control group,the ratio of LC3Ⅱ/LC3Ⅰ and the expression level of Beclin1 protein in the HK2 cells in H/R group were decreased(P<0.05);compared with H/R group,the ratio of LC3Ⅱ/LC3Ⅰ and expression level of Beclin1 protein in the HK2 cells in shRIP3+H/R group were increased(P<0.05);compared with control group,the expression levels of Caspase-1,GSDMD,IL-1β,and IL-18 proteins in the HK2 cells in H/R group were increased(P<0.05);compared with H/R group,the expression levels of Caspase-1,GSDMD,IL-1β,and IL-18 proteins in the HK2 cells in shRIP3+H/R group were decreased(P<0.05).Compared with control group,the expression levels of GPX4 and SLC7A11 proteins in the HK2 cells in H/R group were decreased(P<0.05);compared with H/R group,the expression levels of GPX4 and SLC7A11 proteins in the HK2 cells in shRIP3+H/R group were increased(P<0.05).Compared with control group,the Fe2+level in the HK2 cells in H/R group was increased(P<0.05);compared with H/R group,the Fe2+level in the HK2 cells in shRIP3+H/R group was decreased(P<0.05).Conclusion:Targeted knockdown of RIP3 can induce the autophagy,inhibit the pyroptosis,and reduce the ferroptosis of the human renal tubular epithelial HK2 cells induced by H/R.

Objective To summarize clinical experience of transabdominal pericardial anastomosis of suprahepatic vena cava of the donor and right atrium of the recipient in liver transplantation for Budd-Chiari syndrome (BCS) complicated with liver cancer. Methods Clinical data of a BCS patient complicated with liver cancer undergoing transabdominal pericardial anastomosis of suprahepatic vena cava and right atrium in liver transplantation were retrospectively analyzed. Results The hepatic vein and suprahepatic vena cava were partially occluded in the patient. Liver transplantation was completed by transabdominal pericardial anastomosis of suprahepatic vena cava and right atrium with beating-heart. In addition, due to pathological changes of the recipient's hepatic artery, splenic artery of the recipient was cut off, distal ligation was performed, and the proximal end was reversed and anastomosed with the common hepatic artery of the donor liver, and the reconstruction of hepatic artery was completed. The surgery was successfully performed. At approximately postoperative 1 week, the function of the liver allograft was gradually restored to normal, and no major complications occurred. The patient was discharged at postoperative 25 d. No signs of BCS recurrence was reported after 8-month follow-up. Conclusions It is safe and feasible to treat BCS by liver transplantation with transabdominal pericardial anastomosis of suprahepatic vena cava and right atrium. BCS patients complicated with liver cancer obtain favorable prognosis.

In order to compare the setting of difference coefficients in DRG point payment in different cities in Zhejiang province, the implementation rules of DRG point payment issued by 11 cities in Zhejiang province were comprehensively analyzed. It was found that the difference coefficients in different cities could be divided into three categories, including hospital coefficients alone, hospital coefficients and grade coefficients weighted, and weighted by hospital coefficients, grade coefficients, personal burden levels, case mix indexes, and head-to-time ratio. Its setting differences included four aspects: connotation composition, weight distribution, threshold value, and classification of medical institutions. The authors suggested that the adjustment cycle should be set scientifically to dynamically adjust the difference coefficient, and the scientific setting of the difference coefficient should be promoted through provincial coordination.

Objective:To explore the long-term preservation value and repair effect of normothermic machine perfusion (NMP) on clinically discarded kidneys.Methods:A case of clinical discarded donor kidney was collected, and NMP was carried out in vitro for 9 hours with recovered blood. The dynamic changes of renal appearance, blood gas and biochemistry analysis of perfusate and renal pathology were recorded. Results:In the second to fifth hour of NMP, the appearance of renal was pink and ex vivo normothermic perfusion assessment score (EVNP) was grade Ⅰ. While, the sixth hour and beyond of NMP, the appearance of kidney turned to dark red and EVNP was grade Ⅲ. The renal perfusion blood flow maintained above 150 ml/min in the first 6 hours and decreased significantly after that, and at the end, was only 50 ml/min. During the whole process of perfusion, urine output was maintained at about 100 ml/h. PO 2 remained above 100 mmHg in the first 5 hours of perfusion and from the 6th hour, was lower than 80 mmHg and continued to decline, and was close to 0 at the end of perfusion. The results showed that although the K + concentration changes in blood and urine in the first 5 hours of NMP had a good consistency, the lactic acid level had been rising. In addition, there was no significant change in the histopathology at the fourth hour of perfusion compared with that before zero-point puncture, and the fibrinous thrombus in glomeruli was improved compared with that before perfusion. However, at the sixth hour after perfusion and before the end of perfusion, the pathological changes of renal tissue were significantly worse. There were a large of thrombosis in glomerular blood vessels, renal tubular atrophy and acute tubular necrosis. Conclusions:NMP can realize the evaluation of extended criteria donors before transplantation, and it proves the feasibility and repair potential of NMP in kidney to a certain extent. At the same time, NMP also provides a new way to expand the source of donor kidney and to pre-treat organ in vitro.

