Zinc transporter 8 (ZnT8) is an important candidate antigen for type Ⅰ diabetes. The autoantibody detection kit based on ZnT8 can be used to help diagnose type Ⅰ diabetes, and the related products have been launched in Europe and the United States. Since the recombinant production system of active ZnT8 has not been established in China, this key raw material is heavily dependent on imports. We used Saccharomyces cerevisiae to carry out the recombinant expression of ZnT8. First, multiple antigenic forms of ZnT8 were designed as C-terminal haploid (C), C-terminal diploid (C-C), and N-terminal and C-terminal concatemers (N-C). The proteins were expressed, purified and tested for antigenicity by bridging-type ELISA. The serum of 13 patients with type Ⅰ diabetes and the serum of 16 healthy volunteers were detected. C, N-C, and C-C proteins had similar detection rates, which were 53.8% (7/13), 61.5% (8/13) and 53.8% (7/13). The specificity of the three groups was 100% (16/16). The detection value on positive samples P3, P4, and P8 increased by more than 90%, indicating better serum antibody recognition ability. Finally, N-C protein was selected for further serum sample testing, and the test results were characterized by receiver operating characteristic (ROC) curve for sensitivity and specificity. Compared with imported gold standard antigen, the sensitivity was 76.9% (10/13) and the specificity was 87.5% (14/16). There was no significant difference in the sensitivity of the method, but the specificity needed to be improved. In conclusion, the ZnT8 N-terminal and C-terminal concatemer protein developed based on S. cerevisiae expression system is expected to be a key alternative raw material in the development of in vitro diagnostic reagents for type Ⅰ diabetes.
Antigens ; Autoantibodies ; Diabetes Mellitus, Type 1/diagnosis* ; Enzyme-Linked Immunosorbent Assay ; Humans ; Saccharomyces cerevisiae/genetics* ; Zinc Transporter 8/genetics*

【Objective】 To evaluate the suitability of current quality standard for end-stage hemolytic rate required by Quality Requirements for Whole Blood and Blood Components in Taiyuan through a 5- year retrospective analysis of leukocyte-reduced RBCs in additive solution in Taiyuan Blood Center. 【Methods】 A total of 240 packs of leukocyte-reduced RBCs in additive solution were collected from January 2007 to December 2021. Hb concentration, hematocrit(Hct) and free Hb in supernatant at end-stage were detected to measure the hemolytic rate. And the hemolytic rate was then compared with the current standard(0.8%) to analyze whether statistical differences existed. 【Results】 The hemolytic rate of leukocyte-reduced RBCs in additive solution were 0.109±0.123 in 2017, 0.113±0.085 in 2018, 0.121±0.076 in 2019, 0.101±0.057 in 2020, and 0.128±0.089 in 2021, with no significant difference among each other (P>0.05). And 97.5% of the products showed an end-stage hemolytic rate below 0.4%, 0.83% among 0.6%~0.7%, and none over the standard of 0.8%. 【Conclusion】 The current national standard for end-stage hemolytic rate of leukocyte-reduced RBCs in additive solution might not be suitable for blood quality control in Taiyuan, and a much stricter criteria should be considered.

Objective:To investigate the role of LncRNA-TUG1 in the injury of intestinal epithelial cells induced by lipopolysaccharide (LPS).Methods:LPS was used to treat HIEC-6 human intestinal epithelial cells for 24 h to construct a sepsis injury model. Whole transcriptome RNA sequencing was used to analyze the expression changes of mRNA, microRNA and lncRNA in HIEC-6 cells after LPS treatment. Real-time fluorescence quantitative (qRT-PCR) and Western blot was performed to detect the expression changes of lncRNA-TUG1, microRNA-132-3p (miR-132-3p), SIRT1 mRNA and SIRT1 protein in HIEC-6 cells after LPS treatment. The expression levels of LncRNA-TUG1, miR-132-3p and SIRT1 were artificially changed by in vitro transfection. qRT-PCR and Western blot were used to confirm the regulatory effect of lncRNA-TUG1 on microRNA-132-3p and SIRT1. CCK-8 and flow cytometry were used to analyze the effects of LncRNA-TUG1, miR-132-3p and SIRT1 on the proliferation and apoptosis of HIEC-6 cells. The dual luciferase report analysis was used to verify the targeting relationship between LncRNA-TUG1, miR-132-3p and SIRT1. Statistical analysis was performed using SPSS 17.0, and differences between the two groups were compared using independent sample t test. Results:RNA sequencing results showed that the expressions of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells after LPS treatment ( t=3.26, P<0.05 and t=2.55, P<0.05), but the expression of miR-132-3p was increased ( t=4.12, P<0.05). In vitro cell experiments, the expression of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells treated with LPS ( t=5.69, P<0.05 and t=5.712, P<0.05), while the expression of miR-132-3p was increased ( t=3.88, P<0.05). Overexpression of lncRNA-TUG1 increased the proliferation rate ( t=6.55, P<0.05) and decreased the apoptosis rate ( t=3.94, P<0.05) of LPS-treated cells. Upregulation of lncRNA-TUG1 decreased the expression of miR-132-3p ( t=4.66, P<0.05), and increased the mRNA and protein levels of SIRT1 ( t=3.91, P<0.05). Transfection of miR-132-3P mimic could inhibit the mRNA ( t=4.08, P<0.05) and protein levels of SIRT1. In LPS-treated cells, the cells co-transfected with miR-132-3pmimic and siRNA-SIRT1 had a lower proliferation rate ( t=4.55, P<0.05 and t=5.67, P<0.05) and a higher apoptosis rate ( t=3.90, P<0.05 and t=4.22, P<0.05) than those transfected with only pcDNA3.1-lncRNA-TUG. Conclusions:lncRNA-TUG1 may act as a ceRNA to regulate miR-132-3p/SIRT1, therefore alleviating HIEC-6 cell injury caused by LPS. Intervention of lncRNA-TUG1/miR-132-3p/SIRT1 regulatory pathway may become a potential strategy to prevent sepsis-induced intestinal mucosal damage.

