[Identification of rare variants in exons of NLRC4 gene in patients with type 1 diabetes and their impact on gene function].

Objective: To identify rare variants in exon and exon-intron boundary of containing NLR family CARD domain protein 4 (NLRC4) in type 1 diabetes (T1DM) patients, and to explore their effects on gene function. Methods: A total of 508 T1DM patients and 527 healthy controls in the Department of Metabolic Endocrinology, Second Xiangya Hospital of Central South University from August 2017 to September 2020 were selected. The case group included 264 males and 244 females, and the age [M (Q1, Q3)] was [27 (11, 43)] years. The control group included 290 males and 237 females, and their ageï¼»M（Q1，Q3）］was [47 (36, 60)] years old. Identification of rare variants in exons of NLRC4 gene in T1DM patients and healthy controls was performed and verified by next-generation sequencing and sanger sequencing. The NLRC4 gene wild-type and mutant plasmids were constructed and transfected into 293T cells. Western blot (WB) was used to detect the expression of NLRC4 protein and cleavage products of pro-cysteinyl aspartate specific proteinase(procaspase-1). Cycloheximide (CHX) was added to 293T cells transfected with wild-type or mutant NLRC4 plasmid to detect the degradation of NLRC4 protein. The localization of NLRC4 protein was detected by immunofluorescence, and the concentration of IL-1ß in the cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Results: The sequencing results showed that 4 patients and 2 healthy controls had a heterozygous variant c.208C>T in exon 3 of the NLRC4 gene. Two patient had a heterozygous variant c.1564T>C in exon 4, and 1 patients had c.1219G>C in exon 4. These three variants might be pathogenic variants in T1DM. In 293T cells transfected with NLRC4 wild-type and c.208C>T、c.1564T>Cc.1219G>C mutant plasmids, the expression level, degradation rate, localization of NLRC4 protein and the content of cleavage products of procaspase-1 did not change significantly. However, the concentration of IL-1ß secreted by 293T cells transfected with c.1219G>C and c.208C>T plasmid [M(Q1, Q3)] was 15.25 (12.98, 17.52) and 15.44 (13.81, 17.07) ng/L, respectively, which was lower than 18.70 (16.59, 20.81) ng/L of 293T cells transfected wild-type plasmid (P=0.020, 0.010). Conclusions: NLRC4 gene rare variants c.208C>T, c.1564T>C and c.1219G>C may not change the protein expression, degradation and localization, but c.208C>T and c.1219G>C may inhibit the secretion of IL-1ß. This result suggests that NLRC4 rare variants may have an impact on gene function.

[Comparison of clinicopathological features for indications of different types of antiviral therapy in chronic hepatitis B].

Objective: To compare and analyze the clinicopathological features and significance for indications of different types of antiviral therapy in chronic hepatitis B (CHB). Methods: Clinical data of 861 CHB cases who received liver biopsy, had hepatitis B virus (HBV) DNA-positive (> 30 IU/ml) and met the indications for antiviral therapy from January 2014 to December 2019 were included. Liver pathological changes and their correlation with clinical characteristics were compared and analyzed. According to different data, t-test, analysis of variance, nonparametric test, χ2 test, Ridit and logistic regression analysis were used for statistical analysis. Results: Most of the cases (72.24%) had remarkable pathological damage. The degree of liver fibrosis was higher in the normal than the abnormal group (P<0.001). 17.54% cases had hepatic steatosis. The vast majority of cases (97.33%) had positive hepatitis B surface antigen (HBsAg), while only 50.87% had positive hepatitis B core antigen (HBcAg). The positive correlation factors affecting the severity of liver histopathology were alkaline phosphatase level, while the negative correlation factors were positive HBcAg staining, albumin and platelet level. The degree of liver inflammation and fibrosis had statistically significant differences with different HBcAg staining levels (χ2=44.142 and 102.386, respectively; P<0.001), and the severity was more apparent in the negative group. Conclusion: There exist differences in clinicopathological features for indications of different types of antiviral therapy in patients with CHB. Liver function test range is inconsistent with degrees of hepatic histological severity. The positive and intensity of liver tissue HBcAg staining, and albumin and alanine aminotransferase levels have negative correlation with disease severity.

[Clinicopathological analysis in patients with chronic hepatitis B virus infection in immune tolerant phase].

