Objective:To investigate the clinical value of indocyanine green(ICG) fluorescence imaging in complex laparoscopic cholecystectomy.Methods:The data of 96 patients with complicated gallbladder stones with cholecystitis and cholecystitis who underwent laparoscopic cholecystectomy(LC) from July 2018 to August 2020 in Hepatobiliary and Pancreatic Surgery of Huangshi Central Hospital of Edong Healthcare Group were retrospectively analyzed. Before operation, the patients were divided into experimental group( n=44) and control group( n=52) according to whether indocyanine green was injected intravenously. Seven hours before operation, the experimental group was injected with 2.5 mg indocyanine green, the experimental group underwent LC under guidance of ICG fluorescence imaging technology. The control group underwent conventional LC. The recognition rate of common bile duct and cystic duct, complete anatomy time of gallbladder triangle, operation time, intraoperative blood loss, bile duct injury and residual stone rat were compared. The measurement data obeying normal distribution was expressed by ( Mean± SD), and the t test was used comparison between groups, and the chi-square test or Fisher exact probability was used comparison between enumeration data. Results:The operation was successfully performed in both groups, In the control group, 1 case was converted to laparotomy, There was no perioperative death. Before the incision of the serosa of the triangle of the gallbladder, In the experimental group, the common bile duct recognition rate was 84.1%(37/44), the recognition rate of cystic duct was 72.7%(32/44). In the control group, the common bile duct recognition rate was 26.9%(14/52), the recognition rate of cystic duct was 28.8% (15/52). There were statistically significant differences in the recognition rate of common bile duct and cystic duct between the two groups ( P< 0.05). In experimental group, the time of complete dissection of gallbladder triangle, the operation time, the intraoperative blood loss were (30.2±8.6) min, (48.2±9.8) min, (16.3±5.2) mL, and (46.7±13.9) min, (65.2±15.4) min, (26.1±11.3) mL in the control group, there were statistically significant difference in the above indicators between experimental group and control group( P<0.05). There was no extrahepatic bile duct injury and residual stones in the experimental group. In the control group, there was 1 case of right posterior hepatic duct injury, 2 cases of common bile duct injury and 1 case of residual gallstone. There was no significant difference in extrahepatic bile duct injury and postoperative stone residual rate between the two groups ( χ2=3.532, P=0.081). Conclusion:ICG fluorescence navigation is helpful for early identification of common bile duct and cystic duct in laparoscopic complex cholecystectomy, which can avoid iatrogenic bile duct injury and has good clinical value.

Objective:To investigate the clinical application and efficacy of laparoscopic splenectomy combined with disconnection in megalosplenia and portal hypertension.Methods:The clinical data of 58 patients with splenomegaly of portal hypertension treated in the Department of Hepatobiliary and Pancreatic Surgery of Huangshi Central Hospital of Eastern Hubei Medical Group from January 2016 to January 2020 were analyzed retrospectively, they were divided into laparoscopy group ( n=34) and laparotomy group ( n=24), Laparoscopic splenectomy combined with devascularization was performed in the laparoscopic group, and open splenectomy combined with devascularization was performed in the open group.The general data, operation time, intraoperative bleeding, postoperative exhaust time, postoperative hospital stay and the incidence of postoperative complications (abdominal bleeding, B/C pancreatic leakage, abdominal infection, etc.) were compared between the two groups. The measurement data obeying normal distribution was expressed by mean±standard deviation ( Mean± SD), and the t test was used comparison between groups, and the chi-square test or Fisher exact probability was used comparison between enumeration data. Results:The surgery was successful in both two groups. 2 cases in the laparoscopic group were converted to laparotomy, There was no death in perioperative period.The operation time of laparoscopy group was (205.3±28.6) min and that of laparotomy group was (156.4±20.7) min, which was significantly longer than that of laparotomy group ( P=0.012). The intraoperative bleeding volume of laparotomy group was (327.2±39.5) mL, which was significantly higher than that of laparoscopy group (246.5±32.3) mL. there was significant difference between the two groups ( P<0.05). The postoperative exhaust time and postoperative hospital stay in the laparoscopic group were (2.6±1.4) d and (9.7±2.3) d, the laparotomy group were (3.8±1.5) d and (12.9±2.7) d respectively. The laparoscopy group was shorter than the laparotomy group. The difference between the two groups was statistically significant ( P<0.05). There were 0 case of abdominal bleeding, 2 cases of B/C pancreatic leakage and 3 cases of abdominal infection in the laparoscopic group, 1 case of abdominal bleeding, 2 cases of B/C pancreatic leakage and 5 cases of abdominal infection in the open group. The incidence of postoperative complications in the laparoscopic group was lower than that in the open group, but there was no significant difference between the two groups( χ2=2.807, P=0.088). Conclusions:Laparoscopic splenectomy combined with devasculation is safe and feasible, with advantages such as little trauma, quick recovery of postoperative intestinal function and short hospital stay, which benefit patients. However, the operation is difficult and requires high technical and psychological quality of surgeons.

