Primary open angle glaucoma(POAG)is the most common type of glaucoma, with common causes including variations in trabecular tissue and increased venous pressure. POAG has a certain genetic tendency, and POAG with an autosomal dominant inheritance pattern is mainly caused by single gene mutations. Studies have found that the pathogenesis of POAG may be related to key pathogenic genes(MYOC, OPTN, WDR 36), as well as mitochondrial dysfunction, oxidative stress, and epigenetic regulation. At present, the clinical treatment options for POAG mainly include enhancing trabecular meshwork function, inhibiting aqueous humor production, neuroprotection and regeneration of retinal ganglion cells, and applying gene editing technology, all of which have achieved certain results. However, there is no unified research on the relationship between the occurrence of POAG and genes, as well as treatment plans. The article explores new targets and treatment strategies for POAG gene therapy by analyzing the role of genes in the pathogenesis of POAG, aiming to provide reference for clinical treatment of this disease.

Objective:To explore the correlation between abnormal expression of serum 25(OH)D and immune-related fac-tors and subclinical atherosclerosis(AS)in patients with impaired glucose regulation.Methods:A total of 142 patients with im-paired glucose regulation admitted to Zaozhuang Municipal Hospital from December 2019 to April 2021 were selected.The patients with impaired glucose regulation were selected as the control group(n=86)and patients with subclinical atherosclerosis were selected as the observation group(n=56)according to carotid ultrasound examination and bramolecular pulse wave conduction velocity.The serum 25(OH)D and immune factors were compared between the two groups.Carotid ultrasound was used to measure the carotid inti-media thickness,and Pearson method was used to determine the correlation between the intimedia thickness,25(OH)D and immune factors.Baseline data and hematological indicators were collected,univariate and multivariate Logistic regression analysis was per-formed to determine the influencing factors of subclinical atherosclerosis,and ROC curve was used to evaluate the diagnostic efficacy of serum 25(OH)D and immune factors in subclinical atherosclerosis.Results:The serum 25(OH)D[(24.01±4.87)mmol/L vs(30.74±5.01)mmol/L,t=7.909,P=0.000)in the observation group was significantly lower than that of control group.TNF-α[(48.32±8.02)ng/L vs(33.21±9.00)ng/L,t=10.199,P=0.000)and IL-6[(41.22±9.43)ng/L vs(30.21±7.01)ng/L,t=7.492,P=0.000)in ob-servation group were significantly higher than those in control group.Carotid intima-media thickness was negatively correlated with se-rum 25(OH)D(r=-0.428,P<0.001),and was positively correlated with serum TNF-α and IL-6(r=0.574,0.577,P<0.001).Logis-tic regression analysis showed that serum 25(OH)D(OR=0.520,95%CI:0.401～0.675),serum TNF-α(OR=1.667,95%CI:1.131～2.457)and serum IL-6(OR=1.478,95%CI:1.213～1.802)were the influencing factors of subclinical atherosclerosis.ROC curve showed that the optimal cut-off value of serum 25(OH)D was 28.32 mmol/L,and the corresponding sensitivity,specificity and AUC were 69.64%,70.93%and 0.803(95%CI:0.749～0.855),respectively.The critical value of serum TNF-α was 40.56 ng/L,corre-sponding sensitivity was 71.43%,specificity was 72.09%,and AUC was 0.761(95%CI:0.717～0.823).Serum IL-6 cut-off value was 36.13 ng/L,corresponding sensitivity was 60.71%,specificity was 60.47%,and AUC was 0.627(95%CI:0.566～0.702).The sensitivity,specificity and AUC of regression analysis were 85.71%,81.40%and 0.889(95%CI:0.830～0.915).Conclusion:Serum 25(OH)D and immune-related factors alone and in combination can effectively predict the occurrence of subclinical atherosclerosis,which is correlated with carotid intima-media thickness,and serum 25(OH)D and immune-related factors are predictors of subclinical atherosclerosis.

