Research and exploration of salivary biological markers for hepatitis B-related hepatocellular carcinoma / 中华肝脏病杂志

Objective: To study using isotope-labeled relative and absolute quantitative proteomics methodologies to screen for salivary biological markers as a simple, non-invasive tool for identifying hepatitis B-related HCC at an early stage. Methods: Saliva samples were collected to extract salivary proteins. Isotope-labeled relative and absolute quantitative proteomics were used to analyze the differentially expressed proteins between the hepatocellular carcinoma (HCC) and non-HCC groups. Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assays were used to verify differential proteins and identify markers in liver cancer tissues and saliva. Statistical analysis was used to analyze the diagnostic efficiency of salivary biomarkers. Results: 152 differentially expressed salivary proteins were screened out between the HCC and non-HCC groups. Western blot, immunohistochemistry, and enzyme-linked immunosorbent assays validated that the expressions of α-1-acid glycoprotein 1 (ORM1) and alpha-fetoprotein (AFP) were significantly increased in HCC (P < 0.05). There was a significant correlation between salivary AFP and serum AFP (P < 0.05). HCC was diagnosed when salivary α-1-acid glycoprotein 1 combined with AFP. The area under the receiver operating characteristic curve was 0.8726 (95% confidence interval: 0.8104 ~ 0.9347), the sensitivity was 78.3%, and the specificity was 88%. Conclusion: Salivary AFP and α-1-acid glycoprotein 1 can serve as potential biomarkers for hepatitis B-related hepatocellular carcinoma.

Establishment of a reporter system for estimating activation of human hepatic stellate cells based on COL1A1 promoter and enhanced green fluorescent protein / 北京大学学报(医学版)

OBJECTIVE@#To establish a visual reporting system for evaluating the activity of collagen Ⅰ α 1 chain (COL1A1) gene promoter in immortalized human hepatic stellate cells, so as to estimate the activation status of the cells and provide a new cell model for the screening and study of anti-hepatic fibrosis drugs.@*METHODS@#The promoter sequence of human COL1A1 was amplified from the genomic DNA of human hepatocarcinoma cell line HepG2. Based on the pLVX-AcGFP1-N1 plasmid, the recombinant plasmid pLVX-COL1A1-enhanced green fluorescent protein (EGFP) was constructed, in which the enhanced green fluorescent protein gene expression was regulated by the COL1A1 promoter. The monoclonal cell line was acquired by stably transfecting pLVX-COL1A1-EGFP into the immortalized human hepatic stellate cell line LX-2 by the lentivirus packaging system and screening. The cell line was treated with transforming growth factor-β1 (TGF-β1) or co-treated with TGF-β1 and drugs with potential anti-hepatic fibrosis effects. The EGFP fluorescence intensity in cells was analyzed by the fluorescence microscope and ImageJ 1.49 software using a semi-quantitative method. The COL1A1 and EGFP mRNA were detected by reverse transcription real-time quantitative PCR (RT-qPCR), and corresponding proteins were detected by Western blot.@*RESULTS@#The recombinant plasmid pLVX-COL1A1-EGFP with the expression of EGFP regulated by COL1A1 promoter was successfully constructed. Kozak sequence was added to enhance the expression of EGFP, which was identified by double digestion and sequencing. The LX-2 monoclonal cell line LX-2-CE stably transfected with pLVX-COL1A1-EGFP was obtained. After co-treatment with TGF-β1 and 5 μmol/L dihydrotanshinone Ⅰ with potential anti-hepatic fibrosis effect for 24 h, the total fluorescence intensity and the average fluorescence intensity of LX-2-CE were lower than those in TGF-β1 single treatment group (P < 0.05), the intracellular mRNA and protein levels of COL1A1 and EGFP were also lower than those in the TGF-β1 single treatment group (P < 0.05).@*CONCLUSION@#A reporter system for estimating activation of hepatic stellate cells based on COL1A1 promoter regulated EGFP expression is successfully constructed, which could visually report the changes in COL1A1 expression, one of the activation-related markers of hepatic stellate cells, in vitro. It provides a new cell model for the screening and study of anti-hepatic fibrosis drugs.

Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models / 器官移植

Objective To evaluate the role and potential mechanism of interleukin (IL)-18/IL-18 binding protein (BP) in mediating the killing effect of natural killer (NK)-92MI cells upon endothelial cells from α-1, 3- galactosyltransferase gene-knockout (GTKO) porcine models. Methods NK-92MI cells were divided into the NK, NK+IL-18, NK+GTKO, IL-18+NK+GTKO and IL-18+IL-18BP+NK+GTKO groups. The messenger ribonucleic acid (mRNA) levels of inflammation-related genes in NK-92MI cells were detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The killing effect of NK-92MI cells on endothelial cells from GTKO porcine models was evaluated by lactate dehydrogenase (LDH) assay. The apoptosis of endothelial cells from GTKO porcine models was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. The expression levels of proteins with killing effect and apoptosis-related proteins were determined by Western blot. Results Compared with the NK, NK+IL-18 and NK+GTKO groups, the expression levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-8, IL-3, IL-6 and granulocyte-macrophage colony stimulating factor (GM-CSF) mRNA were up-regulated in NK-92MI cells in the IL-18+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Compared with the IL-18+NK+GTKO group, the expression levels of IFN-γ, TNF-α, IL-8, IL-3, IL-6 and GM-CSF mRNA were down-regulated in NK-92MI cells in the IL-18+IL-18BP+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Compared with the NK+GTKO group, the expression levels of perforin, granzyme B and IFN-γ proteins in NK-92MI cells were up-regulated, the killing rate of NK-92MI cells against endothelial cells from GTKO porcine models was enhanced, the apoptosis rate of endothelial cells from GTKO porcine models was increased, and the ratios of B cell lymphoma-2 (Bcl-2)-associated X protein (Bax)/Bcl-2 and cleaved Caspase-3/Caspase-3 in endothelial cells from GTKO porcine models were elevated in the IL-18+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Compared with the IL-18+NK+GTKO group, the expression levels of perforin, granzyme B and IFN-γ proteins were down-regulated, the killing rate of NK-92MI cells against endothelial cells from GTKO porcine models was decreased, the apoptosis rate of endothelial cells from GTKO porcine models was decreased, and the ratios of Bax/Bcl-2 and cleaved Caspase-3/Caspase-3 in endothelial cells from GTKO porcine models were declined in the IL-18+IL-18BP+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Conclusions IL-18BP may block the expression of inflammation-related genes in NK-92MI cells induced by IL-18 and the killing effect of NK-92MI cells on endothelial cells from GTKO porcine models.

Preliminary report of perioperative monitoring of six-gene-edited pig-to-cynomolgus monkey kidney xenotransplantation / 器官移植

Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K+ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β2-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.

Value of urinary α1-microglobulin and N-acetyl-β-D-glucosaminidase/urinary creatinine in monitoring early renal injury in patients with chronic hepatitis B virus-related liver diseases / 临床肝胆病杂志