Objective: To analyze the liver pathology, clinical characteristics and influence factors in patients with chronic hepatitis B virus (HBV) infection in immune tolerant phase (IT). Methods: The clinical data of 273 patients in IT phase who underwent liver biopsy from January 2015 to December 2019 were included in this study. The correlation between liver pathological changes and clinical features was analyzed. Results: There were 43 cases (15.75%) with liver histologic activity ≥ G2, 30 cases (10.99%) with liver fibrosis ≥ S2, and 55 cases (20.15%) with liver pathology ≥ G2 and/or ≥ S2. A total of 17.95% patients had liver steatosis. The majority (98.17%) of tissue samples were positive for HBsAg staining, while only 79.49% were positive for HBcAg. The characteristics of liver pathology were comparable in men from women patients. The differences of G and S were not statistically significant according to different HBsAg positivity, while those were statistically significant according to different HBcAg positivity. By univariate and multivariate analysis, the independent risk factors of pathological severity were HBcAg intensity, HBeAg level, and age. However, the differences of liver histologic activity and fibrosis were not statistically significant between those younger than 30 years old group from those older than 30 years old, neither between those younger or older than 40. Although the diagnostic value of liver inflammation and fibrosis 5 (LIF-5) was better than that of aspartate aminotransferase-to-platelet ratio index (APRI) and fibrosis 4 score (FIB-4), three diagnostic models for predicting the pathological severity were not strong enough (all area under the curves<0.8). Only the specificity of LIF-5 for predicting≥ G2, ≥ G2 and/or ≥ S2 was over 80%. Conclusions: Approximately 20% patients with chronic HBV infection in IT phase have progressive liver inflammation or fibrosis. The intensity of liver HBcAg and HBeAg level are negatively correlated with the severity of disease. The diagnostic models or most clinical indicators have low predictive effect for chronic HBV infections in IT phase.

[The establishment and application of testing methods of tetraspanin 7 autoantibody in type 1 diabetes].

Objective: To establish the luciferase immunoprecipitation system assay (LIPS) to test tetraspanin 7 autoantibody (TSPAN7A) and evaluate its value in Chinese type 1 diabetes (T1D) patients. Methods: Renilla luciferase-tagged TSPAN7 plasmids were transfected into 293T cells to obtain Renilla luciferase-tagged TSPAN7 fusion protein. The cell lysate was incubated with sera overnight, followed by addition of protein A-agarose and extensive wash. Finally, the substrate of Renilla luciferase was added and luminescence was detected. Sera from 100 T1D patients [64 males and 36 females,with a mean age of (28±16) years], 119 type 2 diabetes (T2D) patients [78 males and 41 females,with a mean age of (47±12) years] and 98 healthy volunteers [55 males and 43 females,with a mean age of (28±12) years] from the Department of Metabolism and Endocrinology of the Second Xiangya Hospital, Central South University from 2014 to 2017, were tested by LIPS to evaluate the frequency of TSPAN7A. Radioligand binding assay (RLA) was used to test glutamic acid decarboxylase autoantibodies (GADA), protein tyrosine phosphatase-2 autoantibodies (IA-2A) and zinc transporter 8 autoantibodies (ZnT8A). Results: The frequencies of GADA, IA-2A, ZnT8A and TSPAN7A in T1D patients were 72.0%, 40.0%, 29.0% and 25.0%, respectively. After Bonferroni correction, the positivity of TSPAN7A was lower than GADA (P<0.001), but similar with IA-2A (P=0.035) and ZnT8A (P=0.630). The positivity of TSPAN7A in T1D patients was significantly higher than that in T2D (0.84%, P<0.001) and in healthy controls (1.02%, P<0.001). In combination with TSPAN7A, the positivity of islet autoantibodies in T1D patients increased from 82% to 85%. There was no significant difference in clinical characteristics between TSPAN7A-positive T1D and the other three islet autoantibodies-positive patients. Conclusion: This study succeeded in establishing LIPS method to assay TSPAN7A. Moreover, TSPAN7A are valid islet autoantibodies for T1D patients in China.

[Application of ultrasound in the diagnosis and preoperative evaluation of cystic hepatic echinococcosis].

