BACKGROUND: Previous studies have examined the bulk transcriptome of peripheral blood immune cells in acquired immunodeficiency syndrome patients experiencing immunological non-responsiveness. This study aimed to investigate the characteristics of specific immune cell subtypes in acquired immunodeficiency syndrome patients who exhibit immunological non-responsiveness. METHODS: A single-cell transcriptome sequencing of peripheral blood mononuclear cells obtained from both immunological responders (IRs) (CD4 + T-cell count >500) and immunological non-responders (INRs) (CD4 + T-cell count <300) was conducted. The transcriptomic profiles were used to identify distinct cell subpopulations, marker genes, and differentially expressed genes aiming to uncover potential genetic factors associated with immunological non-responsiveness. RESULTS: Among the cellular subpopulations analyzed, the ratios of monocytes, CD16 + monocytes, and exhausted B cells demonstrated the most substantial differences between INRs and IRs, with fold changes of 39.79, 11.08, and 2.71, respectively. In contrast, the CD4 + T cell ratio was significantly decreased (0.39-fold change) in INRs compared with that in IRs. Similarly, the ratios of natural killer cells and terminal effector CD8 + T cells were also lower (0.37-fold and 0.27-fold, respectively) in the INRs group. In addition to several well-characterized immune cell-specific markers, we identified a set of 181 marker genes that were enriched in biological pathways associated with human immunodeficiency virus (HIV) replication. Notably, ISG15 , IFITM3 , PLSCR1 , HLA-DQB1 , CCL3L1 , and DDX5 , which have been demonstrated to influence HIV replication through their interaction with viral proteins, emerged as significant monocyte marker genes. Furthermore, the differentially expressed genes in natural killer cells were also enriched in biological pathways associated with HIV replication. CONCLUSIONS We generated an atlas of immune cell transcriptomes in HIV-infected IRs and INRs. Host genes associated with HIV replication were identified as markers of, and were found to be differentially expressed in, different types of immune cells.
Humans ; Acquired Immunodeficiency Syndrome ; Transcriptome/genetics* ; HIV ; HIV Infections/genetics* ; Leukocytes, Mononuclear/metabolism* ; CD4-Positive T-Lymphocytes/metabolism* ; Virus Replication ; Membrane Proteins/metabolism* ; RNA-Binding Proteins/metabolism*

Objective:To study the causes of hemorrhage after laparoscopic pancreaticoduodenectomy (LPD) and to develop countermeasures in its prevention.Methods:The clinical data of 215 patients who underwent LPD at the Department of Hepatobiliary and Pancreatic Surgery of Zhejiang Provincial People's Hospital from December 2013 to May 2020 were reviewed. The patients’ clinical data including gender, age, comorbidities and postoperative complications such as bleeding, pancreatic fistula, biliary fistula and intraperitoneal infection were studied, with the aims to analyze the causes, clinical manifestations and treatment results of post-pancreaticoduodenectomy hemorrhage (PPH) after LPD.Results:Of 215 patients, there were 132 males and 83 females, aged (60.7±10.3) years. PPH occurred in 20 patients, incidence rate was 9.30%(20/215). Early hemorrhage was mainly caused by inadequate hemostasis or loosening of vascular clips, while delayed hemorrhage was mainly caused by gastrointestinal fistula with vascular erosion, arterial injury by intraoperative energy instruments or pseudoaneurysms. Among the 20 patients, 6 patients had early hemorrhage and 14 delayed hemorrhage. There was 1 patient with grade A, 10 with grade B and 9 with grade C hemorrhage. Thirteen patients developed pancreatic fistula, 1 biliary fistula, and 2 intraperitoneal infection. One patient responded well to conservative treatment. Hemostasis was successfully achieved by gastroscopy ( n=1) and interventional therapy ( n=7). Eleven patients required laparotomy for hemostasis. In this study, 14 of 20 patients survivied PPH and 6 patients died. The mortality rate was 30% (6 of 20 patients with PPH). Conclusions:Early hemorrhage was caused by inadequate hemostasis or loosening vascular clips, while delayed hemorrhage was related to gastrointestinal fistula with vascular erosion, arterial injury by intraoperative energy instrument or pseudoaneurysm. Careful hemostasis, adequate protection of blood vessels, and accurate anastomosis should be performed in LPD. DSA angiography should be used for arterial hemorrhage which progressed very rapidly. Interventional therapy including embolism and stenting were means to control arterial bleeding in PPH. Decisive surgical exploration when interventional therapy failed was important in reducing the mortality rate of these patients.