Objective To investigate the value of urinary α1-microglobulin (α1-MG) and N-acetyl-β-D-glucosaminidase/urinary creatinine (NAG/UCr) in monitoring renal injury in patients with chronic hepatitis B virus (HBV)-related liver diseases. Methods A total of 85 patients with HBV-related liver diseases who attended The Second Affiliated Hospital of Kunming Medical University from August 2019 to August 2020 were enrolled, and according to the history of treatment with nucleos(t)ide analogues (NUC), they were divided into NUC treatment group with 57 patients and non-NUC treatment group with 28 patients; according to the type of NUC used, the NUC treatment group was further divided into entecavir (ETV) treatment group with 32 patients and tenofovir disoproxil fumarate (TDF) treatment group with 25 patients; according to the results of HBV serum antigen and antibody markers, the patients were divided into HBeAg-negative group with 57 patients and HBeAg-positive group with 28 patients; according to the results of serum HBV DNA quantification, the patients were divided into HBV DNA-negative group with 47 patients and HBV DNA-positive group with 38 patients; according to abdominal imaging findings, the patients were divided into non-liver cirrhosis group with 47 patients and liver cirrhosis group with 38 patients. The data on medical history and laboratory markers were collected for comparison between two groups. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of data with skewed distribution between two groups; the chi-square test was used for comparison of categorical data between two groups. The McNemar test was used to compare the diagnostic merit of each index; a Spearman correlation analysis was used to investigate the correlation of each factor with α1-MG, and NAG/UCr; the multiple linear regression analysis was used to analyze the independent influencing factors for α1-MG and NAG/UCr. Results The non-NUC treatment group, the HBeAg-positive group, and the HBV DNA-positive group had significantly higher levels of urinary α1-MG than the NUC treatment group ( Z =-2.054, P =0.04), the HBeAg-negative group ( Z =-2.293, P =0.022), and the HBV DNA-negative group ( Z =-2.229, P =0.026), respectively. The HBV DNA-positive group and the liver cirrhosis group had significantly higher levels of NAG and NAG/UCr than the HBV DNA-negative group ( Z =-2.908 and -2.824, both P < 0.05) and the non-liver cirrhosis group ( Z =-3.204 and -3.412, both P < 0.05), respectively. There was a significant difference in the proportion of patients with abnormal α1-MG and that of patients with abnormal estimated glomerular filtration rate (eGFR) (31.8% vs 20.0%, χ 2 =7.178, P =0.007), and the proportion of patients with abnormal α1-MG and NAG/UCr was significantly higher than that of patients with abnormal eGFR (35.3% vs 20.0%, χ 2 =8.049, P =0.005). There was a significant difference in diagnostic merit between α1-MG+NAG/UCr and eGFR ( P =0.015). Age ( β =0.246, P < 0.05), positive HBeAg ( β =0.284, P < 0.01), and liver cancer ( β =0.291, P < 0.01) were independent risk factors for the increase in α1-MG, while the increase in FIB-4 value ( β =0.352, P < 0.05), ascites ( β =0.260, P < 0.05), esophagogastric varices( β =-0.248, P < 0.05), positive HBV DNA ( β =0.197, P < 0.05), and high total bilirubin ( β =0.257, P < 0.05) were independent risk factors for the increase in NAG/UCr. Conclusion In patients with chronic HBV-related liver diseases, renal injury may occur during the whole course of active viral replication, liver cirrhosis, and deterioration of liver function. Antiviral therapy with NUC can alleviate renal impairment caused by HBV and is safe and reliable within a certain course of treatment. Combined measurement of urinary α1-MG and NAG/UCr has more advantages over eGFR in the diagnosis of early renal injury, and it is an effective method for renal function monitoring in patients with chronic HBV-related liver diseases.

Experimental study on α1 adrenergic receptor blocker in reducing portal hypertension in rats / 中华消化杂志

Objective:To observe whether α1 adrenergic receptor (α1AR) blocker can reduce and antagonize portal hypertension caused by α1AR activation in rats, and to provide a new approach for the clinical treatment of portal hypertension.Methods:Phenylephrine was chosen as α1AR agonist, and alfuzosin was used as α1AR blocker. The route of administration was portal vein injection, and the pressure was measured by trans-portal vein puncture. According to random number table, 32 male Sprague-Dawley rats were divided into 4 groups: control group, portal hypertension model group, alfuzosin treatment group and alfuzosin prevention group. The portal venous pressure (PVP) was measured in all rats before administration. The rats in the control group were injected with 0.9% sodium chloride solution (1 L/g), and the rats in portal hypertension model group were injected with phenylephrine(1.5 μg/g), and the PVP of the above two groups was measured again at 5 and 10 min after injection. The rats in alfuzosin treatment group were injected with phenylephrine(1.5 μg/g), PVP was measured again at 5 min after administration, and then the rats were given alfuzosin(0.9 μg/g), PVP was measured again at 5 min after administration. The rats in alfuzosin prevention group were injected with alfuzosin(0.9 μg/g), PVP was measured at 1 min after administration, and then the rats were given phenylephrine(1.5 μg/g), PVP was measured again at 1, 5 and 10 min after phenylephrine injection respectively. One way analysis of variance and Dunnett- t test were used for statistical analysis. Results:The portal vein puncture was successfully performed in 4, 6, 8 and 5 rats in the control group, portal hypertension model group, alfuzosin treatment group and alfuzosin prevention group, respectively. The PVP of rats in portal hypertension model group at 5 and 10 min after phenylephrine injection was (18.045±7.636) and (15.515±5.440) mmHg (1 mmHg = 0.133 kPa), respectively, which were both higher than that before administration ((8.452±2.830) mmHg), and the differences were statistically significant ( t=2.89 and 2.82, both P<0.05). At 5 min after alfuzosin injection, the PVP of rats in the alfuzosin treatment group was (10.088±3.743) mmHg, which was lower than that of rats at 5 min after phenylephrine injection ((16.146±4.324) mmHg) and that of portal hypertension model group at 10 min after phenylephrine injection, and the differences were statistically significant ( t=3.00 and 2.22, both P<0.05). There were no significant differences in PVP in the alfuzosin prevention group before administration, at 1 min after injection of alfuzosin, and at 1, 5 and 10 min after injection of phenylephrine (all P > 0.05). Conclusions:α1AR is an important factor involved in the regulation of PVP, and its blockers can reduce and antagonize the portal hypertension caused by α1AR activation, which is of great significance in the prevention and treatment of portal hypertension progression in liver cirrhosis.