Objective:To evaluate the predictive value of neutrophil free fatty acid receptor 3 (FFAR3) for secondary infection in patients with severe acute pancreatitis (SAP).Methods:① Biological information analysis: peripheral blood microarray data sets related to acute pancreatitis (GSE194331) were obtained from the Gene Expression Omnibus (GEO), including data from 32 healthy adults, 52 patients with mild acute pancreatitis, 20 patients with moderate-to-severe acute pancreatitis, and 10 patients with SAP. The original data of GSE194331 dataset were downloaded for quality control, pruning, quantification, annotation and difference analysis, and the different genes were obtained. ② Clinical study: a prospective observational study was conducted. Forty-five SAP patients admitted to the critical care medicine department of the Eastern Theater Command General Hospital of the Chinese People's Liberation Army from January to November 2022 were enrolled, and they were divided into infected group and non-infected group according to whether secondary infection occurred during intensive care unit (ICU) stay. At the same time, 10 healthy adult volunteers were enrolled as control. Peripheral blood of subjects in each group was collected, neutrophils were isolated, and FFAR3 mRNA expression was detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR). Spearman correlation method was used to analyze the correlation between neutrophil FFAR3 mRNA expression and secondary infection in SAP patients. Multivariate Logistic regression analysis was used to evaluate whether neutrophil FFAR3 mRNA expression was a risk factor for secondary infection in SAP patients. Receiver operator characteristic curve (ROC curve) was plotted to evaluate the predictive value of neutrophil FFAR3 mRNA expression on secondary infection in SAP patients.Results:① Results of biological information analysis: the analysis of GSE194331 dataset showed that 301 genes were differentially expressed in peripheral blood cells between healthy controls and patients with pancreatitis. By biological function analysis, 8 biological functions involved in immune response were obtained, and 44 differential expressed genes were enriched in these 8 biological functions. The results of cell distribution analysis showed that there were 21 differential expressed genes expressions on neutrophils significantly higher than other immune cells, and the gene related to lipid metabolism was FFAR3. These results indicated that FFAR3 expression was closely related to the occurrence and development of SAP. ② Clinical study results: out of the 45 SAP patients, 24 developed into secondary infection during ICU stay, 21 did not develop into secondary infection. The expression of neutrophil FFAR3 mRNA in SAP patients with secondary infection was significantly higher than that in SAP patients without secondary infection and healthy controls [2 -ΔΔCt: 3.8 (3.0, 4.2) vs. 1.4 (1.1, 2.7), 1.0 (0.8, 1.1), both P < 0.05]. Spearman correlation analysis showed that neutrophil FFAR3 mRNA expression was positively correlated with secondary infection in SAP patients ( r = 0.799, P < 0.001). Multivariate Logistic regression analysis showed that increased FFAR3 mRNA expression was an independent risk factor for secondary infection in SAP patients [odds ratio ( OR) = 17.212, 95% confidence interval (95% CI) was 3.004-98.613, P = 0.001]. ROC curve analysis showed that the area under the ROC curve (AUC) of neutrophil FFAR3 mRNA expression for predicting secondary infection in SAP patients was 0.856 (95% CI was 0.750-0.981, P < 0.001). When the optimal cut-off value was 2.37, the sensitivity was 95.83% and the specificity was 76.19%. According to the optimal cut-off value of neutrophil FFAR3 mRNA expression (2.37) for predicting secondary infection in SAP patients obtained by ROC curve analysis, 45 SAP patients were divided into two groups for subgroup analysis. It suggested that the incidence of secondary infection in SAP patients with FFAR3 mRNA expression level ≥2.37 was significantly higher than that in SAP patients with FFAR3 mRNA expression level < 2.37 [82.14% (23/28) vs. 5.88% (1/17)], and the difference was statistically significant ( P < 0.01). Conclusion:The expression of FFAR3 mRNA in neutrophils is closely related to the secondary infection in SAP patients, and monitoring its level can effectively predict the secondary infection in SAP patients.

