Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.

Objective The purpose of this study was to investigate the differential expression of circRNAs in human blood, as a diagnostic marker for pre-diabetes and type 2 diabetes mellitus( T2DM). Methods Microarray analysis was used to select several differentially expressed circRNAs from three normal patients and three T2DM patients. Enlarge the sample size(normal controls,n=20;subjects with impaired glucose regulation,n=20;and type 2 diabetes mellitus,n=20) to determine a circRNA which the most evident differentially expressed by fluorescence quantitative PCR( Q-PCR). Then they were verified with expanded samples ( normal controls, n= 50; impaired glucose regulations,n=50;type 2 diabetes mellitus, n=50) by Q-PCR. Results A total of 2 953 differentially expressed circRNAs were found in microarray analysis, of which 1 439 were up-regulated and 1 514 were down-regulated. Nine differentially expressed circRNAs were selected from the 1 439 circRNAs that were up-regulated(hsa-circ-103838, hsa-circ-103965, hsa-circ-104227, hsa-circ-002117, hsa-circ-000094, hsa-circ-101226, hsa-circ-101720, hsa-circ-400029, and hsa-circ-100633). The Q-PCR results of the expanded sample( n=60) showed that the difference expression of hsa-circ-000094(Alias:has-circ-0000247) in the nine circRNAs was the most obvious one among the 3 groups, the area under the maximum curve was found by ROC curve analysis, SIGR=0. 802 5[ 95% confidence interval (0.665 5-0.939 5), P=0.001]; ST2DM=0.77[95% confidence interval (0.624-0.916), P=0.003]. In order to verify the clinical diagnostic ability of hsa-circ-000094, the experiment was conducted to further expand the sample ( n=150). The results showed that the expression of hsa-circ-000094 in the three groups was different, the difference and ROC curve analysis were statistically significant, SIGR=0. 673 3 [ 95% confidence interval (0.575 7-0. 771 0), P＜0. 01]; ST2DM=0. 723 1 [ 95% confidence interval ( 0. 632 7-0. 813 4), P＜ 0.01]. Conclusion The higher expression of hsa-circ-000094 in peripheral blood provides a certain diagnostic basis for pre-diabetes as well as type 2 diabetes mellitus.

Hypercalcemia is an abnormal calcium metabolism, which can be caused by many diseases. Hypercalcemia caused by sarcoidosis is unusual. Sarcoidosis is a systemic granulomatous disease, it is lack of specificity in initial manifestations and easy to be misdiagnosed. In this paper, we report a patient with hypercalcemia and mediastinal lymphadenectasis, which the first diagnosis was misdiagnosed as tumour. Finally, sarcoidosis was diagnosed by mediastinal lymph node biopsy, and the patient was subsequently treated with glucocorticoid. Prednisone treatment improved symptoms of sarcoidosis, normalised serum calcium and mediastinal lymph nodes were diminished. This case suggests that clinicians need to raise awareness of sarcoidosis to identify and diagnose it correctly.

OBJECTIVETo prepare streptavidin-tagged human interferon-inducible T cell alpha chemoattractant bifunctional fusion proteins (SA/hI-TAC) and evaluate its biological activity.

METHODSpET24a-SA-hI-TAC/pET21a-hI-TAC-SA plasmids were constructed and expressed in BL21. SA-hI-TAC and hI-TAC-SA fusion proteins were purified by Ni-NTA affinity chromatography, refolded by dialysis and identified by Western blotting. The bifunctionality of the fusion proteins (biotin-binding function and hI-TAC activity) was analyzed by flow cytometry and lymphocyte chemotaxis experiment, respectively.

RESULTSSA-hI-TAC/hI-TAC-SA fusion proteins were expressed at about 12% and 25% of the total bacterial protein, respectively. The two fusion proteins had a purity of about 85% and 90% after purification, and their purity reached 98% after purification with S-100 gel filtration chromatography. Both of the fusion proteins were efficiently immobilized on the surface of biotinylated mouse bladder cancer MB49 cells (91.3% for SA-hI-TAC and 98.8% for hI-TAC-SA). SA/hI-TAC induced lymphocyte chemotaxis in a dose-dependent manner, and hI-TAC-SA showed a stronger chemotactic effect than SA-hI-TAC.