Effects of L-Arginine and α / 中国医学科学院学报

Kidney is one of the important organs of the body.With both excretory and endocrine functions,it plays a vital role in regulating the normal physiological state.As a precursor of the nitric oxide(NO)synthesis

Significance of anti-tubulin-α-1C autoantibody in systemic sclerosis / 北京大学学报(医学版)

OBJECTIVE@#To detect the serum level of a novel autoantibody, anti-tubulin-α-1C, in patients with systemic sclerosis (SSc) and to investigate its clinical significance.@*METHODS@#Anti-tubulin-α-1C antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) in 62 patients with SSc, 38 systemic lupus erythematosus (SLE), 24 primary Sjögren's syndrome (pSS) patients, and 30 healthy controls (HCs). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), immunoglobulin A(IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), C3, C4, rheumatoid factor (RF), antinuclear antibody(ANA), anti-centromere antibodies(ACA), anticardiolipin (aCL), anti-dsDNA antibody, anti-Sm antibody, anti-RNP antibody, anti-Scl-70 antibody, anti-Ro52 antibody, anti-SSA antibody, anti-SSB antibody, centromere protein A(CENP-A), centromere protein B (CENP-B) were measured by standard laboratory techniques. Raynaud's phenomenon and modified Rodnan skin score(MRSS) were recorded to evaluate the disease status of SSc. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman rank correlation were used for statistical analyses.@*RESULTS@#The serum anti-tubulin-α-1C antibody concentration in SSc group was 81.24±34.38, the serum anti-tubulin-α-1C antibody concentration in SLE group was 87.84±38.52, the serum anti-tubulin-α-1C antibody concentration in pSS group was 59.79±25.24, and the serum anti-tubulin-α-1C antibody concentration in healthy group was 39.37±18.7. Multivariate analysis revealed that anti-tubulin-α-1C antibody levels were significantly increased in the SSc and SLE patients. The expression level of anti-tubulin-α-1C antibody in SSc was higher compared with the pSS group and the health control group (P < 0.01). Further analysis demonstrated that the elevated anti-tubulin-α-1C antibody were correlated with the SSc inflammation and disease activity markers ESR(r=0.313, P=0.019), The levels of anti-tubulin-α-1C antibody were also significantly correlated with MRSS(r=0.636, P < 0.01). The best cut-off value for the diagnose of SSc was 76.77 as mean+2SD value. The proportion of Raynaud's phenomenon was higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than that in anti-tubulin-α-1C autoantibody negative group(71.4% vs. 37.5%, P=0.039). The proportions of anti-Scl-70 antibody, anti-CENP antibody and anti-cardiolipin antibody were higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than in the anti-tubulin-α-1C autoantibody negative group (37.9% vs. 15.2%, 34.5% vs. 12.1%, 13.8 vs. 0, respectively, all P < 0.05).@*CONCLUSION@#Based on this explorative stu-dy, the level of anti-tubulin-α-1C antibody increased in the serum of the patients with SSc. There were correlations between anti-tubulin-α-1C autoantibody and clinical and laboratory indicators of the SSc patients. It may become a novel biomarker indicative of active SSc and could be applied in future clinical practice.