Objective To evaluate the efficacy of 0.3％ sodium hyaluronate ophthalmic solution in mild-to-moderate dry eye patients.Metbods A prospective,multicenter,and self-controlled clinical trial was performed on 200 patients who were diagnosed as mild-to-moderate dry eye from January 2015 to June 2017 in Eye Institute of Xiamen University,Eye & ENT Hospital of Fudan University,Zhongshan Ophthalmic Center,Xiangya Hospital Central South University,Eye Hospital of Wenzhou Medical University,and Beijing Tongren Hospital,Capital Medical University.The study protocol was approved by the Ethics Committees of Eye Institute of Xiamen University,written informed consent was obtained from each patient prior to any medical examination.All patients were treated with 0.3％ sodium hyaluronate ophthalmic solution 6 times per day (one drop each time) for 28 days.Corneal fluorescein sodium staining,tear film break-up time (BUT),Schirmer Ⅰ test (S Ⅰ t),degree of conjunctival hyperemia,eyelid margin,meibomian gland secretion,secretory capacity of meibomian gland and subjective symptoms were assessed at baseline,on the 14th day and 28th day after treatment.Bulbar impression cytology was evaluated at baseline and on the 28th day after treatment.Irritation of 0.3％ sodium hyaluronate ophthalmic solution was estimated on the 14th day and 28th day after treatment.Results The total score of subjective symptoms,BUT,S Ⅰ t,degree of conjunctival hyperemia were significantly different among different treatment time points (F =108.969,27.598,16.838,36.750;all at P＜0.01).Compared with before treatment,the total score of subjective symptoms was significantly decreased,the degree of conjunctival hyperemia and the total corneal fluorescein sodium staining point number were significant decreased on the 14th day and 28th day after treatment.The total score of subjective symptoms,degree of conjunctival hyperemia and total corneal fluorescein sodium staining point number on the 28th day after treatment were significant lower than those on the 14th day after treatment.Compared with before treatment,the BUT was significantly longer and the S Ⅰ t scores were significantly increased on the 14th day and 28th day after treatment.The BUT on the 28th day after treatment was significantly longer than that on the 14th day after treatment;no significant difference in S Ⅰ t was observed between the 28th day and the 14th day after treatment.The scores of palpebral margin change,meibomian gland secretory ability and secretion characteristics were not significantly different among different treatment time points(H=0.255,2.356,0.294;all at P＞0.05).The impression cytology grade on the 28th day after treatment was 1.08±0.74,which was significantly lower than 1.53±0.76 before treatment (t =5.979,P＜0.01).The number of goblet cells on the 28th day after treatment was significantly higher than that before treatment (U =1 806.500,P＜ 0.01).On the 14th day after treatment,70％ of the patients indicated that the drug was non-irritating,and no patient had intolerable irritation affecting daily lives.All patients had good tolerance to this drug.Conclusions The use of 0.3％ sodium hyaluronate eye drops can improve the symptoms and signs of mild to moderate dry eye,which can be widely used for mild-to-moderate dry eye patients in clinic.