CONCLUSIONSWe successfully obtained SA/hI-TAC bifunctional fusion proteins, which may potentially be used in local treatment of tumor and as a tumor vaccine.

Animals ; Biotinylation ; Blotting, Western ; Cancer Vaccines ; Cell Line, Tumor ; Chemokine CXCL11 ; chemistry ; Chromatography, Affinity ; Humans ; Interferons ; chemistry ; Mice ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; Streptavidin

Objective To prepare streptavidin-tagged human interferon-inducible T cell alpha chemoattractant bifunctional fusion proteins (SA/hI-TAC) and evaluate its biological activity. Methods pET24a-SA-hI-TAC/pET21a-hI-TAC-SA plasmids were constructed and expressed in BL21. SA-hI-TAC and hI-TAC-SA fusion proteins were purified by Ni-NTA affinity chromatography, refolded by dialysis and identified by Western blotting. The bifunctionality of the fusion proteins (biotin-binding function and hI-TAC activity) was analyzed by flow cytometry and lymphocyte chemotaxis experiment, respectively. Results SA-hI-TAC/hI-TAC-SA fusion proteins were expressed at about 12% and 25% of the total bacterial protein, respectively. The two fusion proteins had a purity of about 85%and 90%after purification, and their purity reached 98%after purification with S-100 gel filtration chromatography. Both of the fusion proteins were efficiently immobilized on the surface of biotinylated mouse bladder cancer MB49 cells (91.3% for SA-hI-TAC and 98.8% for hI-TAC-SA). SA/hI-TAC induced lymphocyte chemotaxis in a dose-dependent manner, and hI-TAC-SA showed a stronger chemotactic effect than SA-hI-TAC. Conclusion We successfully obtained SA/hI- TAC bifunctional fusion proteins, which may potentially be used in local treatment of tumor and as a tumor vaccine.

Objective To prepare streptavidin-tagged human interferon-inducible T cell alpha chemoattractant bifunctional fusion proteins (SA/hI-TAC) and evaluate its biological activity. Methods pET24a-SA-hI-TAC/pET21a-hI-TAC-SA plasmids were constructed and expressed in BL21. SA-hI-TAC and hI-TAC-SA fusion proteins were purified by Ni-NTA affinity chromatography, refolded by dialysis and identified by Western blotting. The bifunctionality of the fusion proteins (biotin-binding function and hI-TAC activity) was analyzed by flow cytometry and lymphocyte chemotaxis experiment, respectively. Results SA-hI-TAC/hI-TAC-SA fusion proteins were expressed at about 12% and 25% of the total bacterial protein, respectively. The two fusion proteins had a purity of about 85%and 90%after purification, and their purity reached 98%after purification with S-100 gel filtration chromatography. Both of the fusion proteins were efficiently immobilized on the surface of biotinylated mouse bladder cancer MB49 cells (91.3% for SA-hI-TAC and 98.8% for hI-TAC-SA). SA/hI-TAC induced lymphocyte chemotaxis in a dose-dependent manner, and hI-TAC-SA showed a stronger chemotactic effect than SA-hI-TAC. Conclusion We successfully obtained SA/hI- TAC bifunctional fusion proteins, which may potentially be used in local treatment of tumor and as a tumor vaccine.

