Objective A enzymatic method for measurement of calcium in serum by using urea amidolyase was introduction. Methods Double reagent two-point method.TEA buffer(pH 8.0,200 mmol/L)was used as the buffer solution.The first reagent consisted of sodium bicarbonate 26.0 mmol/L,NADPH 0.4 mmol/L,2-oxoglutaric acid 8.0 mmol/L,urea amidolyase 115 U/L and GLDH 43.0 kU/L.The second reagent contained sodium bicarbonate 26.0 mmol/L and ATP 6.2 mmol/L in the buffer solution.Results The Linearity range was 0.5～5.0 mmol/L.The average rate of recovery was 102.7%.The average within-run and between-run CV were 2.13% and 2.69%.Camparison with OCPC(X1)，showed that Y1=1.02X1-0.021,r=0.981,and with Arsenazo Ⅲ(X2),Y2=0.99X2+0.03,r=0.978.No interference from as much as 300 μmol/L of bilirubin,4 g/L of hemoglobin,2.5 mmol/L of magnesium,2.0 mmol/L of ammonium ion and 8.0 mmol/L of triglyceride.Conclusion This method was simple,accurate and ripid,suitable for automatic analysis of calcium in serum.

Objective : To analyze the characteristics of HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou, and the influencing factors for new infection and local infection, so as to provide the evidence for AIDS prevention and control. Methods : From 2017 to 2019, the newly reported HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou were recruited, and their demographic information, previous sexual behaviors and history of HIV testing were collected in the questionnaire survey. The multivariate logistic regression model was used to analyze the influencing factors for new infections and local infections. Results : A total of 522 participants from 668 newly reported HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou during this period were surveyed．Among 522 cases, 263 ( 50.38% ) were aged 40 years or above, 218 ( 41.76% ) were married, 326 ( 62.45% ) had an educational level of junior high school or below, and 340 ( 65.13% ) were not local. Among 504 cases whose infection time could be determined, 72 ( 14.29% ) were newly infected within one year; age of 40 years below ( OR=4.148, 95%CI: 1.956-8.795 ), history of HIV testing ( OR=2.049, 95%CI: 1.163-3.609 ) and history of sexually transmitted diseases ( OR=2.169, 95%CI: 1.076-4.374 ) were risk factors for new infection. Among 454 cases whose infection location could be determined, 267 ( 58.81% ) were infected in Hangzhou; educational level of high school or below ( OR=2.538, 95%CI: 1.252-5.145 ) , Hangzhou residence ( OR=7.835, 95%CI: 4.227-14.353 ), living in Hangzhou for a year or over ( OR=18.960, 95%CI: 8.755-41.060 ) and monthly income of 3 000 yuan or over ( OR=2.630, 95%CI: 1.546-4.474 ) were risk factors for local infection. Conclusions The HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou are mainly floating population and less educated. The newly infected cases are more likely to be young and middle-aged people and patients with sexually transmitted diseases, the locally infected cases are more likely to be people with permanent residence, less educated and high income.

Objective : To investigate the quality of life among people living with HIV/AIDS in Hangzhou City and analyze the influencing factors, so as to provide insights into the control of AIDS. Methods : From 1 January 2014 to 30 June 2018, the demographic characteristics, medical expenditures and disease status were collected from HIV/AIDS patients living in Hangzhou City, and the quality of life was assessed using the simplified Chinese version of Medical Outcomes Study-HIV Health Survey ( MOS-HIV ). Factors affecting the quality of life were identified among HIV/AIDS patients using multivariable linear regression analysis. Results : A total of 2 808 HIV/AIDS patients were surveyed, including 1 684 cases with HIV infections and 1 124 cases with AIDS. The participants included 2 510 men ( 89.39% ) and 298 women ( 10.61% ), and were predominantly at ages of 25 to 39 years ( 1 531 cases, 54.52% ). The physical and mental health scores were 53.87±6.96 and 46.03±9.09, respectively. Multivariable linear regression analysis identified age, average monthly income, self-paid medical expenses during the past year, and the latest CD4+T cell count as factors affecting physical and mental health ( P<0.05 ). Conclusions The quality of life is low among people living with HIV/AIDS in Hangzhou City, and is associated with age, income, medical expenditures and CD4+T cell count.