Objective:To analyze the effect and mechanism of gadolinium chloride on hepatic ischemia-reperfusion injury (HIRI) in Sprague Dawley (SD) rats.Methods:Thirty six eight weeks special pathogen free SD rats, were included in the project. The body weight ranged from 200 to 250 g. Thirty six rats were randomly divided into sham operation group, model group and gadolinium chloride group with 12 rats/group. Model of ischemia-reperfusion injury was generated in the rats of model group; In the gadolinium chloride group, preoperative intraperitoneal injection of gadolinium chloride was performed before the model of HIRI was established; In the sham operation group, only the abdomen was opened and closed and the hilum was dissected. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected in the three groups. The relative expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1 β (IL-1β) mRNA were detected by Q-PCR. Western blot was used to detect the expression of markers involved in the Toll like receptor 2/myeloid differentiation factor 88 (MyD88) signaling pathway. Immunohistochemistry staining was used to detect the expression of Fas and Fas ligands in hilar bile duct epithelial cells.Results:ALT and AST were (55±8) U/L, (92±22) U/L in sham operation group, lower than those in model group (1 247±62) U/L, (1 117±60) U/L, respectively, and ALT and AST in gadolinium chloride group were (622±50) U/L and (552±41) U/L, lower than those in model group (all P<0.05). Compared with the sham operation group, the relative expression of TNF-α, IL-1 β, IL-6 mRNA in the model group was significantly higher (all P<0.05), but the expression of those markers were higher than gadolinium chloride group (all P<0.05). Gadolinium chloride down-regulated the expression of Toll like receptor 2/MyD88 signaling pathway in rat with liver ischemia-reperfusion. The percentage of Fas protein positive cells in model group was (40.2±3.8)%, and the percentage of Fas ligand positive cells was (36.9±2.9)%, which was higher than those in gadolinium chloride group (29.7±2.3)% and (23.6±2.1)% with statistically significant differences (all P<0.05). Conclusion:Gadolinium chloride can reduce the injury of liver function and inhibit the expression of inflammatory factors in liver tissue of SD rats with hepatic ischemia-reperfusion, which may play a protective role by down regulating the expression of relative protein in Toll like receptor 2/MyD88 signaling pathway.

Objective To investigate the regulation of miR-939-5p on USP22 gene expression and its effect on liver cancer migration and proliferation.Methods The expression of miR-939-5p in hepatoma cell lines (HepG2,MHCC-97H,SMMC-7721,BEL-7404 and Huh7) and normal liver cell line LO2 was detected by realtime PCR (qPCR).The liver cancer cells with the lowest expression were used as experimental subjects,and transfected with miR-939-5p (Experimental group) or miR-NC (Control group).qPCR was used to detect the transfection efficiency of miR-939-5p.Transwell migration assay and MTT proliferation assay were used to detect the migration and proliferation of hepatoma cells after transfected miR-939-5p.Bioinformatics software predicted target genes for miR-939-5p.The dual luciferase reporter gene verified the interaction of miR-939-5p with the target gene.qPCR and Western blotting were used to detect the expression of target genes at mRNA and protein levels after over-expression of miR-939-5p.Measurement data were expressed as (Mean ± SD),and t test was used for comparison between groups.Results The expression of miR-939-5p was significantly lower in hepatoma cell lines than in normal hepatocytes (P ＜0.01),and the expression of miR-939-5p was the lowest in SMMC-7721 cells (P＜0.01).miR-939-5p was efficiently transfected into SMMC-7721 cells [(1.01 ±0.07) vs (20.12 ± 1.27),P ＜0.01].High expression of miR-939-5p inhibited the migration ability (P ＜ 0.01) and proliferative capacity of liver cancer SMMC-7721 cells (P ＜0.05).The USP22 gene may be a target gene of miR-939-5p.The luciferase reporter gene confirmed that miR-939-5p specifically binds to the 3'-UTR of USP22 mRNA (P ＜ 0.01).USP22 expression was decreased at mRNA and protein levels after high expression of miR-939-5p (P ＜ 0.01).Conclusions The expression of miR-939-5p was down-regulated in hepatocellular carcinoma cell lines,and miR-939-5p inhibited the migration and proliferation of liver cancer SMMC-7721 cells.The molecular mechanism was to interfere with the expression of USP22 gene.