Objective To explore cognitive impairment and related factors in patients with Wilson disease (WD) and to screen the risk factors of cognitive impairment in order to provide evidence for clinical intervention. Methods The Chinese Version Addenbrooke＇s Cognitive Examination-III (ACE-III-C) was used to assess the cognitive function. The WD patients with cognitive impairment were analyzed the difference between those with non-cognitive disorders in the Young scale, Baethel scale and biochemical indicators. Risk factors for cognitive impairment in WD patients were analyzed by multiple linear regression. Results Cognitive impairment occurred in 43 (59.7%) of 72 patients with WD. ACE-III-C total score, attention, memory, language fluency, visual spatial factor scores, Young scores, Barthel scores and serum copper levels were significantly different between patients with cognitive impairment and patients with non-cognitive impairment (P＜0.01). Linear regression analysis showed that serum copper levels were the most important risk factors for ACE-III-C total score and cognitive subfields (P＜0.01). Serum zinc levels as a secondary risk factor of language fluency and visual space (P＜0.05). Age-related participation affected language fluency (P＜0.05). Conclusions Serum copper and zinc levels may be the main risk factors of cognitive impairment. Modulation of serum copper and zinc levels may be the key for intervention to treat cognitive impairment in WD patients.

Objective To get an understanding of the patient experience in public hospitals nationwide, and to evaluate the implementation of the Action Plan to Improve Health Care. Methods Supported by the mobile technology, from September 6, 2017 to December 15, 2018, the authors conducted an online survey that measured the satisfaction of both inpatients and outpatients at secondary and tertiary hospitals across the country. 15 questions from six dimensions including registration experience, patient-doctor communication, nurse-patient communication, the healthcare signage system, responsiveness of care providers and privacy protection were prepared for outpatients, while 20 questions from nine aspects such as nurse-patient communication, patient-doctor communication, pain management, medication communication, admission and discharge information, responsiveness of care-givers, food service, friendliness to patient family, and the healthcare signage system were directed at inpatients. Descriptive statistical analysis was used to describe the basic features of the data. Results 9.18 million valid responses from outpatients and another 5.38 million from inpatients were obtained. The overall satisfaction rate with outpatient services had reached a score of 90.45 points where nurse-patient communication stands out as the top-rated dimension and privacy protection gets the lowest rating. On the other hand, the inpatient satisfaction stands at a score of 93.01 with friendliness to patient family receiving the top score and patient-doctor communication the lowest. Conclusions Despite the positive feedback Chinese patients give on the outpatient care they receive, we should make efforts to improve the outpatient care environment, the wayfinding system, privacy protection, and responsiveness of care-givers.

Objective: To understand the self-health management ability of college students of Zhuang nationality in the economically underdeveloped areas of Guangxi and assoliated factors, and to provide support for improving the health of college students in economically underdeveloped areas. Methods: Five colleges and universities were selected in the economically underdeveloped areas of Guangxi, and 1 042 college students of Zhuang nationality were randomly selected to conduct self-health management ability surveys, and the influencing factors affecting self-health management ability were analyzed. Results: The self-health management ability of Zhuang college students in the economically underdeveloped areas of Guangxi scored (3.63±0.59) points. The scores of boys’ self-health management behaviors were higher than those of girls, which was statistically significant (t=2.92, P<0.01). There was a statistically significant difference in the total scores of cognitive and self-health management among students of different grades (F=8.03, 3.04, P<0.01). The dimensions and total scores of self-health management of medical students were higher than those of other majors, and there was statistical significance (P<0.01). The scores of behavior, environment and total scores of students with different self-assessment status were statistically significant (P<0.01). Regression analysis showed that professional, gender, and self-rated health were the main influencing factors of behavior and environment (P<0.01); gender, grade, and major were the main influencing factors of cognition (P<0.01); grade, professional, self-evaluation health were the main influencing factors of the total score of self-health management (P<0.01). Conclusion The Zhuang nationality students in the economically underdeveloped areas of Guangxi have higher self-health management ability, but their behaviors are not very good. They should be intervened and guided by different groups to improve the self-health management ability of Zhuang college students.