Objective: To investigate the immune reconstitution of HIV/AIDS patients and its influencing factors after receiving antiviral therapy (ART) in Hangzhou City, so as to provide insights into improving the treatment effects and quality of life in HIV/AIDS patients. Methods: A retrospective cohort of HIV/AIDS patients who began antiviral treatment between January 1, 2016 and August 31, 2021 and had a baseline CD4+T lymphocyte (CD4) counts of less than 500 cells/μL or a baseline CD4/CD8+T lymphocyte (CD8) ratio of less than 0.8 in Hangzhou City was followed up until August 31, 2023. Demographic information, antiviral therapy in formation, CD4 counts, and CD4/CD8 were collected from the Chinese Disease Prevention and Control Information System. A good immune reconstitution was defined as having CD4≥500 cells/μL and CD4/CD8≥0.8. The immune reconstitution status of HIV/AIDS patients were analyzed, and factors affecting immune reconstitution were identified using a multivariable Cox proportional risk regression model. Results: A total of 3 349 HIV/AIDS patients were enrolled, with a median age at ART of 31 (interquartile range, 20) years. There were 3 075 males (91.82%), 1 600 cases with college education and above (47.78%) and 2 455 cases at WHO clinical stage Ⅰ-Ⅱ(73.31%). There were 1 368 cases with good immune reconstitution, accounting for 40.85%, and the proportion of HIV/AIDS patients with good immune reconstitution that began ART in 2016 was the highest, reaching 51.90%. Multivariable Cox proportional risk regression model identified WHO clinical stage (Ⅰ-Ⅱ, HR=2.529, 95%CI: 2.023-3.162), timely ART (HR=1.196, 95%CI: 1.027-1.394), initial treatment regimen (TDF+3TC+NVP/EFV, HR=2.185, 95%CI: 1.891-2.524; integrase inhibitors, HR=8.509, 95%CI: 6.706-10.795), baseline CD4/CD8 (≥0.1, HR: 1.600-4.515, 95%CI: 1.061-6.661), baseline hemoglobin (<90 mg/dL, HR=0.327, 95%CI: 0.121-0.880), hepatitis B infection (HR=0.619, 95%CI: 0.457-0.840) and hepatitis C infection (HR=0.308, 95%CI: 0.099-0.956) as factors affecting immune reconstitution in HIV/AIDS patients. Conclusion The immune reconstitution in HIV/AIDS patients after ART is associated with WHO clinical stage, timely ART, initial treatment regimen, baseline CD4/CD8, baseline hemoglobin and hepatitis B or C infection.

A new enhancement method is proposed based on the characteristics of fundus images in this paper. Firstly, top-hat transform is utilized to weaken the background. Secondly, contrast limited adaptive histogram equalization (CLAHE) is performed to improve the uneven illumination. Finally, two-dimensional matched filters are designed to further enhance the contrast between blood vessels and background. The algorithm was tested in DIARETDB0 databases and showed good applicability for both normal and pathological fundus images. A new no-reference image quality assessment method was used to evaluate the enhancement methods objectively. The results demonstrated that the proposed method could effectively weaken the background, increase contrast, enhance details in the fundus images and improve the image quality greatly.
Algorithms ; Contrast Media ; Fundus Oculi ; Humans ; Image Enhancement