Objective:To observe the effect of peripheral 5-hydroxytryptophan (5-HT)-induced neutrophil extracellular trap (NET) on lung injury in septic mice.Methods:Wild-type (WT type) and Tph1 knockout (KO) C57 mice (6-8 weeks) were selected and divided into WT mice sham group, WT mice sepsis group, Tph1 KO mice sham group and Tph1 KO mice sepsis group according to the random number table method. Mice in the sham group received sham surgery (only open the abdominal cavity to flip the cecum without ligation and puncture, and then close the abdominal cavity); the mice in the sepsis group received cecal ligation and puncture (CLP) to establish sepsis model. The mice were sacrificed 12 hours after the operation, and the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in bronchialalveolar lavage fluid (BALF) were detected by enzyme linked immunoadsordent assay (ELISA); at the same time, the lung tissues were collected, and the pathological changes of lung tissues were observed under light microscope, and the production of NET in lung tissues was observed by immunofluorescence microscope. Results:The pathological results suggested that the lung tissue structure in sham groups was intact without exudation, while the alveolar structures of mice in the sepsis groups were damaged, with obvious exudation in the alveolar cavity and thickened alveolar walls accompanied by a large number of inflammatory cell infiltration, and the degree of lung injury in the sepsis group of WT mice was more severe than that of the sepsis group of Tph1 KO mice. ELISA results showed that there was no statistically significant difference in the contents of TNF-α and IL-6 in mice BALF from different strains of the sham group; while the contents of TNF-α and IL-6 in BALF of septic mice group were significantly higher than those in sham group [WT mice: TNF-α (μg/L) was 158.20±28.46 vs. 14.00±3.28, IL-6 (μg/L) was 304.98±21.78 vs. 57.70±12.30; Tph1 KO mice: TNF-α (μg/L) was 85.88±20.13 vs. 14.95±1.53, IL-6 (μg/L) was 169.50±45.61 vs. 55.05±12.68, all P < 0.01], and the above index levels in the sepsis group of WT mice were significantly higher than the sepsis group of Tph1 KO mice [TNF-α (μg/L): 158.20±28.46 vs. 85.88±20.13, IL-6 (μg/L): 304.98±21.78 vs. 169.50±45.61, both P < 0.01]. Immunofluorescence staining showed that a very small amount of NET formation was detected in the mice lungs from the sham group; a large amount of NET formation was detected in the lung tissues in the sepsis group, which were significantly higher than those in sham group [WT mice: (34.75±7.27)% vs. (1.75±0.96)%, Tph1 KO mice: (14.25±5.74)% vs. (2.50±1.29)%, both P < 0.01], and the amount of NET produced in the lung tissues of the WT mice sepsis group was significantly higher than that of the Tph1 KO mice sepsis group [(34.75±7.27)% vs. (14.25±5.74)%, P < 0.01]. Conclusions:In sepsis, the increased production of inflammatory factors in the mice lung tissues induces to lung injury. The mechanism may relate to the increased production of NET in the lung tissues mediated by peripheral 5-HT synthesized by enterochromaffin cells and released into the blood; inhibiting the production of 5-HT in the peripheral blood can effectively reduce the production of NET in the lung tissues, thereby reducing lung injury.