Objective: To investigate the value of ultrasound in the diagnosis and preoperative evaluation of cystic hepatic echinococcosis. Methods: The data of 59 patients (102 lesions) with cystic hepatic echinococcosis confirmed by surgical pathology and serological examination in the Fifth Medical Center of PLA General Hospital from January 2018 to December 2019 were retrospectively analyzed, including 23 males and 36 females; age13 to 64 (42±6) years. The ultrasonographic manifestations were classified, and the involvement of the lesions in the blood vessels and bile ducts was determined. The coincidence rate between ultrasound diagnosis of hepatic echinococcosis and surgical pathology was compared, and the accuracy, sensitivity and specificity of ultrasounic diagnosis of hepatic echinococcosis were analyzed. Results: Ultrasound showed 12 CE1 type lesions, 24 CE2 type lesions, 16 CE3a type lesions, 6 CE3b type lesions, 34 CE4 type lesions, and 10 CE5 type lesions. The coincidence rates of hydatid ultrasound and surgical pathological classification were 91.66% (11/12), 95.80% (23/24), 93.75% (15/16), 83.33% (5/6), 91.17% (31/34) and 100.00% (10/10), respectively. Ultrasound diagnostic efficiency analysis: the accuracy of classification was 94.12%, the sensitivity was 96.94%, and the specificity was 25%. Among the characteristic ultrasound manifestations, the gyrus sign and the polycystic sign were the most frequent, followed by homogeneous thickening of the cyst wall, double wall, separation of the inner cyst wall, and thick wall calcification. There were significant differences between ultrasound in the determination of lesions involving blood vessels and bile ducts and intraoperative findings (all P<0.05). Conclusion: Ultrasound could clearly diagnose cystic hepatic echinococcosis, could better display the imaging characteristics of the disease, and had important clinical significances in indicating the evolution of the disease and determining the involvement of the lesions in the blood vessels and bile ducts.

[Research progress of lipolipomics in primary hepatocellular carcinoma].

Presently, nonalcoholic fatty liver disease has become the most common pathogenic factor of chronic liver disease worldwide that can lead to the occurrence of hepatocellular carcinoma (HCC). Lipid metabolism in cancer cells is closely related to tumorgenesis, invasion and metastasis, and thus acts as one of the hallmark of cancer cells. Lipolipomics is an important branch of metabolomics, which has been adapted recently in the study of HCC for analysis of the structure and function of lipid components by chromatography and mass spectrometry. Fatty acids, glycerides, glycerophospholipids, sphingolipids, and sterol are significantly different in HCC tissues or serum. Therefore, it contributes to the diagnosis, determination of prognosis, mechanistic study and targeted therapy of HCC.

[Postoperative complications and survival analysis of 1 118 cases of open splenectomy and azygoportal disconnection in the treatment of portal hypertension].

Objective: To analyze the recent postoperative and long-term postoperative complications of open-splenectomy and disconnection in patients with portal hypertension. Methods: There were 1 118 cases with portal hypertension who underwent open splenectomy and azygoportal disconnection from April 2010 to September 2015 at Department of Surgery, People's Liberation Army 302 Hospital. Retrospective case investigation and telephone follow-up were conducted in October 2016. All patients had history of upper gastrointestinal bleeding before operation. Short-term complications after surgery were recorded including secondary laparotomy of postoperative abdominal hemostasis, severe infection, intake disorders, liver insufficiency, postoperative portal vein thrombosis and perioperative mortality. Long-term data including postoperative upper gastrointestinal rebleeding, postoperative survival rate and incidence of postoperative malignancy were recorded, too. GraphPad Prism 5 software for data survival analysis and charting. Results: Postoperative short-term complications in 1 118 patients included secondary laparotomy of postoperative abdominal hemostasis(1.8%, 21/1 118), severe infection(2.9%, 32/1 118), intake disorders(1.0%, 11/1 118), liver dysfunction (1.6%, 18/1 118), postoperative portal vein thrombosis(47.1%, 526/1 118)and perioperative mortality(0.5%, 5/1 118). After phone call following-up, 942 patients' long-term data were completed including 1, 3, 5 years postoperative upper gastrointestinal rebleeding rate(4.4%, 12.1%, 17.2%), 1, 3, 5-year postoperative survival rate(97.0%, 93.5%, 90.3%); the incidence of postoperative malignant tumors in 1, 3 and 5 years were 1.7%, 4.4% and 6.2%. Conclusions: Reasonable choosing of surgical indications and timing, proper performing the surgery process, effective conducting perioperative management of portal hypertension are directly related to the patient's short-term prognosis after portal hypertension. Surgical intervention can reduce the rates of patients with upper gastrointestinal rebleeding, improve survival, and do not increase the incidence of malignant tumors.