Objective:To explore the action mechanism of suppressing expression of mitogen- activated protein kinase 14（MAPK14）to alleviate glutamate excitatory toxicity and its neuronal protection effect.Methods:Lentivirus-mediated MAPK14 interference vector was synthetized by Shanghai Jikai Gene Chemical Technology Co.，Ltd. Astrocytes were obtained from SD rats 48 hours after birth，which were cultured in vitro and transfected by lentivirus-mediated transfection. According to the random number table，the cells were divided into three groups：（1）un-transfected group（normal group）with normal astrocytes and the cells were cultured in regular medium composed of Dulbecco's?modified Eagle's?medium（DMEM）；（2）negative control group with astrocytes transfected by MAPK14 no-loaded interference vector；（3）lentivirus transfected group with astrocytes transfected by MAPK14 interference vector. Seventy-two hours after transfection，astrocytes were co-cultured with neurons for 48 hours，and then they were cultured in a medium containing glutamate for 2 hours. The detection indexes included the optimal multiplicity of infection（MOI）value for astrocytes transfected by lentivirus vector，mRNA levels of MAPK14 and glial glutamate transporter 1（GLT-1）detected by rPCR 72 hours after transfection，protein levels of MAPK14 and GLT-1 detected by Western blot 72 hours after transfection，level of lactate dehydrogenase（LDH）and mortality of neurons measured by spectrophotometry and flow cytometry 2 hours after culturing in the medium with glutamate. Results:（1）The optimal MOI value for lentivirus transfecting astrocytes was 30，and astrocytes grew well after transfection.（2）Seventy-two after transfection，the mRNA level of MAPK14 in lentivirus transfected group（0.005 7±0.000 6）was significantly decreased as compared with un-transfected group（0.013 1±0.001 1）and negative control group（0.013 9±0.001 0）（ P<0.01），the mRNA level of GLT-1 in lentivirus transfected group（0.009 1±0.001 2）was not significantly changed as compared with un-transfected group（0.008 7±0.000 3）and negative control group（0.008 9±0.001 1）（ P>0.05）.（3）Seventy-two hours after transfection，the protein level of MAPK14 in lentivirus transfected group（0.29±0.04）was significantly decreased as compared with non-transfected group（0.61±0.05）and negative control group（0.63±0.01）（ P<0.01），the protein level of GLT-1 in lentivirus transfected group（0.73±0.06）was significantly increased as compared with un-transfected group（0.20±0.03）and negative control group（0.23±0.09）（ P<0.01）.（4）After astrocytes were co-cultured with neurons and subsequently cultured in the medium containing glutamate for 2 hours，the level of LDH in lentivirus transfected group［（109.67±2.40）U/L］was significantly lower than that in un-transfected group［（141.52±3.88）U/L］and negative control group［（141.29±3.61）U/L］（ P<0.01）. The mortality of neurons in lentivirus transfected group［（38.72±0.26）%］was significantly lower than that in un-transfected group［（52.94±1.36）%］and negative control group［（54.30±1.23）%］（ P<0.01）. Conclusions:The transfection with lentivirus-mediated MAPK14 interference vector can increase expression of GLT-1 in astrocytes to increase glutamate re-uptake and relieve the glutamate excitatory toxicity in neurons，which may provide a new experimental basis for future use of astrocyte gene regulation to alleviate neuronal injury caused by glutamate excitatory toxicity after traumatic brain injury.