Objective:To evaluate the value of neutrophil to lymphocyte and platelet ratio (N/LPR) for predicting 28-day mortality in sepsis patients.Methods:A retrospective analysis was conducted. The clinical data of 154 sepsis patients admitted to intensive care unit (ICU) of the Affiliated Hospital of Jiangsu University from June 2017 to June 2020 were enrolled. The time of first diagnosis of sepsis in ICU was taken as the research starting point, and the death or 28 days as the end point. The 28-day outcomes of patients were recorded. The counts of peripheral blood neutrophil (NEU), lymphocyte (LYM) and platelet (PLT) were collected from all the enrolled patients within 3 days after diagnosis of sepsis. The ratios of N/LPR and NEU/LYM (NLR) were calculated respectively. The differences of N/LPR and NLR between survival group and death group were compared. Receiver operating characteristic (ROC) curve analysis was used to analyze the value of N/LPR and NLR on predicting the 28-day mortality of sepsis patients. According to the best cut-off value of ROC curve analysis, the 28-day mortality of patients with sepsis was analyzed by subgroup analysis, and the 28-day cumulative survival of patients with sepsis was analyzed by Kaplan-Meier survival curve.Results:Of the 154 sepsis patients, the patients with age < 18 years, pregnancy, blood disease, taking aspirin or other antiplatelet drugs within 1 week, taking leucocyte drugs within 1 week, length of ICU stay < 3 days and incomplete data were excluded. Finally, 50 patients were enrolled. Among them, 30 patients survived on the 28th day and 20 died. Compared with the survival group, the levels of N/LPR and NLR in the death group were significantly increased (N/LPR: 23.85±11.99 vs. 12.41±5.25, NLR: 17.83±8.69 vs. 10.75±3.63), with statistical differences (both P < 0.01). ROC curve analysis indicated that the area under ROC curve (AUC) of N/LPR for predicting 28-day death of sepsis patients was 0.827, it was higher than that of NLR (AUC = 0.762). Base on N/LPR≥15.48 as a predictor of cut-off value of death in 28 days of sepsis patients, the sensitivity was 75.0% and the specificity was 80.0%, respectively. Base on NLR≥10.65 as a predictor of cut-off value of death in 28 days of sepsis patients, the sensitivity was 75.0% and specificity was 56.7%, respectively. Subgroup analysis showed that the 28-day mortality in the patients with N/LPR≥15.48 ( n = 21) was significantly higher than those with N/LPR < 15.48 ( n = 29; 71.4% vs. 17.2%, χ 2 = 14.901, P < 0.01); and the 28-day mortality in the patients with NLR≥10.65 ( n = 28) was also significantly higher than those with NLR < 10.65 ( n = 22; 53.6% vs. 22.7%, χ 2 = 4.884, P < 0.05). The results were consistent with Kaplan-Meier survival curve analysis. Conclusion:Peripheral blood N/LPR has a good predictive value for 28-day mortality of sepsis patients, and which is better than NLR.

Objective To evaluate combined sampling at multiple sites of gastric mucosa for Helicobacter pylori (HP) culture.Methods A total of 258 patients with upper gastrointestinal symptoms received 13C-urea breath test between August 2014 and May 2015.During endoscopy,gastric mucosa biopsy samples from the lesser curvature of the antrum (A),the greater curvature of the antrum (B),gastric angle (C) and the body of the stomach (D) were collected to isolate HP strains.The positive rates of HP based on combined sampling and single site sampling were compared with a Nemenyi test.Results Consistency between 13C-urea breath test and HP culture was 82.56％.There was significant difference between the single site sampling and two-site sampling in the positive rate of HP,except for the body of the stomach (P＜0.05).There was significant difference in the positive rate of HP between the single site sampling and three-site sampling (P ＜ 0.05).There was no significant difference between any two-site sampling in the lesser curvature of the antrum and the body of the stomach,gastric angle and the body of the stomach,the greater curvature of the antrum and the body of the stomach,and any three-site sampling (P＞0.05).Conclusion The combined sampling of the lesser curvature of the antrum and the body of the stomach have the highest cost-effectiveness in HP culture compared with the single site sampling and three-site sampling.

Objective The control rate of blood pressure in hypertension patients is very low in our country , while follow-up intervention can significantly improve the situation .This study aimed to evaluate the clinical effects of anti-hypertension under follow-up intervention . Methods From October 2013 to October 2014 , 125 patients with hypertension were chose as the study objectives after first clinical anti-hypertension and were divided into intervention group (follow-up,n=65) and control group(no follow-up,n=60). Comparative analysis was made in blood pressure control , compliance with therapy and cardiovascular event incidence between the two groups after 12 weeks'intervention. Results After 12 weeks, diastolic and systolic blood pressure in intervention group was signifi-cantly lower than that in control group (P<0.05).Significant difference was also found in the compliance with drug-taking between in-tervention group and control group (73.8%vs 43.3%, P<0.01).During the follow-up period, 1 case in the control group suffered stroke and unstable angina pectoris hospitalized for treatment . Conclusion Follow-up intervention after clinical service can improve the efficacy of blood pressure control and encourage the patients to live healthy lifestyle .