Effects of leptin on GABA content and receptor expression in hypothalamus of mice with sleep deprivation / 解放军医学杂志

[Abstract] Objective To observe the effects of the adipocyte hormone leptin on GABA content and receptor expression in hypothalamus of mice with sleep deprivation, and explore the possible mechanisms. Methods Male C57BL/6 mice were randomly divided into three groups (8 each): control group, sleep deprivation (SD) group and leptin supplement (L-SD) group. Mice in control group were set up in a water environment without sleep deprivation, mice in SD group were set up in a "modified multi-platform water environment" to establish a sleep deprivation model, and mice in L-SD group were given leptin 1.3 mg/kg intraperitoneally twice daily in conjunction with sleep deprivation. Seven days after sleep deprivation, the general conditions of mice were observed, body weight was measured and hypothalamic tissues and plasma specimens were collected. ELISA was used to detect the plasma leptin levels, hypothalamic γ-aminobutyric acid (GABA) and glutamate (Glu) contents. Western blotting was performed to detect the expression levels of GABA key glutamate decarboxylase 67 (GAD67) and GABAA receptor α1 subtype protein (GABAARα1). Results Compared with control group, the weight of mice in SD group significantly reduced [(22.03±0.42) g vs. (17.75±0.75) g, P0.05). The hypothalamic Glu levels were obviously higher in SD group [(686.56±10.01) ng/g] and L-SD group [(668.64+9.93) ng/g] than that in control group [(577.11±16.36) ng/g] (P0.05). The expressive levels of GAD67 and GABAARα1 protein in the hypothalamus of mice in SD group [0.68±0.06, 0.69±0.07] were significantly lower than that in control group (1.09±0.13, 0.99±0.07) (P<0.05); While the expressive levels of GAD67 and GABAARα1 proteins in the hypothalamus of mice in L-SD group (1.39±0.19 and 1.33±0.14, respectively) were significantly higher than those in SD group and control group (P<0.05). Conclusion Leptin can up-regulate the expression of the key GABA synthase GAD67, increase the content of GABA and the expression of GABAARα1 protein in hypothalamus of sleep-deprived mice, which may be an important mechanism of leptin affecting sleep.

Thymosin alpha1 protects against liver injury in rats with zymosan-induced multiple organ failure / 中国组织工程研究

BACKGROUND: Liver injury in a multiple organ failure model causes great troubles to clinicians’ medication. Thymosin α1 is used for treating chronic hepatitis and it has obvious protective effects against liver injury. OBJECTIVE: To investigate the protective mechanism of thymosin α1 on liver injury in a rat model of multiple organ failure, based on adiponectin (ADPN)/ protein kinase B (Akt)/nuclear factor κB (NF-κB) signaling pathway. METHODS: Male SPF Sprague-Dawley rats were randomly divided into four groups: normal group, model group, experimental group, and control group. Rats in the model group, experimental group, and control group were given intraperitoneal injection of 500 mg/kg zymosan (50 g/L) to construct the rat multiple organ failure model. Normal rats were injected intraperitoneally with equal doses of normal saline. Thirty minutes after the injection, the rats in the experimental group and the control group were injected intraperitoneally with 2 mL of thymosin α1 and ganlixin with the dose of 0.5 mg/kg daily, respectively. The normal group and the model group were injected intraperitoneally with the same dose of normal saline. After 7 days of continuous administration, liver function parameters were tested; histopathological changes of rat liver tissues and cell apoptosis were detected using hematoxylin-eosin staining and TUNEL staining; immunohistochemistry and western blot were used to detect the expression of interleukin-10, tumor necrosis factor α (TNF-α), adiponectin (ADPN), adiponectin recepror 2, AdipoR2, p-AKT and NF-κB. RESULTS AND CONCLUSION: Compared with the normal group, the levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin in the serum, the pathological scores of liver injury, the cell apoptotic rate, and the expression levek of TNF-α and NF-κB were significantly increased in the model group, while the serum levels of total protein, interleukin-10, ADPN, AdipoR2 and p-AKT were significantly reduced in the model group (all P < 0.05). Compared with the model group, the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, the pathological scores of liver injury, and cell apoptotic rate in the experimental group and control group were significantly reduced, and the serum levels of total protein, interleukin-10, ADPN, AdipoR2 and p-AKT were significantly increased (all P < 0.05). To conclude, thymosin α1 has a protective effect on the liver of rats with multiple organ failure induced by zymosan. The mechanism is related to the ADPN/Akt/NF-κB signaling pathway. ADPN/Akt is activated and the activation of NF-κB is inhibited, then reducing the inflammatory response.