[Application value of laparoscopic radiofrequency ablation for specific-location hepatocellular carcinoma].

Objective: To investigate the therapeutic efficacy and safety of laparoscopic radiofrequency ablation (LRFA) for specific-location hepatocellular carcinoma. Methods: To retrospectively analyze 496 patients with specific-location hepatocellular carcinoma treated with LRFA from January 2010 to January 2015 in our hospital. There was a total of 652 hepatic lesions with a mean diameter of (2.8±1.3) cm including 397 cases with single lesion and 99 cases with multiple lesions. The hepatic lesions were adjacent to major hepatic vessels, hepatic hilar region, diaphragmatic dome, gallbladder, or gastrointestinal tract and on the surface of the liver, respectively. Results: The 496 patients with 652 hepatic lesions were treated with LRFA successfully.The mean operation time was (48.2±9.6) minutes and the mean LRFA time per lesion was (30.3±8.6) minutes. No severe complications such as bleeding, bile leakage, gastrointestinal tract damage, diaphragmatic injury and liver function failure occurred after operation. The complete necrosis rate of the specific-location hepatocellular carcinomas was 78.4% (389/496) in one month after RFA, partially necrosis rate was 21.6% (107/496) and overall necrosis rate was 100%. In addition, the 1- and 3-year overall survivals (OS) were 95.6% and 88.5%, and progression free survivals (DFS) were 87.9% and 80.8%, respectively. Conclusions: LRFA is a safe, effective, economic and minimally-invasive therapeutic approach for patients with specific-location hepatocelluar carcinoma and has good clinical application value.

[Analysis of the spectrum and resistance of pathogen causing sepsis in patients with severe acute pancreatitis].

Objective: To investigate the characteristics of spectrum and drug resistance of pathogens causing sepsis in patients with severe acute pancreatitis(SAP). Methods: The clinical data of 63 SAP patients with sepsis admitted in Department of Hepatobiliary, People's Liberation Army General Hospital from January 2014 to December 2015 were retrospectively studied. There were 47 males and 16 females, aged from 22 to 73 years, with an average age of (52±11)years. Samples were collected mainly from: (1)pancreatic and peripancreatic necrosis and abdominal drainage; (2)bile; (3) blood or deep venous catheter; (4) sputum and tracheal catheter and thoracic drainage; (5) urine. Strain identification and drug-resistance test were preformed on positive specimens. Results: Of 244 pathogenic isolates, mainly derived from abdominal cavity(36.0%), blood stream (14.0%), central venous catheter(11.8%), necrotic tissue(9.1%) and sputum(8.1%); 154(63.1%) were gram-negative bacteria, 68 cases(27.9%) were gram-positive bacteria and 22 cases(9.0%) were fungi respectively. The top six common pathogens isolated were E. coli(16.0%), E.faecium and faecalis(15.2%), P.aeruginosa(10.7%), K.pneumonia(9.8%), Acinetobacter baumanni(8.2%), Stenotrophomonas maltophilia(5.3%)respectively. The detection rate of E. coli and K. pneumonia extended-spectrum ß-lactamases(ESBL) was 84.6%(33/39) and 70.8%(17/24), the resistance rate to imipeniem was 12.8% and 25.0%, to cefperazone-sulbactam was 28.2% and 29.2%. As to P. aeruginosa and Acinetobacter bacillus, the resistance rate to imipeniem was 50.0% and 75.0%, to cefperazone-sulbactam was 42.3% and 70.0%; Stenotrophomonas maltophilia was completely resistant to cefperazone-sulbactam, but sensitive to minocycline, SMZ-TMP with the resistance rate less than 40.0%. Gram-positive bacterium strains mainly included E. faecium(38.2%, 26/68), E.faecalis(16.2%, 11/68) and Staphylococcus(35.3%, 24/68) which maintained high sensitivity to vancomycin, teicoplanin and linezolid, there was only one isolate resistant to vancomycin. Candida were the sole pathogens of fungal infections, sensitive to common antifungal drugs overall. Conclusions: The gram-negative bacteria are the predominant pathogens mainly including ESBL-producing isolates(E.coli and K. pneumonia) and non-fermentation bacteria(P.aeruginosa and Acinetobacter bacillus) causing sepsis in SAP. The infection rate and drug-resistance rate of these two kinds of pathogens are relatively higher.

[Clinical analysis of 34 cases with sepsis and systemic capillary leak syndrome].