Objective To retrospectively analyze and compare the clinical application value of core-needle biopsy (CNB) histology and fine needle aspiration (FNA) cytology in diagnosing malignant thyroid nodules. Methods A total of 134 patients with 137 thyroid nodules (93 malignant nodules and 44 benign nodules) were included in this study. Under ultrasound guidance, successive use of 22 G fine needle and18 G core-needle to puncture each nodule was performed for sampling of thyroid nodule. Surgical findings and pathological manifestations were compared with clinical follow-up results. The success rate of sampling and the diagnostic accuracy, sensitivity as well as specificity for malignant thyroid nodules were compared among FNA, CNB, and CNB/FNA. Results The success rate of puncture sampling with FNA, CNB and FNA/CNB for thyroid nodules was 89.1％, 97.8％ and 100％ respectively. For malignant thyroid nodules, the diagnostic accuracy of FNA, CNB and FNA/CNB was 79.6％, 91.9％ and 96.4％ respectively, the sensitivity was 81.7％, 94.6％ and 97.8％ respectively, and the specificity was 75.0％, 86.4％ and 93.2％ respectively. The success rate of puncture sampling by using CNB or FNA/CNB was significantly higher than that by using FNA (P<0.01), moreover, the diagnostic accuracy and sensitivity for malignant thyroid nodules by using CNB or FNA/CNB was also remarkably higher than those by using FNA (P<0.01) . Conclusion In making diagnosis of malignant thyroid nodules, CNB is accurate, safe and reliable. CNB can be used as a complementary or alternative technique to FNA in clinical practice.

[Summary] The proportions of CD19+CD24 hi CD38 hi regulatory B cells ( Breg ) and CD4+CD25+Treg in peripheral blood of children with type 1 diabetes mellitus ( T1DM) were detected by flow cytometric analysis. The concentration of interleukin 10 (IL-10) protein was measured by ELISA. The mRNA expression levels of Foxp3, cytotoxic T lymphocyte-associated antigen-4 ( CTLA-4 ) , and glucocorticoid-induced tumor necrosis factor receptor ( GITR) in CD4+T cells and IL-10 in CD19+ B cells were evaluated using real-time PCR. The results showed that the proportions of CD19+CD24hiCD38hiBreg in peripheral blood, the mRNA and protein levels of IL-10 in B cells from patients with T1DM were lower than those from healthy controls (P<0. 05). The mRNA expression levels of Treg associated factors such as Foxp3, CTLA-4, and GITR were lower in CD4+ T cells from children with T1DM compared with the controls(P<0. 05). These results suggest that Breg cell deficiency and dysfunction might be one of the important factors causing cellular immune dysfunction in patients with T1DM.

Objective To investigate the effects of polycyclic aromatic hydrocarbons ( PAHs) on regulatory T ( Treg) cells in newborns.Methods Blood samples were taken from the umbilical cord of sev-en newborn babies.CD4+CD25+T cells were isolated and treated with or without 300 nmol/L of phenan-threne for 72 hours in the presence of 100 IU/ml of IL-2.The expression of forkhead box P3 (Foxp3), sig-nal transducer and activator of transcription 3 (STAT3) and STAT5 were analyzed by 8-color flow cytometry. RT-PCR was performed to detect the expression of DNA ( cytosine-5 )-methyltransferase 1 ( DNMT1 ) , DNMT3a, DNMT3b and IL-4 at mRNA level.Pyrosequencing in combination with bisulphite sequencing was used to evaluate the methylation within the promoter and the Treg-specific demethylated region ( TSDR) of Foxp3 locus.Treg cells were cultured for 7 hours with autologous Tresp at Tresp/Treg ratios of 1 ∶1, 2 ∶1, 4 ∶1 and 8 ∶1 and stimulated with anti-CD3/CD28 beads and IL-2 for the evaluation of the immunosuppres-sive activities of Treg cells.Results (1) PAHs inhibited the expression of Foxp3 and the function of Treg cells collected from newborns.(2) PAHs significantly decreased the expression of STAT5 and Foxp3, but increased the expression of STAT3 (P<0.05).(3)PAHs enhanced the methylation of the promoter and the TSDR within Foxp3 gene.(4) The transcription levels of DNMT1, DNMT3a and DNMT3b in PAHs treated group were significantly higher than those of the control group (P<0.05).(5) More IL-4 was secreted by PAHs treated CD4+CD25+T cells, indicating that IL-4 was negatively correlated with STAT5, but positively correlated with STAT3.Conclusion PAHs decreased the number and inhibited the function of Treg cells in newborns.The possible mechanism might be related to the abnormal expression of STAT3 and STAT5 in-duced by IL-4 as well as the methylation within Foxp3 gene.