Expression and clinical significance of α1-antitrypsin in thyroid-associated ophthalmopathy / 上海交通大学学报（医学版）

Objective : To investigate the expression and clinical significance of α1-antitrypsin (α1-AT) in tears of thyroid-associated ophthal-mopathy (TAO) patients. Methods ¡¤ Patients diagnosed with TAO at Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January to December 2019 were included and divided into active TAO group (n=33) and inactive TAO group (n=22). Thirty eyes of 30 healthy volunteers were included (n=30). Unstimulated tear samples were collected from all subjects and the concentration of α1-AT was analyzed by enzyme-linked immunosorbent assay (ELISA). Difference of α1-AT concentration among 3 groups was compared by using Kruskal-Wallis H test. Mann-Whitney U test was performed for further comparison between each 2 groups. Correlationship between α1-AT con-centration and clinical activity score (CAS) was analyzed by Spearman relation. Statistical significance was accepted at a value of P<0.05. Re-sults ¡¤ α1-AT was significantly higher in active TAO group than that in inactive TAO group and control group (both P<0.05). There was no significant difference in α1-AT between inactive TAO group and control group. Tear α1-AT level was significantly correlated with CAS (r=0.846, P=0.000). ROC curve showed that the optimal diagnostic cut off value of α1-AT was 939.48 ng/mL, and the sensitivity and specificity were 81.8% and 100% (AUC=0.959, P<0.05). Conclusion ¡¤ α1-AT is significantly elevated in the tears of active TAO patients. Tear α1-AT level has important value in the diagnosis of clinical activity of TAO.

Innovative research and clinical application of recombinant human interferon α1b in pediatrics (Volume Ⅱ) / 国际儿科学杂志

Recombinant human interferon αl b (rhIFN-1 b) is the first internationally unique novel genetic engineering medicine in China.In the previous article,it has been summarized that it is a major interferon subtype that is naturally antiviral in human body,and has high safety and broad-spectrum antiviral effect.A number of pediatric consensus and guidelines of clinical experts have formed in China's national conditions.This article summarizes its innovative research and achievements in the clinical treatment of pediatric diseases,and further discusses the problems that need to be solved in the clinical application of rhIFN-α1b in pediatrics.

Innovative research and clinical application of recombinant human interferon alpha1b in pediatrics (Vol-ume Ⅰ) / 国际儿科学杂志

Recombinant human interferonα1b (IFN-α1b) is the first internationally unique novel ge-netic engineering medicine in China. As a major interferon subtype with natural antiviral activity, IFN-α1b has high safety and broad-spectrum antiviral effect, so it has broad application prospects in the treatment of viral diseases. Chinese pediatric doctors have taken the lead globally in conducting clinical research of IFN-α1b in the treatment of respiratory syncytial virus ( RSV) pneumonia、 bronchiolitis、 hand-foot-mouth disease, her-petic angina、 viral diarrhea, etc. A series of clinical guidance and expert consensus have been published which are in line with China's national conditions. This article summarizes the innovative research of IFN-α1b and proposes common issues and potential application in pediatric clinical applications.

Management of lower ureteral stones using Tamsulosin – A prospective study

Introduction: It has been scientifically established that α1-adrenergic antagonists cause inhibition of the basal tone, peristaltic frequency, and contractions in the lower ureter. Earlier studies have shown promising results with Tamsulosin (α1-adrenergic antagonist) helping in spontaneous passage of Ureteral stones. Hence this prospective randomized double-blind placebo-controlled study is taken up to establish the real impact of Tamsulosin a specific alpha blocker (α1-adrenergic receptors) in expelling distal ureteral stones. This study aimed to include 80 patients with ureteral stones of less than 5 mm and more than 5 mm in size located in the distal ureter. Materials and methods: In the Present study, informed consent was obtained from all 80 patients and the study was conducted after institutional ethical committee approval. Patients are randomized to two groups to receive Tamsulosin and placebo along with analgesics whenever required. Patients were followed up for a period of one month to study the stone expulsion rate, drug side effects and pain episodes. Results: All 80 patients complied with prescribed treatment schedules except 4 patients in placebo group and 2 in study group who were lost to follow up. At the end of 4 weeks, stone expulsion was seen in 30 out of 38 (79% patients in study group and 20 patients out of 36 (56%) in placebo group. The stone expulsion time was shorter in the study group (6.2±3.2 days) and in 9.67±5.4 days for placebo group. No significant impact on the expulsion rate was seen in relation with age, gender and ureteric stones present either in right side or left side. The frequency of pain episodes was almost same and mild in both groups. Conclusion: Tamsulosin is safe and effective drug to enhance spontaneous passage of smaller stones present in distal ureter.