Objective: To study the clinical characteristics of sepsis with systemic capillary leak syndrome(SCLS) and to evaluate the therapeutic effect and clinical significance of fluid therapy adjusted timely in these patients. Methods: The clinical data of 34 patients with sepsis and SCLS in the Department of Hepatobiliary Surgery ICU of General Hospital of People's Liberation Army General Hospital from July 2014 to January 2016 were retrospectively analyzed.There were 21 males and 13 females, aged from 21 to 74 years, with an average age of 56.3 years.Primary disease as follows: 18 cases with severe acute pancreatitis, 7 postoperative cases of subtotal hepatectomy, 5 postoperative cases of pancreatoduodenectomy, 4 postoperative cases of cholelithiasis.These patients were divided into survival group and death group according to their 28-day survival status.The clinical data including C-reactive protein(CRP), platelets (PLT), brain natriuretic peptide (BNP), the level of arterial blood lactic acid(LAC), oxygenation index(PaO2/FiO2, OI), net fluid balance(NFB) and norepinephrine dosage(NE) were collected and compared between two groups at three different intervals(day 1-3, day 4-6, day 7-9). The measurement data and numeration data were statistically analyzed with t test and χ2 test respectively to explore the inherent characteristics of the disease evolution and its clinical significance. Results: The survival group (n=23)and the death group(n=11)had no significant difference in the characteristics of basic clinical characters.The condition of the survival group and the death group were both in progress in 1-3 days period manifested as increased CRP(t=-0.473, P=0.640) and BNP levels(t=0.140, P=0.895), decreased PLT counts(t=-0.505, P=0.620) in the inflammatory response, decreased LAC(t=-1.008, P=0.320) and OI level (t=-2.379, P=0.020)in tissue perfusion index, and positive fluid balance(NFB: t=0.910, P=0.370), required NE(t=-0.853, P=0.400) to maintain effective perfusion pressure with systemic edema in both groups.There was no significant difference of all these clinical parameters between the two groups.The patients' condition of the survival group reached a plateau phase, whereas all relative indicators of the death group implied significant aggravation and deterioration of systemic infection(CRP: t=-3.438, P=0.000; PLT: t=1.649, P=0.110; BNP: t=-10.612, P=0.000), tissue perfusion (LAC: t=-11.305, P=0.000; OI: t=2.743, P=0.010)and tissue edema NFB(t=-4.257, P=0.000) and NE(t=-7.956, P=0.000) in 4-6 days period.In the last 7-9 days period the patients' condition of the survival group took a turn for improvement, yet the condition of the death group continued to deteriorate, refractory septic shock developed and multiple organ dysfunction syndrome followed afterwards inevitably(CRP: t=-10.036, P=0.000; PLT: t=6.061, P=0.000; BNP: t=-10.119, P=0.000; LAC: t=-24.466, P=0.000; OI: t=13.443, P=0.010; NFB: t=-8.345, P=0.000; NE: t=-7.121, P=0.000). Conclusions: The condition of patient with sepsis and SCLS would be improved markedly at the critical turning point around 7-9 days period since the effective systemic treatment began.If the infection does not be significantly constrolled and SCLS still remains in a sustained extravasation period in 7-9 days, the prognosis of these patients may be worse and the mortality may be higher than that of the patients mentioned before.

[Analysis of the influence of gender on the clinicopathological features and prognosis of pancreatic cystic neoplasms].

Objective: To compare the clinicopathological features and prognosis of pancreatic cystic neoplasms (PCN) between male and female patients. Methods: A total of 325 patients with histological confirmed PCN from January 2010 to October 2015 were enrolled. The clinicopathological features, laboratory examination, imaging features, treatment and prognosis were analyzed. Results: Of the 325 patients diagnosed as PCN, 104 were male and 221 were female. The average age of male patients was significantly higher than that of female patients (55.0±12.7 vs 47.9±13.4 years, P<0.001), however, the average tumor size of male patients was smaller than that of female patients (3.9±2.7 vs 4.9±3.0 cm, P=0.004). The location of PCN in male patients was predominantly located at pancreatic head and neck, and majority of male patients were mucinous cystic neoplasms. 1-year, 3-year and 5-year survival rates of patients with PCN were 98.5%, 92.6%, and 87.5%, respectively. 1-year, 3-year and 5-year survival rates of male patients were 97.8%, 84.9%, and 77.8%, however, those of female patients were 98.8%, 96.5%, and 92.2%, respectively. Female patients had better prognosis than male patients (χ2=5.543, P=0.019). Elevated CA19-9 (χ2=3.843, P=0.050), perineuronal invasion (χ2=6.250, P=0.012) and lymph node metastasis (χ2=4.529, P=0.033) were important prognostic factors for malignant mucinous cystic neoplasm. Conclusions: Male patients had low incidence of PCN, and were more common for mucinous cystic neoplasm. The long-term outcome of malignant PCN was poor. Even with complete resection, male patients still had worse prognosis. Close follow-up is recommended especially for male patients.

