Objective To establish a novel method for the determination of betaine in serum by high performance liquid chromatography with UV detection.Methods Pre-column derivatization of serum was performed directly in acetonitrile without extraction with p-bromophenacyl bromide and 18-crown-6 ether as catalyst.The p-bromophenacyl ester derivatives were then separated by Supelcosil LC-SCX, using an was 0.8 ml/min and the effluent was monitored at 259 nm. Betaine was used for preparation of standard curve and quantification with external standards.Results The linearity of this method was 6.25-200.00 μmol/L,the regression was 0.999 8.The detection limit was 3.0 μmol/L.The within-day imprecision was 1.88%-3.79% ( average 3.24% ), the between-day imprecision was 3.14%-6.76% ( average 4.39% ), the recovery rates were 95.89%-102.86% (average 99.16% ).Conclusion This method is sensitive, rapid, accurate and suitable for the research and routine clinical practice.

0.05).There were no signifi-cant relation between the expressions of Fas,FasL and bcl-2and IL-8.Conclusion The abnormal expressions of Fas,FasL,bcl-2may play an important role in the pathogenesis of LN.The expression of Fas and FasL may serve as active indexes for SLE and LN.The low level of IL-8may be related to the dysfunction of immunity and needs further research.

BACKGROUND: The association of tumor necrosis factor alpha (TNF-α) gene polymorphisms with the onset and development of ankylosing spondylitis (AS) has been the focus of studies on AS in the field of genetics.OBJECTIVE: To explore the association of the polymophisms of TNF-α promoter gene at positions-308 and -238 with AS susceptibility and clinical pathological changes.DESIGN: A case-control study.SETTING:The Rheumatic Immunology Department of Changhai Hospital of the Second Military Medical University of Chinese PLA.PARTICIPANTS: Totally, 108 AS patients were recruited from Rheumatic Immunology Department of Changhai Hospital, Second Military Medical University of Chinese PLA from January 1999 to December 2003 ,they had no kinship. The ratio of men to women was 5.3: 1. They aged from 13 to 71 (30-± 12) years old, and AS was divided into Ⅰ- Ⅳ radiographic stages according to the sacro-iliac joint damage. A total of 100 healthy controls were randomly selected from the blood donators(Shanghai Hospital) who were aged from 19 -56 (33 ±9) years old, and the ratio of men to women was 4.9: 1. Informed consent was obtained from all the subjects.ti-coagulated with EDTA. Polymerase chain reaction amplification and purification of the TNF-α promoter region was made and the sequence of polymerase chain reaction products was examined and displayed by Chromas 1.62 softcorresponding radiographic stage of sacro-iliac joint damage was assessed to investigate the influence of gene polymorphisms on AS.MAIN OUTCOME MEASURES: DNA direct sequencing method was used to detect -238 and -308 allele phenotypes for investigating the association with clinical presentations.G and -238G/A allele was 98.1% (106 cases) and1. 9% (2 cases) respectively in AS group and 95.0% (95 cases) and 5.0% (5 cases) respecquency of TNF-α promoter gene at positions -308. 1.1(G/G) and - 308.1.2(G/A) alleles was 82.4% (89 cases) and 17.6% (192 cases) respectively in AS group, which was not significantly different compared respectively with 85.0% (85 cases) and 14.0% (14 cases) of the control of sacro-iliac joint damage and the frequency of TNF-α promoter gene at the position of - 308 (G/G) and (G/A): AS patients with(G/G) phenotype who were confirmed of radiographic stage Ⅰ, Ⅱ, Ⅲ, and Ⅳ were observed in 3/35/40/11cases,compared with (G/A) phenotype of 1/12/6/0 cases.The difference was statistically significant (χ2GMH = 4.77, P ＜ 0.05 ).CONCLUSION: Our data suggest that the polymorphisms of TNF-α promoter gene at positions of - 238 and - 308 allele has no association with AS susceptibility, but the polymorphisms of TNF - α promoter gene at the position of -308 might exert great influence on AS according to the radiographic stage of sacro-iliac joint damage.