Objective: To develop and validate a deep learning model based on fundus photos for the identification of coronary heart disease (CHD) and associated risk factors. Methods: Subjects aged>18 years with complete clinical examination data from 149 hospitals and medical examination centers in China were included in this retrospective study. Two radiologists, who were not aware of the study design, independently evaluated the coronary angiography images of each subject to make CHD diagnosis. A deep learning model using convolutional neural networks (CNN) was used to label the fundus images according to the presence or absence of CHD, and the model was proportionally divided into training and test sets for model training. The prediction performance of the model was evaluated in the test set using monocular and binocular fundus images respectively. Prediction efficacy of the algorithm for cardiovascular risk factors (e.g., age, systolic blood pressure, gender) and coronary events were evaluated by regression analysis using the area under the receiver operating characteristic curve (AUC) and R² correlation coefficient. Results: The study retrospectively collected 51 765 fundus images from 25 222 subjects, including 10 255 patients with CHD, and there were 14 419 male subjects in this cohort. Of these, 46 603 fundus images from 22 701 subjects were included in the training set and 5 162 fundus images from 2 521 subjects were included in the test set. In the test set, the deep learning model could accurately predict patients' age with an R² value of 0.931 (95%CI 0.929-0.933) for monocular photos and 0.938 (95%CI 0.936-0.940) for binocular photos. The AUC values for sex identification from single eye and binocular retinal fundus images were 0.983 (95%CI 0.982-0.984) and 0.988 (95%CI 0.987-0.989), respectively. The AUC value of the model was 0.876 (95%CI 0.874-0.877) with either monocular fundus photographs and AUC value was 0.885 (95%CI 0.884-0.888) with binocular fundus photographs to predict CHD, the sensitivity of the model was 0.894 and specificity was 0.755 with accuracy of 0.714 using binocular fundus photographs for the prediction of CHD. Conclusion: The deep learning model based on fundus photographs performs well in identifying coronary heart disease and assessing related risk factors such as age and sex.
Humans ; Male ; Retrospective Studies ; Deep Learning ; Fundus Oculi ; ROC Curve ; Algorithms ; Risk Factors ; Coronary Disease/diagnostic imaging*

Background: Patients with functional dyspepsia (FD) are accompanied by different degree of psychological stress, and clinicians usually have insufficient quantitative assessment of patients’psychological stress. Aims: To explore the effect of psychological stress assessed by 4-item perceived stress scale (PSS-4) and 10-item perceived stress scale (PSS-10) on dyspepsia symptoms, anxiety, depression, somatization and quality of life in FD patients. Methods: A total of 357 FD patients met Rome IV criteria from March 2021 to March 2022 at Affiliated Hospital of North Sichuan Medical College were recruited. Score of PSS-4, PSS-10, generalized anxiety disorder-7 (GAD-7), patient healthy questionnaire-9 (PHQ-9), adapted patient healthy questionnaire-15 (adapted PHQ-15), dyspepsia symptom severity (DSS), Nepean dyspepsia index-short form (NDI) were performed. Effects of PSS-4, PSS-10 on dyspepsia symptoms, anxiety, depression, somatization and quality of life in FD patients were analyzed. Results: Correlation analysis showed that PSS-4 (r=0.152, P=0.004) and PSS-10 (r=0.194, P=0.000) were correlated with DSS; PSS-4 (r=0.341, P=0.000) and PSS-10 (r=0.389, P=0.000) were correlated with adapted PHQ-15; PSS-4 (r=0.239, P=0.000) and PSS-10 (r=0.327, P=0.000) were correlated with NDI; PSS-4 (r= 0.561, P=0.000) and PSS-10 (r=0.680, P=0.000) were correlated with anxiety; PSS-4 (r=0.449, P=0.000) and PSS-10 (r= 0.524, P=0.000) were correlated with depression. Multiple linear regression analysis showed that psychological stress assessed by PSS-4 (β=0.180, P=0.000), DSS (β=0.390, P=0.000) and FD classification (β=-0.116, P=0.024) were the influencing factors of NDI, and the psychological stress assessed by PSS-10 (β=0.268, P=0.000), DSS (β=0.360, P=0.000) and FD classification (β=-0.116, P=0.021) were the influencing factors of NDI. Conclusions: Psychological stress assessed by PSS-4, PSS-10 have effects on anxiety, depression, somatization, DSS and NDI in FD patients, and PSS-4 is shorter. These results suggest that PSS-4 can be used clinically to assess quickly and initially the impact of psychological stress on FD patients.