Objective To investigate the possible factors for differentiation affecting of neonatal regulatory T cells(Treg). Methods Umbilical cord blood was collected from 200 newborns. Treg number was detected by DNA demethylation in the Foxp3 of Treg - cell - specific demethylatedregion(TSDR)based on high resolution melting anal-ysis(HRMA),concentrations of 7,8 - dihydroxy - 9,10 - epoxy - benzo(a)pyrene(BPDE - DNA)adducts and interleukin - 4( IL - 4)in the supernatants of cord blood by enzyme - linked immunosorbent assay( ELISA),and follow - up questionnaires were carried out till 1. 0 - 1. 5 years,for recurrent wheezing or stubborn eczema in infants and related information on parental history of atopic diseases. Results (1)In wheezing group[(0. 48 ± 0. 05)% ]and ec-zema group[(0. 76 ± 0. 05)% ],the number of Tregs was significantly lower compared with that of the asymptomatic group[(1. 14 ± 0. 08)% ](t = 2. 62,2. 83,all P ﹤ 0. 05);the number of Treg in parental history of atopic group was significantly lower than that of the non - atopic group(P ﹤ 0. 05);but the Treg numbers in the non - atopic group was still lower than that of the asymptomatic group(P ﹤ 0. 05).(2)The concentrations of BPDE - DNA adducts in the wheezing group[(236. 30 ± 6. 59)ng/ L]and the eczema group[(173. 40 ± 7. 38)ng/ L]were higher than those of the asymptomatic group[(111. 01 ± 3. 36)ng/ L](t = 10. 35,6. 53,all P ﹤ 0. 05),while BPDE - DNA adduct concen-trations in the atopic group with parental history of wheezing or eczema in infants were lower than those of the non -atopic group(P ﹤ 0. 05).(3)The concentrations of IL - 4 in the wheezing or eczema group in the supernatants of cord blood was higher than the asymptomatic group(P ﹤ 0. 05). Conclusions Neonatal genetic factors and BPDE - DNA adducts could affect Treg differentiation,which are probably the reasons for the formation of allergic diseases.

Objective To compare complications incidence rates of 8-core and 12-core prostate biopsies guided with transrectal ultrasound retrospectively,and to study intervention ways to decrease complications.Methods The data of 260 consecutive patients undergoing first-time transrectal ultrasound-guided biopsy were analyzed.132 patients underwent 8-core biopsy and 128 patients underwent 12-core biopsy.Results In 8-core group,there were 7 cases of infections,15 cases of gross hematuria,7 cases of rectal bleeding and 6 cases of acute urinary retention,and in 12-core group,5 cases of infections,19 cases of gross hematuria,9 cases of rectal bleeding and 7 cases of acute urinary retention in 12-core group.There are no statistical differences of complications incidence rate between two groups (P ＞ 0.05).when prostate volume ＞ 45 ml,detection rate of prostate cancer in 12-core group was significantly higher than that in 8-core group.Conclusions There are no statistical differences of complications incidence rate between 8-and 12-core prostate biopsy.Homologous measure must be taken to reduce complications.12-core prostate biopsy is recommended as prostate volume ＞ 45 ml.

BACKGROUND:Whether determination of tacrolimus blood concentration by different immunoassay methods can influence predictive ability to immunosuppressive effects and toxicity, and whether it can be more sensitive to reflect blood concentration in patients with renal dysfunction are worthy of studying.
OBJECTIVE:To analyze the correlation of tacrolimus (FK506) concentrations determined by enzyme-multiplied immunoassay technique (EMIT) and enzyme linked immunosorbent assay (ELISA) in combination with renal function parameters.
METHODS:133 clinical blood samples were col ected. EMIT and ELISA techniques were used to determine the FK506 concentration. The correlation of two determination methods were analyzed, combined with renal function. RESULTS AND CONCLUSION:In patients with renal dysfunction, the mean results and standard deviation mensurated by ELISA were higher than those by EMIT. For blood concentration in 5-20μg/L by ELISA, the incidence of renal dysfunction occurred less than by EMIT. The overal mean results of blood concentration for two methods appeared no significant difference (r=0.904 5, P>0.05). When the concentration was less than 2.0μg/L, the concentration results by EMIT were higher than those by ELISA (P<0.01). When the concentration was more than 2.0μg/L, there was no significant difference between two determination methods (P>0.05). These findings indicate that EMIT and ELISA has good correlation, which are both suitable for clinical routine determination of plasma concentration. It is not recommended for applying EMIT method to determine low blood concentrations (<2.0μg/L). The reference range of concentration should be compartmentalized depending on combination of determination methods and renal function.