Structural characterization of polysaccharide from Polygonum multiflorum and its immunomodulatory activity / 中草药

Objective: To extract and separate a polysaccharide from Polygonum multiflorum, characterize its structural features and study its immunomodulatory activity. Methods The polysaccharide from P. multiflorum (PMT) was isolated and purified by water extraction and ethanol precipitation following Q-Sepharose Fast Flow ion exchange chromatography column. Molecular weight of PMT was determined by high performance gel permeation chromatography-multiple angle laser light scattering (HPGPC-MALLS), and monosaccharide composition was analyzed by HPLC with PMP (1-phenyl-3-methyl-5-pyrazolone) pre-column derivatization, respectively. The structure of PMT was characterized by proton nuclear magnetic resonance spectrum (2D-NMR). The immunomodulatory activities were tested by MTT, neutral red colorimetric assay and Griess method. Results: PMT was a kind of α-1,4-glucan, and its molecular mass was 3.96 × 105. PMT promoted the proliferation and phagocytosis of RAW 264.7 cells, and significantly induced the increase of NO production in a dose-dependent manner. Conclusion: The polysaccharide from P. multiflorum is a linear α-1,4-glucan with potent immunomodulatory activity, which would be potentially developed as an effective drug.

Inhalation properties of the nebulized recombinant human interferon α1b injection / 国际药学研究杂志

Objective: To investigate the aerodynamic characteristics of the nebulized recombinant human inter- feron α1b(rhIFN α1b)injection and its delivery in different respiratory modes both in vitro. Methods: The particle size distribution and aerodynamic properties of the nebulized rhIFN α1b injection for inhalation were evaluated with Spraytec STP5313 and the next generation pharmaceutical impactor(NGI). The total delivered dose and delivery rate were deter- mined using a breathing simulator. Results:After atomization, the D50 of rhIFN α1b droplets was 2.74 μm, the fine par- ticle fraction(FPF)was 77.49%, the mass median aerodynamic diameter(MMAD)was 3.26 μm, and the geometric standard deviation(GSD)was 1.93. In neonatal, infant, and child breathing modes, the delivered total amount of rhIFN α1b by spraying for 220 seconds was 2.10, 2.44, and 3.51 μg, respectively. Conclusion: After atomization, the particle size of rhIFN α1b injection was small enough to be transmitted to the lung, and the total delivered dose and delivery rate showed a tendency of increase in turn in the neonatal, infant, and child breathing modes, indicating that the effective dose of the drug and the age of patients should be considered when formulating the clinical treatment plan.

Prelimbic α1-Adrenoceptors are involved in regulation of neural activities in the amygdala in a rat model of Parkinson's disease with anxiety / 西安交通大学学报(医学版)

Objective: To observe the effects of activation or blockade of the prelimbic (PrL) α1-adrenoceptors on anxiety-like behaviors and amygdaloid neural activities in rats with Parkinson's disease (PD). Methods: The rat model of PD was established by 6-hydroxydopamine (6-OHDA) unilateral lesion of the medial forebrain bundle (MFB). Then the anxiety-like behavior of rats was detected by the open field test. In addition, the changes of anxiety-like behavior, the effects of PrL α1-adrenoceptor stimulation on monoamines and c-Fos expression in the amygdala were also observed after local injection of the selective α1-adrenoceptor agonist or antagonist into the PrL by guided cannula. Results: Unilateral 6-OHDA lesions of the MFB in rats induced anxiety-like behaviors (P<0.001). Furthermore, activation of the PrL α1-adrenoceptors significantly induced or enhanced anxiety-like behaviors in the rats (sham group: P<0.001; lesion group: P<0.05), while blockade of the α1-adrenoceptors produced anxiolytic effects (sham group: P<0.001; lesion group: P<0.05). Then activation of the PrL α1-adrenoceptors increased the levels of DA and 5-HT while blockade of the PrL α1-adrenoceptors decreased DA and 5-HT levels in the amygdala in sham-operated rats (DA & 5-HT: P<0.001). However, compared to those of sham-operated rats, activation of the PrL α1-adrenoceptors increased the levels of NA and 5-HT while blockade of the PrL α1-adrenoceptors decreased NA and 5-HT levels in the amygdala in the lesioned rats (NA & 5-HT: P<0.001). In addition, the density of c-Fos immunoreactive positive neurons in the amygdala increased after intra-PrL injection α1-adrenoceptors agonist phenylephrine (sham group & lesion group: P<0.001). Conclusion: These findings indicate that changed neural activities in the amygdala after activation or blockade of the PrL α1-adrenoceptors are involved in regulating anxiety-like behaviors in PD rats.