Objective To evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.

Objective To investigate the predictive value of baseline neutrophil-to-lymphocyte ratio (NLR) in the curative effect and prognosis of breast cancer patients treated with neoadjuvant chemotherapy.Methods The clinical data of 304 breast cancer patients received neoadjuvant chemotherapy in Beijing Cancer Hospital during 2010 were retrospectively analyzed.The correlations of baseline NLR with clinicopathological parameters of breast cancer patients were analyzed by Wilcox or Kruskal-Wallis tests,and the correlation of NLR with pathologic complete response (pCR) after neoadjuvant chemotherapy was analyzed by the binary logistic regression.The effect of clinic pathological parameters on the disease-free survival(DFS) of the patients was evaluated by the univariable and muhivariable Cox regression models.Results The medians of NLR in pCR + patients (n =62) and pCR-patients (n =242) were 1.76 and 1.72,respectively,and there was no significant difference between themn (P ＞ 0.05).Multivariate analysis indicated that the TNM stage (P ＜ 0.05) and pCR (P ＜ 0.05) were the independent risk factors influencing the prognosis of breast cancer patients.There was no significant correlation between NLR and 5-year DFS (HR=0.842,95％CI=0.566-1.255,P＞0.05).Condusion NLR lacks the predictive value in the curative effect and prognosis of breast cancer patients treated with neoadjuvant chemotherapy,while TNM stage and pCR are the independent risk factors influencing the prognosis of breast cancer patients.

OBJECTIVETo study genetic polymorphisms of the KIR2DS4 gene among ethnic Hans from southern China.

METHODSGenomic DNA was isolated from 306 unrelated individuals and amplified with KIR2DS4-specific PCR primers. KIR2DS4-positive samples were genotyped for the entire coding sequence by sequencing-based typing (SBT). Assignment of allelic genotypes was accomplished by using Assign 3.5 software. For samples with inconclusive SBT results, RT-PCR products covering the entire coding sequence of the KIR2DS4 gene were subjected to cloning and haplotype sequencing.

RESULTSAmong all tested samples, 297 were demonstrated to have carried the KIR2DS4 framework gene. For KIR2DS4-positive samples subjected to SBT for the entire coding sequences, no background was observed with the obtained sequences. Three of the seven identified alleles were of novel types, which were officially named by the KIR subcommittee of the World Health Organization Nomenclature Committee for Factors of HLA System. The observed frequencies for the 7 alleles were KIR2DS4*00101 (78.8%), *003 (10.5%), *004 (16.0%), *010 (23.2%), *017 (0.3%), *00105 (0.3%) and *018 (0.7%), respectively. Allele KIR2DS4*007 was not found. The overall frequency for normal cell-surface expression KIR2DS4 alleles including 2DS4*00101, *017 and *00105 was 79.4%, and that for non cell-surface expression alleles including 2DS4*003, *004, *010 and *018 was 50.4%. The ratio between the two was 1.6:1.

CONCLUSIONThe present study has elucidated the allelic diversity of KIR2DS4 among ethnic Hans from southern China, which may provide valuable data for transplantation as well as studies on KIR-associated disease and evolution.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Gene Frequency ; Genotype ; Genotyping Techniques ; methods ; Haplotypes ; Humans ; Polymorphism, Genetic ; Receptors, KIR ; genetics ; Sequence Analysis, DNA ; methods