Objective To evaluate the efficacy of 0. 3％ sodium hyaluronate ophthalmic solution in mild.to. moderate dry eye patients. Methods A prospective,multicenter,and self.controlled clinical trial was performed on 200 patients who were diagnosed as mild.to.moderate dry eye from January 2015 to June 2017 in Eye Institute of Xiamen University,Eye &ENT Hospital of Fudan University,Zhongshan Ophthalmic Center,Xiangya Hospital Central South University, Eye Hospital of Wenzhou Medical University, and Beijing Tongren Hospital, Capital Medical University. The study protocol was approved by the Ethics Committees of Eye Institute of Xiamen University,written informed consent was obtained from each patient prior to any medical examination. All patients were treated with 0. 3％ sodium hyaluronate ophthalmic solution 6 times per day ( one drop each time) for 28 days. Corneal fluorescein sodium staining,tear film break.up time (BUT),SchirmerⅠtest (SⅠt),degree of conjunctival hyperemia,eyelid margin,meibomian gland secretion,secretory capacity of meibomian gland and subjective symptoms were assessed at baseline,on the 14th day and 28th day after treatment. Bulbar impression cytology was evaluated at baseline and on the 28th day after treatment. Irritation of 0. 3％ sodium hyaluronate ophthalmic solution was estimated on the 14th day and 28th day after treatment. Results The total score of subjective symptoms,BUT,SⅠt,degree of conjunctival hyperemia were significantly different among different treatment time points( F=108. 969,27. 598,16. 838,36. 750;all at P<0. 01). Compared with before treatment,the total score of subjective symptoms was significantly decreased, the degree of conjunctival hyperemia and the total corneal fluorescein sodium staining point number were significant decreased on the 14th day and 28th day after treatment. The total score of subjective symptoms,degree of conjunctival hyperemia and total corneal fluorescein sodium staining point number on the 28th day after treatment were significant lower than those on the 14th day after treatment. Compared with before treatment,the BUT was significantly longer and the SⅠt scores were significantly increased on the 14th day and 28th day after treatment. The BUT on the 28th day after treatment was significantly longer than that on the 14th day after treatment;no significant difference in SⅠt was observed between the 28th day and the 14th day after treatment. The scores of palpebral margin change,meibomian gland secretory ability and secretion characteristics were not significantly different among different treatment time points(H=0. 255,2. 356,0. 294;all at P>0. 05). The impression cytology grade on the 28th day after treatment was 1. 08±0. 74,which was significantly lower than 1. 53±0. 76 before treatment (t=5. 979, P<0. 01). The number of goblet cells on the 28th day after treatment was significantly higher than that before treatment(U=1806. 500,P<0. 01). On the 14th day after treatment,70％ of the patients indicated that the drug was non.irritating,and no patient had intolerable irritation affecting daily lives. All patients had good tolerance to this drug. Conclusions The use of 0. 3％ sodium hyaluronate eye drops can improve the symptoms and signs of mild to moderate dry eye,which can be widely used for mild.to.moderate dry eye patients in clinic.

Although oxymatrine (OMT) has been shown to directly inhibit the replication of hepatitis B virus (HBV), limited research has been done with this drug. In the present study, the antiviral effect of OMT was investigated in an immunocompetent mouse model of chronic HBV infection. The infection was achieved by tail vein injection of a large volume of DNA solution. OMT (2.2, 6.7 and 20 mg/kg) was administered by daily intraperitoneal injection for 6 weeks. The efficacy of OMT was evaluated by the levels of HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B core antigen (HBcAg). The immunoregulatory activity of OMT was evaluated by serum ELISA and flow cytometry. Results shows that OMT at 20 mg/kg inhibited HBV replication, and it was more efficient than entecavir (ETV) in the elimination of serum HBsAg and intrahepatic HBcAg. In addition, OMT accelerated the production of interferon-(IFN-) in a dose-dependent manner in CD4T cells. Our findings demonstrate the beneficial effects of OMT on the enhancement of immunological function and in the control of HBV antigens. The findings suggest this drug to be a good antiviral therapeutic candidate for the treatment of HBV infection.

Objective:To establish mice models of Staphylococcus aureus bloodstream infections so as to investigate the inflammatory responses and histopathological changes in bloodstream infections (BSIs) mice.Methods:C57BL/6 mice were inoculated with S.aureus intravenously or intraperitoneally to induce BSIs.Survival rate , weight loss and murine sepsis scores ( MSS ) were ob-served.Blood samples and tissue homogenates were plated on agar to determine bacterial burden .Inflammatory proteins ( CRP,PCT) and cytokines ( IL-1β, IL-6 and TNF-α) were determined by ELISA kits.Histopathologic changes were also assessed by pathological inflammation scores(PIS),macroscopic and microscopic examination.Results: About 70% survival rate was observed in 4.5×108 CFU/ml S.aureus induced BSIs mice.Body weight decreased and sepsis scores increased significantly since 24 h post-infection in BSIs mice,and more prominent in IV group.The counts of WBC began to significantly increase at 3 h post-infection,while CRP and PCT levels peaked at 48 hours in IV and IP groups ( 60.80 ±5.63 vs 40.58 ±7.54 for CRP;6.796 ±1.16 vs 2.740 ±0.36 for PCT ) . Moreover,the levels of IL-1β,IL-6 and TNF-αin serum and tissue homogenates ( liver,lungs,and kidneys ) were significantly elevated in BSIs mice.Pathological changes in tissues (liver,lungs and kidneys) and higher pathological inflammation scores (PIS) were also observed in BSIs mice.Conclusion:Our study represents an effective approach for S.aureus BSIs model to mimic human sepsis.Our results demonstrated that inflammation protein (PCT,CRP) and cytokines(IL-6,IL-1βand TNF-α) play an important role in the in-flammatory response and histopathological changes during BSIs caused by S .aureus.