Objective To investigate the suppressive effect of exogenous carbon monoxide (CO) on abnormal platelet exocytosis and its possible molecular mechanism. Methods Venous blood was collected from healthy volunteers. Platelet-rich plasma (PRP) was isolated from the blood by differential centrifugation. The PRP was randomly divided into five groups by random number table, namely normal control group, lipopolysaccharide (LPS) group (challenged with 10 mg/L LPS), inactively exogenous carbon monoxide releasing molecule 2 (iCORM-2) group (given 10 mg/L LPS + 50 μmol/L iCORM-2 for intervention), exogenous carbon monoxide releasing molecule 2 (CORM-2) 10 μmol/L and 50 μmol/L groups (given 10 mg/L LPS + CORM-2 10 μmol/L or 50 μmol/L for intervention). After 30 minutes, enzyme linked immunosorbent assay (ELISA) was used to determine the platelet-derived growth factor BB (PDGF-BB) and matrix metalloproteinase 2 (MMP-2). Chemical fluorescein method was used to determine the platelet adenosine triphosphate (ATP). Flow cytometer was used to determine the expression of P-selectin. The expressions of Toll-like receptor 4 (TLR4), phosphorylation of protein kinase Cθ (PKCθ) and syntaxin binding protein 1 (STXBP-1) were determined by Western Bolt. The soluble N-ethylmaleimide-sensitive factor-attachment protein receptors (SNAREs) complex formation [syntaxin 2-synaptosomal-associated protein 23-vesicle associated membrane protein 8 (STX2-SNAP23-VAMP8)] mediated by STXBP-1 was determined by immunoprecipitation. Results ① Compared with normal control group, the platelet release of PDGF-BB, MMP-2 and ATP was significantly increased after LPS challenge, and the P-selectin expression of platelet was also obviously up-regulated [PDGF-BB (μg/L): 127.53±1.78 vs. 94.35±5.84, MMP-2 (ng/L): 51.87±9.20 vs. 35.83±3.17, ATP (μmol/L): 1.288±0.056 vs. 0.975±0.010, P-selectin: (3.93±0.19)% vs. (0.44±0.10)%, all P < 0.05]. The increases in platelet release of PDGF-BB, MMP-2 and ATP were suppressed by 10 μmol/L or 50 μmol/L CORM-2 administration, as well as high-expression of P-selectin in a dose-dependent manner [PDGF-BB (μg/L): 114.68±1.35, 97.08±6.14 vs. 127.53±1.78, MMP-2 (ng/L): 32.67±8.00, 24.63±1.63 vs. 51.87±9.20, ATP (μmol/L): 0.999±0.015, 0.965±0.008 vs. 1.288±0.056, P-selectin: (1.95±0.27)%, (0.94±0.11)% vs. (3.93±0.19)%, all P < 0.05]. ② Compared with normal control group, LPS challenge resulted in a significant increase in the expression of TLR4 and the phosphorylation of PKCθ and STXBP-1 [TLR4 (gray value): 1.21±0.38 vs. 0.67±0.06, p-PKCθ (gray value): 1.36±0.20 vs. 0.44±0.03, p-STXBP-1 (gray value): 1.13±0.06 vs. 0.59±0.04, all P < 0.05]. The increases in above parameters were suppressed by 10 μmol/L or 50 μmol/L CORM-2 administration in a dose-dependent manner [TLR4 (gray value): 0.76±0.05, 0.65±0.04 vs. 1.21±0.38; p-PKCθ (gray value): 0.71±0.07, 0.47±0.10 vs. 1.36±0.20; p-STXBP-1 (gray value): 0.56±0.02, 0.48±0.01 vs. 1.13±0.06, all P < 0.05]. ③ Compared with normal control group, the SNAREs proteins in platelet that combined with STXBP-1, including STX2, SNAP23 and VAMP8, were obviously increased after LPS challenge [STX2 (gray value): 1.35±0.06 vs. 0.57±0.04, SNAP23 (gray value): 0.97±0.04 vs. 0.30±0.12, VAMP8 (gray value): 1.37±0.12 vs. 0.77±0.10, all P < 0.05]. The increases in SNAREs complex formation were suppressed by 10 μmol/L or 50 μmol/L CORM-2 administration in a dose-dependent manner [STX2 (gray value): 0.77±0.02, 0.39±0.03 vs. 1.35±0.06, SNAP23 (gray value): 0.41±0.03, 0.22±0.08 vs. 0.97±0.04, VAMP8 (gray value): 0.85±0.07, 0.66±0.07 vs. 1.37±0.12, all P < 0.05]. There was no significant difference in the above mentioned parameters between iCORM-2 group and LPS group. Conclusions LPS-induced abnormal secretion of platelet was suppressed by CORM-2 administration. The mechanism may involve the TLR4/PKCθ/STXBP-1 signaling pathway activation and the SNAREs complex formation.