Objective To evaluate the effects of PM2.5 on the differentiation of splenic CD4 + T lymphocytes in acute asthma mice.Methods (1) Mouse models of acute asthma were established by ovalbumin (OVA) sensitization and challenge.(2) PM2.5 was collected in the urban area of Zhanjiang city under heavy traffic and serious air pollution from total suspended particulate(TSP) mid-flow sampler and multistage particles cutters and the dry powder of PM2.5 was prepared.(3) Specific-pathogen free Balb/c mice,female,at 6 to 8 weeks of age were randomly divided into 8 groups (8 mice each group):a negative control group (NC group),asthma control group (AC group),sensitized mice treated with different doses of PM2.5 groups (SP groups) and asthmatic mice treated with different doses of PM2.5 groups (AP groups).SP groups and AP groups were respectively divided into 3 subgroups according to the dose of PM2.5.The AC group,SP groups and AP groups were sensitized on D0,D7 and D14,and the NC group was treated with NS as controls.The SP1/AP1 group,SP2/AP2 group and SP3/AP3 group were respectively given 50 μL PM2.5 suspension.NC group and AC group were instilled with NS as controls.AC group and AP groups were challenged by aerosol of OVA,and NC group and SP group were treated with NS as controls.Twenty-four hours after last challenge,all the mice were sacrificed,and the percentage of regulatory T cells (Treg),T helper cell type 1 (Th1),Th2 and Th17 of splenic CD4 + T lymphocytes was detected by flow cytometry (FCM).Results (1) An OVA-induced mouse models with acute asthma were successfully established.(2) Comparison of the percentage of Treg of splenic CD4 + T lymphocytes:SP group [(12.28 ± 0.73) ％,(11.93 ± 0.81) ％ and (11.70-± 1.14) ％] and AC group [(12.18 ± 1.00) ％] were lower than that in the NC group[(13.50 ± 0.39) ％] (P ＜ 0.05),AP3 group [(10.58 ± 0.65) ％] was lower than that in the AC group and AP1 group [(11.91 ± 0.79) ％] (P ＜ 0.05).(3) Comparison of the ratio of Th1/Th2 of splenic CD4+ T lymphocyte:SP1 group [(7.74 ± 1.21)％] was higher than that in the NC group [(5.52 ± 1.06) ％] (P ＜0.05),SP2 group[(6.30 ±0.58) ％] was lower than that in the SP1 group(P ＜0.05),SP3 group [(4.87 ± 0.82) ％] was lower than that in the SP2 group (P ＜ 0.05);AC group [(3.69-± 0.47) ％] was lower than that in the NC group and SP3 group (P ＜ 0.05);AP3 group [(2.92 ± 0.57) ％] was lower than that in the AC group(P ＜ 0.05).(4) Comparison of the percentage of Th17 of splenic CD4+ T lymphocyte:AP3 group [(1.46 ± 0.39) ％] was higher than that in the NC group [(0.89 ± 0.24) ％] and the AP2 group [(0.83 ± 0.15) ％] (P ＜ 0.001).Conclusions PM2.5 can inhibit splenic CD4 + T lymphocyte of acute asthma mice differentiation into Treg and Th1,and promote their differentiation into Th2 and Th17,through which aggravates inflammation reactions in the airway.

Objective To investigate the change of serum remnant like particle cholesterol (RLP C) and the association of RLP C with diabetic nephropathy (DN) in the patients with type 2 diabetes mellitus (DM). Methods Serum RLP C concentration was assayed by immunoseparation method. Plasma ? granule membrane glycoprotein 140 (GMP140) and nitric oxide (NO) levels were also measured. Urinary albumin excretion rate (UAER) was measured by radioimmunoassay, and the patients with type 2 DM were assigned to three groups according to UAER 〔35 without DN (D 1 group, UAER200 ?g/min)〕. Results Serum RLP C concentration in 96 patients with type 2 DM was significantly increased compared to that in 44 normal controls (NC) 〔(0.281?0.162)mmol/L vs (0.193?0.125)mmol/L, P

Objective To explore the relationship of protein kinase B1 ( PKB1 ) gene polymorphisms in PI3-K pathway of BDNF and event-related potentials in depression.Methods The design of case-control research was used ,and 91 major depressive patients and 65 normal controls who were made in age and gender matched with patients were measured auditory event-related potential P300 and contingent negative variation ( CNV ) in the day when two groups were collected.Polymerase chain reaction (PCR) and direct DNA sequencing technology were used to detect PKB1 gene polymorphisms.Three SNPs that named rs3001371 ,rs2494738 ,rs1130214 were selected from 3 representative BLOCK Districts of PKB1.Two independent samples t test was used to analysis P300 and CNV between two groups,and the same way to analysis the average level of P300 and CNV and PKB1 SNP genolatency of P2(P＜0.05) and lower amplitude of P3a(P＜0.01 ) ,P3b(P＜0.01 ) and P3 (P＜0.01 ) ;CNV had der had statistical difference (P＜ 0.05 )in PKB1 rs3001371 gene between C/C and C/T genotype combined which included C allele, and T/T genotype.The amplitude of P3a( (5.93 ± 2.35 ) μV, P3b(6.51 ± 3.00) μV, P3 (6.27±2.43) μV) were lower than TT Genotype ( (7.45 ±2.19)μV, (8.63 ±3.57)μV,(8.04 ±2.57)μV,respectively).The mean of CNV indicators were not found different in statistics among the rs3001371 genotypes.Conclusions PKB1 gene rs3001371 polymorphism is associated with the principal component of P300 amplitude in patients with Major depressive disorder which suggest that genetic factors may have a certain impact on cognitive function in the patients with Major depressive disorder.