[Analysis of disease spectrum of biliary surgery among 30 years in a single center].

OBJECTIVE: To explore the changing trend and clinical significance of disease spectrum of biliary surgery among 30 years in PLA General Hospital. METHODS: We retrospectively analyzed the clinical data of 21 189 consecutive patients with biliary diseases in PLA General Hospital between January 1985 and December 2014 according to the inclusion criteria. Taking 5 years as a research group, all patients were divided into 6 groups, and then the 30 years of the disease spectrum of biliary surgery was described. RESULTS: (1) From January 1985 to December 2014, the PLA General Hospital performed 21 189 cases of patients with biliary surgery diseases, including 9 460 cases of male, 11 729 cases of female (female∶male=1.24∶1). The average age was 52.1±13.5 years old (6-98 years). (2) Of the all cases, the most patients was cholelithiasis, accounting for 65.26%, and gallbladder stones accounting for 53.61%. The following disease was cholangiocarcinoma, accounting for 12.79%, and hilar cholangiocarcinoma accounting for 5.95%. (3) From the perspective of disease spectrum changing trend, the proportion of patients with cholelithiasis were decreased gradually, from 74.9% to 65.2%, while the proportion of patients with cholangiocarcinoma showed a gradual upward trend, rising from 6% to 14% (P<0.05); and the proportion of bile duct cystic dilatation and gallbladder carcinoma shared a gradual increase(P<0.05), however, the variation of proportion of gallbladder polyps and biliary stricture were not statistically significant (P>0.05). (4) In patients with cholelithiasis, the proportion of patients with gallbladder stones increased gradually from 71.9% to 82%, while the proportion of patients with bile duct stones decreased gradually from 28.1% to 18%. There was no statistical significance in the proportion of different types of cholangiocarcinoma patients (P>0.05) (40% of the patients with hilar cholangiocarcinoma, 25% of the intrahepatic cholangiocarcinoma, and 35% of the extrahepatic cholangiocarcinoma). CONCLUSION: With the time changing, the disease spectrum of biliary surgery has also changed. The proportion of patients with cholelithiasis, especially the patients with bile duct stones, was declining gradually, and the proportion of patients with cholangiocarcinoma was increasing gradually.

[Significance of three-dimensional reconstruction as a method of preoperative planning of laparoscopic radiofrequency ablation].

OBJECTIVE: To discuss the significance of three-dimensional reconstruction as a method of preoperative planning of laparoscopic radiofrequency ablation(LRFA). METHODS: Thirty-two cases of LRFA admitted from January 2014 to December 2015 in Department of Hepatobiliary Surgery, Chinese People's Liberation Army General Hospital were analyzed(3D-LRFA group). Three-dimensional(3D) reconstruction were taken as a method of preoperative planning in 3D-LRFA group.Other 64 LRFA cases were paired over the same period without three-dimensional reconstruction before the operation (LRFA group). Hepatobiliary system contrast enhanced CT scan of 3D-RFA patients were taken by multi-slice spiral computed tomography(MSCT), and the DICOM data were processed by IQQA(®)-Liver and IQQA(®)-guide to make 3D reconstruction.Using 3D reconstruction model, diameter and scope of tumor were measured, suitable size (length and radiofrequency length) and number of RFA electrode were chosen, scope and effect of radiofrequency were simulated, reasonable needle track(s) was planed, position and angle of laparoscopic ultrasound (LUS) probe was designed and LUS image was simulated.Data of operation and recovery were collected and analyzed. Data between two sets of measurement data were compared with t test or rank sum test, and count data with χ(2) test or Fisher exact probability test.Tumor recurrence rate was analyzed with the Kaplan-Meier survival curve and Log-rank (Mantel-Cox) test. RESULTS: Compared with LRFA group ((216.8±66.2) minutes, (389.1±183.4) s), 3D-LRFA group ((173.3±59.4) minutes, (242.2±90.8) s) has shorter operation time(t=-3.138, P=0.002) and shorter mean puncture time(t=-2.340, P=0.021). There was no significant difference of blood loss(P=0.170), ablation rate (P=0.871) and incidence of complications(P=1.000). Compared with LRFA group ((6.3±3.9)days, (330±102)U/L, (167±64)ng/L), 3D-LRFA group ((4.3±3.1) days, (285±102) U/L, (139±43) ng/L) had shorter post-operative stay(t=-2.527, P=0.016), less post-operation ALT changes (t=-2.038, P=0.048) and post-operative TNF-α changes(t=-2.233, P=0.027). Disease-free survival between two groups was significantly different (χ(2)=4.049, P=0.046). Disease-free survival of 12 months survival rates were 77.6% and 65.7% in 3D-LRFA group and LRFA group, respectively.The median disease-free survival was 16.0 months in LRFA group and over 24.0 months in 3D-LRFA group. CONCLUSIONS: Three-dimensional model of liver reconstruction based on image information is a powerful tool in liver surgery planning.It helps to simulate tumor location and vital tubular structure, make plan for interventional treatment, and therefore mean puncture time and operation time is shortened, influence on liver function is reduced, hospital stay is decreased and DFS is prolonged.