Objective: To explore the prevalence of attention deficit hyperactivity disorder(ADHD) and its associated factors among college students in Changsha, so as to provide evidence of for effecive intervention of ADHD. Methods: A total of 8 080 students from two universities in Changsha were investigated with self-designed general situation questionnaire, Chinese version of Wende Utah adult rating scale for ADHD, World Health Organization (WHO) Adult ADHD Scale (ASRS) and Chinese Internet Scale (CIAS-R). Results: The self-report rate of ADHD was 7.2%. Single factor analysis showed that the incidence of ADHD in gender, physical and mental illness history, family history of mental illness, relationship with parents, smoking, drinking and Internet access were statistically significont (χ2=17.68, 22.25, 43.15, 20.73, 18.61, 43.05, 22.81, 53.55, 719.65, P<0.05). Multivariate Logistic analysis showed that the younger the age, mental illness, drinking, the higher the degree of Internet dependence were positively correlated with ADHD, while gender (female) and harmony with father were negatively correlated with ADHD. Conclusion ADHD in college students can t be ignored. There are many influencing factors, suffering from mental illness, drinking and Internet addiction.

Objective:To explore the disease-related impairment of functional and structural connectivity network and their relationship with psychometric hepatic encephalopathy score (PHES) in patients with chronic hepatitis B virus-related cirrhosis (HBV-RC) by combining resting-state functional MRI (rs-fMRI) and diffusion tensor imaging (DTI).Methods:Data of 30 HBV-RC patients [including 13 HBV-RC patients with minimal hepatic encephalopathy (MHE) and 17 HBV-RC patients without MHE (NMHE)] from April 2011 to October 2011 in Guangdong No.2 People′s Hospital were analyzed prospectively, and 38 healthy individuals matched for age, sex, and education with HBV-RC patients (HC group) were included during the same period. Rs-fMR and DTI data as well as PHES data of all participants were collected. Gretna and PANDA software package were used to preprocess the imaging data and construct the functional and structural network respectively. The network-based statistic (NBS) approach was used to compare the differences of the functional and structural connections among three groups. Spearman′s correlation analysis was used to identify the relationship between functional or structural connectivity and PHES. The structural equation modeling (SEM) was used to explore the relationships among functional connectivity, structural connectivity, and PHES.Results:Compared to HC group, both functional and structural connectivity in the whole brain progressively destroyed from NMHE to MHE, mainly involving cognitive control network, default mode network, and limbic network (NBS corrected, all P<0.01). There were significantly negative relationships between functional or structural connectivity and PHES in HBV-RC patients (false discovery rate corrected, all P<0.05). The SEM results showed the influence of structural connectivity on neurocognitive impairment was mediated by functional connectivity ( P<0.05). Conclusion:Both functional and structural networks progressively destroy in HBV-RC patients as the disease advanced and these alterations significantly correlate with PHES. Besides, the influence of structural connectivity on neurocognitive impairment is mediated by functional connectivity.

Objective To explore the application value of dipstick dye immuno-assay (DDIA) for screening the schistosomiasis chemotherapy targets in the low endemic areas of Xiaogan City.Methods The residents aged 6-65 years in a village in the low endemic areas of schistosomiasis of Xiaogan City were selected and tested by the methods of fecal examination,DDIA,indirect hemagghitination (IHA),enzyme linked immunosorbent assay (ELISA) and inquiry,and the results of fecal examination were determined as the gold standard.Results The Youden' s indices of IHA,DDIA,ELISA and inquiry were 0.74,0.72,0.62 and 0.30,respectively,and the consistency rates of them were 93.38%,91.99%,81.53% and 70.03%,respectively.It took 16.70,4.95,4.12,5.63 and 2.44 Yuan screening one patient with the fecal examination,IHA,DDIA,ELISA and inquiry,respectively.Conclusion The validity of DDIA with simple operation and low cost for screening the schistosomiasis chemotherapy targets is satisfying,and the method is suitable for large scale screening in low endemic areas.