Objective To investigate the risk factors for short-term prognosis in patients with HBV-related acute-on-chronic liver failure (ACLF). Methods A retrospective analysis was performed for the clinical data of 119 patients with HBV-related ACLF who were admitted to Mengchao Hepatobiliary Hospital of Fujian Medical University from October 2019 to October 2020, and according to their survival status on day 90, they were divided into death group and survival group. The patients were given antiviral therapy with entecavir or tenofovir. Related clinical data were collected, including alanine aminotransferase (ALT), aspartate aminotransferase, cholinesterase (ChE), albumin (Alb), cholesterol, alpha-fetoprotein, and HBV DNA at baseline, as well as the incidence rate of important complications. Model for End-Stage Liver Disease (MELD) score was also calculated. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-squared test was used for comparison of categorical data between two groups; a logistic regression analysis was used to investigate the influencing factors for the 90-day prognosis of patients with HBV-related ACLF and establish a new predictive model; the receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficiency of the new model in predicting the prognosis of HBV-related ACLF. Results Of all patients, 33 died within 90 days, resulting in a mortality rate of 27.7%. There were significant differences between the survival group and the death group in age, ALT, Alb, ChE, MELD score, and incidence rates of hepatic encephalopathy, primary peritonitis, and hepatorenal syndrome (all P < 0.05). The logistic regression analysis showed that baseline hepatic encephalopathy (odds ratio [ OR ]=10.404, 95% confidence interval [ CI ]: 2.522-42.926, P =0.001), serum Alb at baseline ( OR =0.853, 95% CI : 0.764-0.952, P =0.005), and MELD score at baseline ( OR =1.143, 95% CI : 1.036-1.261, P =0.008) were independent predictive factors for the short-term prognosis of patients with HBV-related ACLF. A new predictive model was established based on the combination of these three indices, and the ROC curve analysis showed that this new model had an area under the curve of 0.833, while MELD score had an area under the ROC curve of 0.672. Conclusion As for the evaluation of the 90-day prognosis of patients with HBV-related ACLF, the new prognostic model established based on hepatic encephalopathy, Alb, and MELD score has a better predictive value than MELD score alone.