[Posterior debridement combined with atlantoaxial fusion to upper cervical tuberculosis].

Objective: To explore clinical results of posterior debridement combined with atlantoaxial fusion for upper cervical Tuberculosis. Methods: From March 2007 to April 2012, 8 patients with upper cervical Tuberculosis underwent posterior debridement combined with atlantoaxial fusion in our hospital were selected for retrospective analysis. 3 cases were males and 5 females, aged 29-65 (43.5±13.2) years. According to the pedicle destruction, using different screws (pedicle screw or laminar screw) fixation.In the preoperative and final follow-up, Japanese Orthopaedic Association score (JOA) and neck disability index (NDI) were used to evaluate neurological function and calculate improvement rate JOA score. At final follow-up, clinical efficacy was evaluated by Odom's grade. situation of internal fixation, fusion of upper cervical were assessed by imaging examination. During follow-up, complications were documented and analyzed. Results: Postoperatively 12 months, all bony fusion were achieved. Tuberculosis were reached clinical cure in 12-18 months. The JOA score increased from 10.5±2.0 preoperatively to 15.6 ±1.1 in final follow-up(P<0.05), and the NDI decreased from 29.9 ± 6.2 preoperatively to 8.6±1.6 (P<0.05). At last follow-up, according to Odom's standard, excellent were obtained in 6 cases (75.0%), good 1 cases (12.5%) and ordinary 1 case (12.5%). No severe complications was documented during follow-up. Conclusions: The treatment of posterior debridement combine with atlantoaxial fusion, and structure grafting and local anti-Tuberculosis drug using intraoperative, not only could obtain reliable clinical efficacy, completely removal of lesions, but also obtain strong stability, which plays an important role in the treatment of cervical tuberculosis.

[Analysis of 113 cases with mucinous cystic neoplasms in a single institutional study].

OBJECTIVE: To describe the clinicopathological characteristics and prognostic factors of mucinous cystic neoplasm(MCN). METHODS: One hundred and thirteen patients with MCN comfirmed by histological examination admitted in People's Liberation Army General Hospital from January 1994 to June 2015 were analyzed, including 21 male and 92 female patients with mean age of (50.2±14.5) years. Forty-eight patients had incidental discovery of their cystic neoplasm for another reason, 65 patients were symptomatic, and abdominal pain or distension was main clinical symptom. Among 113 patients with MCN, 75 cases were MCN with dysplasia, and 38 cases were MCN with invasive carcinoma. Wilcoxon test and χ(2) test, et al were used to analyzed the data, respectively. RESULTS: MCN was mostly located in body and tail of pancreas.Sex, mean age, clinical symptom, tumor size, and CA19-9 showed significant difference between MCN with dysplasia and MCN with invasive carcinoma (all P<0.05). The presence of a solid component (χ(2)=32.460, P=0.000)and main pancreatic duct dilation(χ(2)= 5.729, P=0.022) were significantly associated with malignancy. Fifty-eight patients with dysplasia were followed up, only one patient occurred tumor recurrence. Thirty-one patients with malignancy were followed up, among which there were 22 patients dead, 1-, 3-, 5-year survival of MCN with malignancy was 76.9%, 56.5%, 36.6%, respectively. Lymphatic metastasis and tumor recurrence were important prognostic factors of MCN with malignancy(both P<0.05). CONCLUSIONS: MCN is most affected by old female with no specific symptom, most tumors are located at the body and tail of the pancreas.MCN with dysplasia have excellent prognosis underwent surgery. Even with complete resection, the long-term survival of MCN with malignancy is not satisfied.