Objective:To explore the correlation between the expression of signaling lymphocyte activation molecule family 6 (SLAMF6) on peripheral blood CD8 +T cells and perforin and granzyme B and the clinical significance in patients with newly diagnosed severe aplastic anemia(SAA). Methods:The indicators of blood routine and bone marrow and peripheral blood samples of 32 newly diagnosed SAA patients admitted to Henan Provincial People′s Hospital from January 2016 to June 2019 were collected for retrospective analysis. Flow cytometry was used to detect the expression of SLAMF6, perforin and granzyme B on samples CD8 +T cell before therapy and 6 months after therapy (11 cases received transplantation, 21 cases received immunosuppressive therapy [IST]). Spearman correlation analysis was performed to determine the association between clinical indicators and laboratory test results. The expression of SLAMF6, perforin and granzyme B was also detected in 10 healthy people (normal group) and 13 myelodysplastic syndromes/paroxysmal nocturnal hemoglobinuria (MDS/PNH) patients (MDS/PNH group). Results:(1) At diagnosis: the expression of SLAMF6 was significantly lower in the SAA group than that in the normal group and the MDS/PNH group ([56.40±6.37]% vs [84.34±5.81]% and [82.24±4.98]% (both P<0.001]). The expression of perforin was significantly higher in the SAA group (32.73±8.46) than that in the normal control group (23.75%±5.10%), and the MDS/PNH group (26.12%±5.53%) (both P<0.05). The expression of granzyme B was also significantly higher in the SAA group (36.23%±7.94%) than that in the normal control group (21.67%±5.05%) and the MDS/PNH group (21.79%±5.10%) (both P<0.001). The expression of SLAMF6 was positively correlated with the hemoglobin ( r=0.804), and reticulocyte absolute values ( r=0.656) in peripheral blood, percentage of granulocytes ( r=0.643) and erythrocytes ( r=0.622) in bone marrow of SAA patients (all P<0.05). Expression of SLAMF6 was negatively correlated with perforin ( r=-0.792) and granzyme B ( r=-0.908) on CD8 +T cells in patients with SAA (both P<0.001). (2) After treatment: the expression of SLAMF6 in peripheral blood CD8 +T cells of 30 surviving patients was higher than pre-treatment ([79.19±12.69]% vs [56.40±6.37]%, P<0.001). The expressions of perforin and granzyme B were lower than pre-treatment level (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 11 transplanted patients was higher than before transplantation ([86.54±3.75]% vs [56.40±7.35]%, P<0.001). The expressions of perforin and granzyme B were lower than before transplantation (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 12 IST-respond patients was higher than that before treatment, while the perforin and granzyme B levels were lower than pre-treatment (all P<0.05). The post-treatment expressions of SLAMF6, perforin and granzyme B were similar as before treatment levels in 7 IST-unrespond patients (all P>0.05). Conclusion:SLAMF6 is significantly down-regulated on CD8 +T cells in newly diagnosed SAA, negatively correlated with the effective factors of CD8 +T cells, which might participate in the immune regulatory of CD8 +T cells as a negative regulatory factor in patients with SAA. The SLAMF6 is significantly up-regulated after hematopoietic recovery, while there is no significant change in treatment-unrespond patients, which could thus serve as an useful diagnostic and therapeutic index of patients with SAA.

Objective:To evaluate the biosafety of medical injectable carboxymethyl glycosaminoglycan gel.Methods:Ames test, chromosome aberration test in vitro and gene mutation test in vitro were used to detect the genotoxicity of the medical carboxymethyl glycosaminoglycan gel. The gel saline extract (50 ml/kg) was injected slowly through the marginal vein of the ear into Japanese big-eared rabbits. The body temperature was measured and the temperature rise was calculated. The gel saline extract (50 ml/kg) and normal saline (control) were injected intraperitoneally and intravenously into the Kunming mice, respectively. The toxicity response in mice was observed after injection, and bodyweight change was valued. The gel saline extract, normal saline and distilled water were added into the rabbit anti-clotting, to detect the rate of hemolysis.Results:Under active and inactive conditions, the number of spontaneous revertants of the 4 strains of gel saline extract group and gel DMSO extract group did not reach 2 times of that of the corresponding negative control group. The rate of chromosome aberration of the three dose groups were 0. There was no significant increase in the large colony mutation frequency, small colony mutation frequency and total mutation frequency in three dose groups (all P>0.05). After injection of gel saline extract for 24, 48 and 72 h, no toxic reaction was found in each group of mice. With the extension of time after injection, the body weight of mice in the sample group and the control group increased, but the difference was not statistically significant ( P>0.05). After injection of gel saline extract, the temperature rise of 3 Japanese big-eared rabbits were 0.0, 0.3 and 0.2 ℃ respectively. The results of hemolysis test showed that the hemolysis rate of the polycarboxymethyl glucosamine gel was 0.1%. Conclusions:No genetic toxicity changes were found in carboxymethyl glycosaminoglycan to induce gene mutation or chromosome damage in bacteria and cells, and no pyrogenicity, acute systemic toxicity and hemolysis were observed. These results indicate that thecarboxymethyl glycosaminoglycan gel has good biosafety.