Objective:To investigate and analyze the energy metabolism characteristics and the correlation between energy metabolism and the risk of secondary bacterial infection in patients with hepatitis B virus-related chronic liver disease (HBV-CLD).Methods:Data of 183 cases admitted to the Mengchao Hepatobiliary Hospital of Fujian Medical University from November 2017 to November 2020 were retrospectively analyzed. 79 cases of chronic hepatitis B, 51 cases of hepatitis B-related liver cirrhosis, and 53 cases of hepatitis B-related liver failure were collected. Among them patients with liver failure and decompensated liver cirrhosis were defined as severe liver disease group. The Quark RMR indirect calorimetry (COSMED Corporation, Italy) was used to exam the patients' energy metabolism condition, and the incidences of secondary bacterial infection of the patients during hospitalization were recorded. Shapiro-Wilk test and normal QQ plot were used to analyze the normal distribution of continuous variable data, which was consistent with the normal distribution and was described by mean ± standard deviation. In addition, if it did not conform to the normal distribution, the median and interquartile distance were used to describe it. Levene’s test was used to test the homogeneity of variance of the data, which was consistent with the normal distribution. The t-test was used to compare the means of the two groups of samples. One-way analysis of variance was used to compare the mean values of the three groups of samples, and then the Tukey's test was used to compare the two groups. If the variance was uneven or did not conform to the normal distribution, the Wilcoxon rank sum test was used to compare the differences between the two groups. Kruskal-Wallis test (H test) was used to compare the differences between the three groups of samples, and then the Dunnett’s test (Z test) was used for comparison between the two groups. Categorical variable data were analyzed using chi-square test. Logistic regression analysis was used to screen independent risk factors, and the criteria for variable inclusion ( P < 0.05). Results:The respiratory entropy (RQ) and non-protein respiratory entropy (npRQ) of the three groups had statistically significant difference ( P < 0.05). Among them, the RQ and npRQ of the chronic hepatitis B group were higher than hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group. There were statistically significant differences in fat oxidation rate (FAT%) and carbohydrate oxidation rate (CHO%) between the three groups ( P < 0.05). Compared with hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group, chronic hepatitis B group ( P < 0.05) had lower FAT% and higher CHO%. There were no statistically significant differences in the measured and predicted resting energy expenditure and protein oxidation rate (PRO%) between the three groups. The incidence of secondary bacterial infection in patients with severe liver disease was 48.39% (45/93). Compared with the non-infected group, the RQ and npRQ values ??of the infected group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Logistic regression analysis showed that glutamyltransferase, cholesterol, and npRQ were independent risk factors for secondary bacterial infections in patients with severe liver disease. Glutamyltransferase elevation, and cholesterol and npRQ depletion had suggested an increased risk of secondary bacterial infection. Subgroup analysis of patients with hepatitis B-related liver failure also showed that compared with non-infected group, RQ value and npRQ value of secondary bacterial infection group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Conclusion:Patients with hepatitis B virus-related chronic liver disease generally have abnormal energy metabolism. Low RQ, npRQ, CHO% and high FAT% are related to the severity of the disease; while npRQ reduction is related to the risk of secondary bacterial infection in patients with severe liver disease, and thus can be used as a clinical prognostic indicator.

Pancreatic duct stone is a rare pancreatic disease in clinic, which is often associated with chronic pancreatitis, and could seriously damage the quality of life of patients, and even induce pancreatic cancer. The diagnosis is mainly based on imaging examination, and the treatment methods are diverse. It is necessary to follow the principle of individualized treatment and treat it as soon as possible. This article reviewed the etiology, mechanism, diagnosis, classification and treatment of the disease.

Objective To explore the safety and feasibility of laparoscopic hepatectomy combined with radiofrequency ablation for primary liver cancer difficult to manage.Methods A retrospective study was conducted to analyze the clinical data of 16 patients who underwent laparoscopic hepatectomy combined with radiofrequency ablation in the Zhejiang Provincial People's Hospital from Apr 2015 to Dec 2017.Results 2 more tumors were found by intraoperative laparoscopic ultrasound.All patients underwent laparoscopic hepatectomy combined with radiofrequency ablation successfully.There were 2 laparoscopic left hepatectomy combined with radiofrequency ablation,2 laparoscopic right hepatectomy combined with radiofrequency ablation,6 laparoscopic left lateral lobectomy combined with radiofrequency ablation,4 laparoscopic right postrior lobectomy combined with radiofrequency ablation and 2 irregular laparoscopic hepatectomy combined with radiofrequency ablation.The intraoperative blood loss ranged from 100-800 ml.The average operative time was (283 ± 112) min.The length of postoperative hospital stay ranged from 5 to 12 days.The tumor-free survival rate after operation is 100％ (16/16) on 6 to 38 months follow up.Conclusion It is safe and feasible to carry out laparoscopic hepatectomy combined with radiofrequency ablation in difficult to manage primary liver cancer.