Deep sequencing-based analysis of gene expression in bovine mammary epithelial cells after Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae infection.

The goal of this study was to characterize the transcriptome of primary bovine mammalian epithelial cells (pBMECs) and to identify candidate genes for response and resistance to Staphylococcus aureus (strain S108), Escherichia coli (strain E23), and Klebsiella pneumoniae (strain K96) infection. Using Solexa sequencing, approximately 4.9 million total sequence tags were obtained from each of the three infected libraries and the control library. Gene Ontology (GO) analysis of the S108-infected pBMECs showed differentially expressed genes (DEGs) were significantly involved in metabolic processes. In E23-infected pBMECs, DEGs were predominantly associated with cell death and programmed cell death GO terms, while in K96-infected pBMECs, DEGs were primarily involved in metabolic processes and in utero embryonic development. Analysis of the cluster of orthologous groups of proteins showed that the S108-infected, E23-infected and K96-infected pBMECs were significantly involved in "Translation, ribosomal structure and biogenesis", "General function prediction only" and "Replication, recombination and repair". The transcriptome sequences were also annotated for KEGG orthology, and it was found that DEGs in S108-infected pBMECs were significantly involved in oxidative phosphorylation and Parkinson's disease. The clustered pathway terms of the DEGs of the E23-infected pBMECs were found to involve the NOD-like receptor signaling pathway and oxidative phosphorylation, while those of the K96-infected pBMECs were primarily involved in oxidative phosphorylation and apoptosis. Our results have identified a number of immune-related genes that showed changes in gene expression after bacterial infection, and provided insight into the interactions between pBMECs and the bacteria.

Association of ABCB1, CYP3A4*18B and CYP3A5*3 genotypes with the pharmacokinetics of tacrolimus in healthy Chinese subjects: a population pharmacokinetic analysis.

WHAT IS KNOWN AND OBJECTIVE: Tacrolimus (TAC) is metabolized mainly by the CYP3A subfamily and extruded into the intestine by P-glycoprotein, which is encoded by the ABCB1 gene. Several studies have suggested that the CYP3A5*3 genotype influenced the pharmacokinetics (PK) of TAC. The CYP3A4*18B and CYP3A5*3 alleles are clinically important in Chinese subjects because of their relatively high frequency. The present study aimed at evaluating the effects of ABCB1 (C1236T-G2677T/A-C3435T), CYP3A4*18B and CYP3A5*3 genetic polymorphisms on TAC PK in healthy Chinese subjects. METHODS: Data were obtained from a comparative bioavailability study of oral TAC formulations (n = 22). TAC whole blood concentrations were measured by LC-MS/MS. Genetic polymorphisms were determined using a direct sequencing method. Nonlinear mixed-effects modelling (NONMEM) was performed to assess the effect of genotypes and demographics on TAC PKs. RESULTS AND DISCUSSION: Both CYP3A4*18B and CYP3A5*3 polymorphisms affected the TAC PK, whereas ABCB1 genetic polymorphisms and other demographic characteristics did not. The combined genotypes of CYP3A4*18B and CYP3A5*3 had a greater impact than either genotype alone, and they were estimated to account for 28·4% of the inter-subject variability of apparent clearance (CL/F) by NONMEM. The CL/F in subjects with CYP3A4*1/*1-CYP3A5*3/*3 was 10·3 L/h and was 48·5% in those not carrying CYP3A4*1/*1-CYP3A5*3/*3. WHAT IS NEW AND CONCLUSION: This is the first study to extensively explore the influence of CYP3A4*18B, CYP3A5*3 and ABCB1 genetic polymorphisms on TAC PK in healthy Chinese subjects. The results demonstrated that subjects with a combined genotype of CYP3A4*1/*1-CYP3A5*3/*3 may require lower TAC doses to achieve target concentration levels and further investigation is needed in larger populations to confirm the clinical benefits.