Objective To investigate the effect of laparoscopic cholecystectomy (LTCBDE) in treatment of patients with secondary extrahepatic bile duct stones. Methods Eighty-seven cases of our hospital patients with secondary to extrahepatic bile duct stones were randomly divided into the laparoscopic bile duct exploration and T tube drainage surgery (LCBDE) treatment group and the laparoscopic transcystic duct exploration of common bile duct lithotomy (ltcbde) treatment group. The observation focused on the operation time, bleeding volume , postoperative transfusion , postoperative drainage time , postoperative hospitalization time , cost of hospitalization, postoperative recovery time and complications compared clinical efficacies. Results LTCBDE group of patients in operation time (2.1 ± 0.5) was longer than that of the control group (1.6 ± 0.4), (P <0.001), while the bleeding volume, postoperative fluid volume, postoperative drainage time, postoperative hospitalization time, hospitalization expenses and postoperative recovery time were (17.4 ± 5.4), (6 550.4 ± 1 076.9), (3.5. 1.6), (4.1 ± 1.7), (12 243.5 ± 2 379.6), (11.3 ± 3.5) were lower than that of the group LCBDE (22.1 ± 7.5), (8 304.2 ± 1 394.8), (32.9 ± 10.4), (6.4 ± 2.4), (14 098.1 ± 2 897.3), (16.1 ± 5.7) P, respectively (P values were defined as 0.001, 0, 0, 0.015, 0.001, 0 individually); LTCBDE group of patients with bile leakage, acute peritonitis rates were 1/46,1/46,in which those were lower than the corresponding LCBDE in 6/41, 7/41 (P values were 0.033, 0.016, separately). Conclusion According to indications, LTCBD surgery has the advantages of less injury, less cost, less complications and so on. It has important significance to improve the condition of patients with secondary extrahepatic bile duct stones.

Objective To evaluate the clinical efficacy of modified Chen's biliojejunostomy technique in treating hilar cholangiocarcinoma.Methods The clinical data of the patients with hilar cholangiocarcinoma from January 2011 to June 2014 in the PLA general hospital and its first affiliated hospital were retrospective ly studied,and 17 of them underwent modified biliojejunostomy.There were 10 male and 7 female patients with a mean of 65 years old (range 34 ～82).Cases diagnosed as Bismuth-Corlette Type Ⅱ,Ⅲ a,Ⅲ b and Ⅳ were 2,6,4,and 5,respectively.Results Liver segment Ⅳ were resected from 2 patients,segment Ⅳ + Ⅴ from 7 patients and segment Ⅳ + Ⅴ + Ⅰ from 8 patients.No death was observed during the study period.One patient had mild bile leakage,2 patients had cholangititis,and another patient had biliary intestinal anastomotic bleeding.Three patients received resection and reconstruction of the portal vein.Three right hepatic arteries and 1 anterior branch of right hepatic artery were resected in combination with the tumor because of invasion.All the complications were alleviated under conservative treatment.Twelve cases had been followed up for a median time of 16 months (range 3 ～ 24).Two cases had cholangititis intermittently.One case underwent radiotherapy because of local recurrence in 2 years after the surgery.MRCP and relevant enzymes were within the normal range.Conclusion Modified Chen's biliojejunostomy is a simple,effective and safe method,which can be widely used when there are multiple biliary intestinal anastomoses.