Objective To analyze the survival rate of HIV /AIDS patients receiving highly active antiretroviral therapy(HAART)since the implementation of the national Four Free and One Carepolicy against HIV in Hangzhou.Methods Clinical data of 2370 AIDS patients were collected from National AIDS Comprehensive Treatment Information System Treatment Library from 2004 to 2014.The data, including basic information,viral load,CD4 +T lymphocyte counts,starting time of treatment,WHO clinical stage,infection pathways and follow-up were respectively analyzed.Kaplan-Meier and Cox proportional hazards models were used to analyze the survival rate and the factors affecting survival.Results The total follow-up time was 3968.14 person years and 57 patients died in 2370 patients with a mortality rate of 1 .44 /100 person years (57 /3968.14).Kaplan-Meier method showed that the cumulative survival rates of the first,third and fifth year were 98.08%,96.20% and 95.24%,respectively.The overall mortality rate fell from 6.06 /100 person years in 2006 to 1 .44 /100 person years in 2014.The mortality rate of AIDS-related disease declined from 1 .10 /100 person years in 2009 to 0.90 /100 person years in 2014.Multivariate Cox regression analysis showed that the risk of death for patients with CD4 +T 200-349 cells/μL was 0.466 times(95%CI 0.246-0.882)as that for patients with CD4 +T cells <200 /μL.The risk of death was 3.408 times(95%CI 1 .365-8.506)in patients aged≥ 50 years,3.788 times(95%CI 1 .645-8.718)in patients aged 40 to <50 years,and 2.593 times(95%CI 1 .139-5.905)in patients aged 30 to 40 years as that in patients aged <30 years.The mortality risk for patients with baseline WHO stage Ⅲ and Ⅳ was 1 .960 times as patients with WHO stage Ⅰ and Ⅱ (95% CI 1 .117-3.439 ).Conclusions Patients with increased age,low CD4 +T counts and baseline WHO stage Ⅲ or Ⅳ are main risk factors affecting survival rate of HIV /AIDS patients,early antiviral therapy is the key for improving the survival rate of patients.

Objective : To investigate the HIV-1 molecular transmission network among school students with newly confirmed HIV/AIDS in Hangzhou City from 2020 to 2021, so as to provide insights into AIDS control in school. Methods : School students with newly confirmed HIV/AIDS in Hangzhou City from 2020 to 2021 were sampled, and participants' demographics and epidemiological survey data were retrieved from the HIV/AIDS Control Information System of Chinese Disease Prevention and Control Information System. The HIV-1 pol gene was amplified in participants' blood samples using reverse transcription (RT) and nested PCR assays, and the HIV subtypes were characterized through multiple sequence alignments. The gene sequences were aligned among newly confirmed HIV/AIDS patients in Hangzhou City during the same study period. The genetic distance was estimated using the software MEGA 11, and the molecular transmission network was created using the software Cytoscape 3.9.1 to analyze the characteristics of clustered cases in the network. Results: A total of 99 school students were newly confirmed with HIV/AIDS in Hangzhou City from 2020 to 2021, including 98 men (98.98%), and 94 men who had sex with men (94.94%). The HIV-1 pol gene sequence was successfully amplified from 87 cases, and four HIV-1 subtypes were characterized, including CRF07_BC (49 cases, 56.32%), CRF01_AE (27 cases, 31.03%), CRF55_01B (6 cases, 6.90%) and CRF67_01B (5 cases, 5.75%). There were 30 molecular clusters in 50 MSM, with 2 to 16 cases in each cluster, and 27 molecular clusters associated with non-student cases, and clustered cases were all MSM. Infection route, location of schools and current address of non-student cases were identified as factors affecting case clustering, and the residence of schools was an important area of activity among clustered HIV/AIDS cases. Conclusions CRF07_BC and CRF01_AE were predominant HIV-1 subtypes among school students with newly confirmed HIV/AIDS in Hangzhou City from 2020 to 2021, and the transmission of HIV/AIDS strongly correlated with non-student cases, with men-men sexual behaviors as the predominant transmission route. The interventions for of MSM is recommended to be reinforced to block the transmission of HIV/AIDS from community to schools.