Objective To study the safety and feasibility of laparoscopy combined with holmium laser in the treatment of chronic pancreatitis complicated with pancreatic ductal stones.Methods To compare the clinical data in patients who underwent laparoscopy combined with holmium laser (10 patients,group A) with those who underwent laparoscopy only (21 patients,group B) at Zhejiang Provincial People' s Hospital from January 2012 to August 2018.The operation time,intraoperative blood loss,intraoperative conversion rate,pancreatic ductal incision length,postoperative pancreatic fistula rate,length of postoperative hospital stay,residual stone rate and relief of postoperative abdominal pain rate of the two groups were documented and analyzed.Results Three of 31 patients were converted to open surgery.The remaining patients in the two groups were discharged home without any perioperative death.Group A and B were significant differences in the pancreatic ductal incision length (5.0±0.8 vs.6.5±1.0) cm,operation time (289.3±51.6 vs.349.5± 34.7) min,and postoperative hospital stay (8.0± 1.2 vs.10.2± 1.6) d between the two groups (P＜0.05).There were no significant differences in the intraoperative conversion to open rate,intraoperative blood loss,postoperative pancreatic fistula rate,residual stone rate and relief of postoperative abdominal pain rate between the two groups (P ＞ 0.05).Conclusions It was safe and feasible to treat chronic pancreatitis complicated with pancreatolithiasis by laparoscopy.Laparoscopy combined with holmium laser had the added advantages of easy access through the pancreaticojejunostomy,shorter operation time,and less intraoperative blood loss.

To determine the value of radiomics in identifying lymph node (LN) metastasis in patients with rectal nonmucinous adenocarcinoma.
 Methods: Imaging data of 91 patients were retrospectively analyzed (61 in the training set and 30 in the test set). A total of 1 301 radiomics features were extracted from high-resolution T2-weighted images of the whole primary tumor. The least absolute shrinkage and selection operator (LASSO) logistic regression was performed to choose the optimal features and construct a radiomics classifier in the training set. Its discrimination performance was compared with that of morphological criteria by receiver operating characteristic (ROC) curve analysis, which was validated in the test set.
 Results: The radiomics classifier combined with five key features was significantly associated with LN metastasis, which distinguished LN metastasis with an area under curve (AUC) at 0.874 (95% CI 0.787 to 0.960) in the training set, and the performance was similar in the test set (AUC 0.878, 95% CI 0.727 to 1.000). The AUCs according to the morphological criteria in the training set and test set were 0.619 (95% CI 0.487 to 0.752) and 0.556 (95% CI 0.355 to 0.756), respectively. Discrimination of the radiomics classifier was superior to that of morphological criteria in both the two datasets (both P <0.05).
 Conclusion: The radiomics classifier provides individualized risk estimation for LN metastasis in rectal nonmucinous adenocarcinoma patients and it has the advantage over the morphological criteria.
Adenocarcinoma ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Rectal Neoplasms ; Retrospective Studies

Objective To study the surgical treatment of xanthogranulomatous cholecystitis (XGC).Methods We retrospectively analyzed the clinical data of 56 patients with XGC who underwent surgical treatment at the Zhejiang Provincial People's Hospital from May 2010 to May 2017.Results The diagnosis of XGC was confirmed by histopathology.On preoperative examination of the 56 patients,42 patients had various degrees of increase in the CA19.9 levels,41 patients (73.2％) had thickened gallbladder walls with continuous mucosal linings on ultrasonography,CT,or MRI,and 18 patients (32.1％) had thickening of gallbladder walls with low density nodules.Gallbladder stones were present in 51 patients (91.1％) and 4 patients (7.2％) presented with Mirizzi syndrome.The 41 patients (73.2％) who were diagnosed as XGC before operation under laparoscopic surgery and 7 patients (17.1％) were converted to open surgery.The remaining 15 patients (26.8％) underwent open operation directly because of uncertainty in the diagnosis.All the patients had frozen section during operation.The postoperative pathological results included 21 localizedtype (37.5％) and 35 diffuse type (62.5％) of XGC.All 56 patients had no long-term complications on followed-up for 0.5～ 6 years.Conclusions XGC is a special kind of chronic cholecystitis.There is difficulty in differentiating from gallbladder cancer before surgery.The diagnosis of XGC mainly depends on ultrasonography,CT or MRI.Cholecystectomy is the treatment for XGC.Laparoscopic surgery is the first line treatment for XGC.