Effect of Tangnaikang on Imbalance Between NE and α1-AT in Spontaneous Type 2 Diabetic Mouse / 中国实验方剂学杂志

Objective: To observe the regulatory effect of Tangnaikang (TNK) on imbalance between neutrophil elastase (NE) and α1-antitrypsin (α1-AT) in ob/ob mice with type 2 diabetes mellitus (T2DM). Method: Thirty-two male SPF ob/ob mice were randomly divided into model group (DM, normal saline) and high-dose TNK group (TNKH, TNK solution 16.04 g·kg-1), middle-dose TNK group (TNKM, TNK solution 8.02 g·kg-1) and low-dose TNK group (TNKL, TNK solution 4.01 g·kg-1). Another 8 C57BL/6J mice were included in normal group (Con, saline). The experiment lasted for four weeks. The general state, body weight (BW) and fasting blood glucose (FBG) of the mice were recorded weekly, the oral glucose tolerance (OGTT) test was performed on the 25th day, the insulin tolerance (ITT) test was performed on the 27th day, and the area under the curve (AUC) was calculated. After the end of the experiment, serum was used to detect the level of fasting insulin (Fins), insulin resistance index (HOMA-IR), total triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), NE and α1-AT. Adipose tissue was used to detect the expressions of NE, α1-AT, phosphor-insulin receptor substrate 1 antibody (p-IRS1) and glucose transporter 4 (GLUT4) proteins. Result: Compared with the Con group, the BW of the ob/ob mice of the model group increased significantly, the glucose and lipid metabolism indexes showed diabetes, the serum and adipose tissue NE increased significantly (Pα1-AT decreased significantly (PPPogtt and AUCITT were significantly decreased (PPα1-AT increased significantly (PPConclusion: TNK can reduce the BW of ob/ob mice, improve glycolipid metabolism, increase α1-AT level, decrease NE level, and regulate IRS1-GLUT4 signaling pathway, which may be one of its mechanisms in improving IR of adipose tissue mediated by neutrophil.

Effects of Geraniin on Modulating the Expression of Cbfα 1 Gene in Rabbit Osteoporotic Fracture / 昆明医科大学学报

Objective To investigate the the effects of geraniin on modulating the mRNA expression of corebinding factor α 1 (Cbfα 1). Methods The m RNA expressions of Cbfα 1 were detected by real time fluorescence quantitative PCR. Results As compared with the sham group, the expressions of Cbfα 1 mRNA were markedly suppressed in the model group, 12 and 24 w after OPF (P ＜ 0.05). Compared with the model group, geraniin significantly increased mRNA expressions of Cbfα 1. Conclusion Geraniin can upregulate Cbfα 1 gene expression and improve the treatment for OPF.

Influence of high glucose on PRKAA1 expression and proliferation of trophoblast cells / 上海交通大学学报（医学版）

Objective: To explore the expression level of protein kinase AMP-activated catalytic subunit α1 (PRKAA1) in placental tissues of gestational diabetes mellitus (GDM) women, and the influence of high glucose (HG) on PRKAA1 expression and proliferation viability of trophoblast cells in vitro. Methods: The placental samples of GDM women (n=19) and normal pregnant women (n=20) of the corresponding period were collected. Real-time qPCR and Western blotting assay were used to detect the mRNA and protein levels of PRKAA1 in these biopsies, respectively. Trophoblast cells were treated by HG in vitro and then expression level of PRKAA1 was tested. CCK8 assay was used to detect proliferation viability of the cells treated by HG medium or inhibitor of PRKAA1, dorsomorphin. Results: Comparing to normal pregnant women, both mRNA and protein levels of PRKAA1 in placental tissues of GDM women significantly decreased (both P<0.05). HG treatment drastically downregulated expression of PRKAA1 in trophoblast cells in vitro (P<0.05). Both HG medium and dorsomorphin suppressed proliferation viability of trophoblast cells (both P<0.05). Conclusion: Expression level of PRKAA1 is dampened in placental tissues of GDM women. HG suppresses proliferation viability of trophoblast cells probably via downregulating PRKAA1 level in vitro.