Objective To investigate the relationship of homocysteine and blood glucose wavy coefficient with type 2 diabetic nephropathy.Methods Glycosylated hemoglobin A1c (HbA1c),fasting blood glucose(FBG),fasting C-peptide,homocysteine(Hcy),blood-fat and 24h urinary albumin quantitative (UAlb) of 154 patients with type 2 diabetes were determined,and the blood glucose wavy coefficient were calculated after blood glucose monitored by a continuous glucose monitoring system (CGMS).The patients were divided into two groups according to the quantity of UAlb:high UAlb group (n =81 ) and normal UAlb group( n =73 ).Then the difference were compared between two groups and multiple regression analysis was done between UAlb and a variety factors.Results The course of disease in high UAlb group were significantly longer than that in normal UAlb group ( (9.68 ± 7.31 ) years vs ( 5.44 ± 3.65 ) years,t =3.427,P ＜ 0.05 ).There were significant difference on HbA1c [ ( 9.61 ± 2.44 ) ％ vs ( 8.69 ± 2.35 ) ％,t =2.162 ],blood glucose wavy coefficient [ ( 3.06 ± 0.85 ) vs (2.58 ± 0.91 ),t =2.437],low density lipoprotein-cholesterol (LDL-C) [ (3.46 ± 0.83 )mmol/L vs ( 3.01 ± 0.84 ) mmol/L,t =2.596 ],UAlb [ ( 129.64 ± 118.5 ) mg/24 h vs ( 18.14 ± 3.54 )mg/24 h,t =6.421 ),UA ( ( 335.02 ± 90.39 ) mmol/L vs ( 287.00 ± 92.03 ) mmol/L,t =2.541 ) and Hcy [ ( 15.55 ± 4.53 ) mmol/L vs ( 13.12 ± 4.44 ) mmol/L,t =2.603 ] between the two groups ( P ＜ 0.05 or P ＜0.01 ).Pearson analysis showed that the courses of disease,LDL-C,UA,Hcy and blood glucose wavy coefficient were positively correlated with UAlb ( r =0.363,0.270,0.220,0.252,0.236 respectively ; P =0.000,0.008,0.033,0.014,0.022,respectively).And the multiple regression analysis indicated that UAlb was related with courses of disease(β =0.344,P =0.000),Hcy(β =0.244,P =0.011 ) and blood glucose wavy coefficient(β =0.229,P =0.012).Conclusion The elevation of serum Hcy and blood glucose wavy coefficient are risk factors to type 2 diabetic nephropathy.Lowering Hcy concentration and reducing the glucose variability may be a new way to prevent the occurrence of type 2 diabetic nephropathy.

Objective To analysis the influence factors of standardization in the hierarchical chain management of type 2 diabetes and to enhance the hierarchical chain management of type 2 diabetes.Methods ( 1 ) Six hundred and ninty patients with type 2 diabetes completed 1 years management were divided into well-controlled glycosylated hemoglobin ( HbAlc ) group (＜7.0％ ) and bad-controlled glycosylated hemoglobin (HbAlc) group ( ≥ 7.0％ ).The conditions of diet,physical activity,medication,self-blood sugar monitoring and participation in health seminars were investigated and analyzed.(2) The patients were divided into standardized management group and not standardized management group.Their age,sex,educational background,occupation,monthly income per person,medical security,the course,cognition for glycuresis,two-way transfer,and chronic complications were investigated and statistically analyzed.Results ( 1 ) The proportions of physical activity (70.1％ vs 54.2％,x2=6.163,P=0.018),self-blood sugar monitoring(60.4％ vs 43.8％,x2=6.268,P=0.016) and participation in health seminars (56.0％ vs 41.7％,x2=4.577,P=0.045) in the well-controlled HbAlc group were significantly higher than those in the bad-controlled HbAlc group.(2) Their age [(61.08 ±10.04) years old vs ( 57.75 ± 9.89 ) years old,t=2.539,P=0.012],educational background ( ratio of low educational attainment:8.3 ％ vs 17.2％,x2=6.426,P=0.041 ),medical security (own expense ratios:4.6％ vs 11.5％,x2=3.543,P=0.048 ),awareness of diabetes ( ratio of poor awareness of diabetes:19.4％ vs 41.0％,x2=17.518,P=0.000 ),two-way transfer ( ratio of not transfer treatment:4.6％ vs 14.8％,x2=7.662,P=0.022) and chronic complications ( ratio of chronic complication:41.7 ％ vs 26.2％,x2=6.130,P=0.017) were significantly different between the standardized management group and not standardized management group.(3) Logistic regression analyses indicated that the age ( OR=0.954,P=0.006),monthly income per person ( OR=4.101,P=0.018 ),medical security ( OR=7.617,P=0.003 ),cognition for glycuresis ( OR=0.030,P=0.000),two-way transfer ( OR=9.079,P=0.000) and chronic complications ( OR=0.456,P=0.031 ) were the risk factors of standardized management.Conclusion We should focus on the impact factors affecting the standardized management of patients including age,monthly income per person,medical security,awareness of diabetes,ratio of not transfer treatment,positive strategies for chronic complications,improve the hierarchical chain management of type 2 diabetes,and then make the diabetic patients to early participate in standardization management of diabetes mellitus and delay the appearance of complications.