Objective:To explore the clinical efficacy of reconstructing the calcaneocuboid and calcaneonavicular ligaments with the autologous semitendinosus for chronic bifurcate ligament injury.Methods:A retrospective study was conducted to analyze the clinical data of 12 patients with chronic bifurcate ligament injury who had been treated by anatomical reconstruction with the autologous semitendinosus from March 2018 to January 2021 at Department of Foot and Ankle Surgery, The Fourth Hospital of Wuhan. There were 4 males and 8 females with an age of (41.4±9.6) years. Seven left and 5 right feet were affected; the time from injury to surgery was (9.3±4.3) months. The surgical efficacy was evaluated based on anteroposterior and lateral imaging assessments of foot weight-bearing at 6 months postoperation, comparison of the visual analogue scale (VAS) pain scores for foot weight-bearing walking and the American Orthopaedic Foot and Ankle Society (AOFAS) midfoot scores between preoperation and the last follow-up, stress test for stability of the transverse tarsal joint, recovery of movement, and occurrence of complications.Results:All patients were followed up for (18.4±4.0) months. Their sports activities were completely resumed after postoperative (6.8±3.9) months. Primary wound healing was achieved in all patients. One patient suffered from cutaneous nerve lesion and skin numbness, the symptoms of which disappeared after conservative treatment. Double harmonic curves were observed on the anteroposterior and lateral imaging films of foot weight-bearing in all patients. The spaces in the calcaneocuboid and the calcaneonavicular joints were normal, the articular positions were matched, and the calcaneocuboid joint was stable without recurrence or worsening of displacement. The VAS decreased significantly from preoperative 6.5 (5.6, 7.0) to 0.0 (0.0, 1.0) at the last follow-up, and the AOFAS mid-foot score increased significantly from preoperative (55.6±8.8) points to (92.3±2.6) points at the last follow-up ( P<0.05). The stress test for the stability of the transverse tarsal joint showed joint stability. Conclusion:Reconstruction of the calcaneocuboid and calcaneonavicular ligaments using the autologous semitendinosus is an alternative treatment for patients with chronic bifurcate ligament injury, because it can restore the stability of the bifurcate ligament and obviously improve the foot and ankle functions with limited postoperative complications.

Objective: To explore the efficacies of regimens of three-drug induction therapy (ATRA+ATO+anthracyclines) versus two-drug induction therapy (ATRA+ATO) in patients with acute promyelocytic leukemia (APL). Methods: Of 184 patients diagnosed with APL from January 2009 to March 2016, 58 patients underwent three-drug induction therapy, while the rest were treated with two-drug induction therapy. Three-drug induction therapy was of ATRA (20 mg·m^-2·d^-1, d_1-28) + ATO (0.16 mg·kg^-1·d^-1, d_1-28) + Idarubicin (8 mg·m^-2·d^-1, d_3-5) /daunorubicin (40 mg·m^-2·d^-1, d_3-5) , while two-drug induction therapy ATRA+ATO with the same doses and methods as above. Of 184 cases, 69 cases accompanied with WBC counts>10×10⁹/L, 115 cases with WBC counts≤10×10⁹/L at onset. Results: ①Short-term efficacy: After one cycle induction therapy, the rates of hematologic remission, genetic remission, molecular remission and induced differentiation syndrome (DS) in three-drug regimen group were 98.3%, 87.9%, 72.4% and 0 respectively, while those in two-drug regimen group were 87.3%, 65.9%, 51.6% and 12.7% respectively. In patients with WBC >10×10⁹/L, DS rate and early mortality in three-drug regimen group were lower than in two-drug regimen group (0 vs 15.6%, 4.2% vs 15.6%, respectively). In patients with WBC≤10×10⁹/L, DS rate in three-drug regimen group was also lower than in two-drug regimen group (0 vs 12.3%) , but there were no statistical differences in terms of relapse and early mortality. ② Long-term efficacy: The relapse rate, overall survival (OS) and disease free survival (DFS) in three-drug regimen group were 0, 98.5%, 96.6% respectively, while those in two-drug regimen group were 8.6%, 86.5% and 84.1% respectively; the advantages of three-drug over two-drug regimen, especially in cases of WBC >10×10⁹/L were observed. ③ Side effects: the incidences of gastrointestinal reaction, liver dysfunction, myocardial damage and headache in three-drug regimen group hardly increased. Conclusion The efficacies of three-drug induction therapy were superior to two-drug one.