Objective To explore the hierarchical chain management model in blood glucose control and its influence factors in patients with diabetes mellitus.Methods Health management database of diabetic patients was established in 2007 and managed by hierarchical chain management.The number of the patients reached to 1010 till 2011.The blood glucose control of diabetic patients was analyzed and its influence factors were analyzed by multivariate unconditional Logistic regression method.Results The concentration of glycosylated hemoglobin( HbA1c ) of 1010 patients with type 2 diabetes was (8.21 ±:2.70)％.Four hundred and eighty-seven cases (48.22％) reached the blood glucose standard,303 cases (30.00％)reached the blood pressure standard,245 cases (24.26％) reached the blood lipids standard,and 76 cases (7.52％) reached all three standards.Multivariate analysis showed that occupation (OR =2.521,95％ CI:1.871 - 3.397),education level (OR =1.890,95％ CI:1.642 - 2.174),disease course (OR =1.035,95％CI:1.016 -1.055),systolic pressure (OR =1.016,95％ CI:1.007 -1.025) and triglyceride (OR =1.204,95％CI:1.063 - 1.365) were the risk factors of blood glucose control (P ＜0.01).Conclusions Hierarchical chain management model is helpful for the blood glucose control in patients with type 2 diabetes.The comprehensive control and treatment of type 2 diabetes should be taken combined with related risk factors,such as blood pressure,blood lipids and diabetes disease course.

Objective To explore the hierarchical chain management model of type 2 diabetes and determine its evaluation.Method Based on the hierarchical chain management of the three community health service institutions and Dahua hospital in Shanghai Xuhui district,215 cases of type 2 diabetes had been involved in the study.Results Compared with the baseline before management,lasting blood glucose (FBG),2 h postprandial glucose (2hPBG),glycosylated hemoglobin (HbA1c),low density lipoprotein cholesterol (LDL-C),systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the diabetes after 12 months' management declined [(8.50 ±2.81) mmol/L,(11.09 ±4.01) mmol/L,(8.56 ±2.41)% ,(3.31 ± 1.06) mmol/L,(139.06 ±20.68) mm Hg (1 mm Hg = 0.133 kPa),(78.20 ± 12.11) mm Hg vs.(7.41 ±2.04) mmol/L,(9.03 ±2.46) mmol/L,(7.34 ± 1.59)% ,(3.00 ± 1.06) mmol/L,(135.48 ± 17.82) mm Hg,(77.27 ±11.83) mm Hg],and the differences were statistically significant(P＜0.01 );control rate of FBG,2hPBG,HbA1c,LDLC,SBP,DBP had improved significantly [19.5% (42/215),20.9% (45/215),24.7%(53/215),20.0%(43/215),27.4%(59/215),30.2%(65/215) vs.50.7%(109/215),53.0% (114/215),54.0%(ll6/215),42.3%(91/215),47.0%(101/215),45.6%(98/215)](P＜0.01).Conclusion Primary and secondary-care hospital based hierarchical chain management model is valid and can be implemented for type 2 diabetes.