Objective:To compare the efficacy of the detection kit based on MicroDrop microdroplet digital PCR platform that can identify mycoplasma pneumonia and common drug-resistance mutation,and throughout targeted next-generation sequencing(tNGS)in detecting common drug-resistance mutations of mycoplasma pneumonia.Methods:A total of 300 samples of clinical respiratory tract of pneumonia inpatients at Guangdong Provincial People's Hospital between 2023 and 2024 were collected.Both the detection kit for drug-resistance mutation of mycoplasma pneumoniae and the tNGS method were employed to detect drug-resistance mutation genes.For samples with inconsistent results,Sanger sequencing was used for verification.Results:For the 300 samples,the detection rates of positive mycoplasma pneumonia of the detection kit for drug-resistance mutation of mycoplasma pneumoniae and the tNGS method were respectively 87.00%and 78.67%,with a Kappa value of 0.711,indicating a relatively high level of agreement between the two methods.Among 25 samples with inconsistent results,Sanger sequencing was employed for validation.The results revealed that for samples with low-frequency gene mutations,the reagent kit maintained reliable detection capability,whereas tNGS exhibited missed detections.Thus,the reagent kit demonstrates superior performance in detecting low-frequency mutation samples.Conclusion:The detection rate of low-frequency mutation samples by the digital PCR-based mycoplasma pneumoniae drug-resistance mutation detection kit is higher than that of the tNGS method.This approach helps enhance the accuracy of detection results,providing a rapid and precise means of detecting drug-resistance genes for clinical diagnosis and treatment.

Objective:To compare the efficacy of the detection kit based on MicroDrop microdroplet digital PCR platform that can identify mycoplasma pneumonia and common drug-resistance mutation,and throughout targeted next-generation sequencing(tNGS)in detecting common drug-resistance mutations of mycoplasma pneumonia.Methods:A total of 300 samples of clinical respiratory tract of pneumonia inpatients at Guangdong Provincial People's Hospital between 2023 and 2024 were collected.Both the detection kit for drug-resistance mutation of mycoplasma pneumoniae and the tNGS method were employed to detect drug-resistance mutation genes.For samples with inconsistent results,Sanger sequencing was used for verification.Results:For the 300 samples,the detection rates of positive mycoplasma pneumonia of the detection kit for drug-resistance mutation of mycoplasma pneumoniae and the tNGS method were respectively 87.00%and 78.67%,with a Kappa value of 0.711,indicating a relatively high level of agreement between the two methods.Among 25 samples with inconsistent results,Sanger sequencing was employed for validation.The results revealed that for samples with low-frequency gene mutations,the reagent kit maintained reliable detection capability,whereas tNGS exhibited missed detections.Thus,the reagent kit demonstrates superior performance in detecting low-frequency mutation samples.Conclusion:The detection rate of low-frequency mutation samples by the digital PCR-based mycoplasma pneumoniae drug-resistance mutation detection kit is higher than that of the tNGS method.This approach helps enhance the accuracy of detection results,providing a rapid and precise means of detecting drug-resistance genes for clinical diagnosis and treatment.

The changes of carotid intima media thickness (cIMT) in type 2 diabetic patients with vitamin D insufficiency and subclinical macrovascular disease after 12 months' intervention therapy with calcitriol were observed.86 patients with vitamin D insufficiency [25 (OH) D＜30 ng/ml] and subclinical macrovascular disease were randomized to group A (n =44,treated with 0.5 μg/day calcitriol) or group B (n =42,blank control) for 12 months.40 patients in each group completed the follow up.After 12 months of therapy with calcitriol (0.5 μg.qd.po.),levels of high sensitive-Creactive protein,sCD36,and cIMT were significantly decreased,while 25 (OH) D and ankle brachial index were significantly increased (P＜0.05) in group A,suggesting that calcitriol could improve cIMT in patients with vitamin D insufficiency and subclinical macrovascular disease.

Objective To evaluate the relationship between CD4+ T lymphocyte count and results of enzyme-linked immunospot (ELISPOT) assay in human immunodeficiency virus (HIV)-Mycobacterium tuberculosis (M.tb) coinfected patients.Methods A total of 193 HIV-infected individuals in Yunnan Province and Shanghai were enrolled.T-SPOT.TB assay was employed to detect M.tb specific T lymphocyte in the peripheral blood mononuclear cells (PBMC).CD4+ T lymphocyte in PBMC from the enrolled subjects was detected by flow cytometry.Data were analyzed using t test.ResultsThe incidence of latent tuberculosis in HIV-infected individuals was 30.6％.The CD4+ T lymphocyte counts in HIV-infected individuals with active tuberculosis were 190×106/L,which were significantly lower than those in HIV-infected individuals with latent tuberculosis (484×106/L; t=6.665,P＜0.01).The HIV-infected individuals were stratified according to CD4+ T lymphocyte counts of ＞500×106/L,200×106-500×106/L,and ＜200×106/L and the constituent ratios of active tuberculosis/latent tuberculosis were 1∶16.2,1∶1.3 and 5.6∶1,respectively.Among 79 subjects with positive T-SPOT.TB results,20 were coinfected with active tuberculosis,in which 14 had CD4+ T lymphocyte counts of ＜200 ×106/L,5 had 200×105-500×106/L and 1 had ＞500×106/L.Fifty-two in 59 HIV/latent tuberculosis patients individuals had CD4+ T lymphocyte counts of ＞200×106/L.ConclusionsThe prevalence of latent tuberculosis in HIV-infected individuals is high in China.Cellular immunity in HIV-infected individuals with active tuberculosis is severely impaired.With the decrease of CD4 ′ T lymphocyte counts,patients with latent tuberculosis are prone to develop active tuberculosis in HIV-infected individuals.The negative predictive value of T-SPOT.TB is significantly diminished in patient with low CD4+ T lymphocyte counts,especially less than 200×106/L.

Objective To analyze the situation of inquirer in Shanghai Jing'an district AIDS voluntary coun-seling and testing(VCT). Methods Inquirer were serviced by trained counselors before and after HIV antibody tes-ring,each inquirer were asked to complete a "Advisory Cases Registration Form". Results Inquirer were male-domi-nated,accounting for 85.7%. And inquirer's age were mainly 25 ～ 34 years old, which accounting for 55.8%. The types of inquirer were mainly because of male non-married heterosexual sexual contact, female non-married heterosex-ual contact, male homosexual sexual sexual contact, which accounted for 77.4%, 11.8% and 3.7%, respectively. For the total of three categories accounted for 92.9%. Inquirer aged 25 ～ 34 years old in 2007 were more than 2006 ,and inquirer because of male non-married heterosexual sexual contact in 2007 were more than 2006. There were 2 infectors because of male homosexual sexual contact in 3 positive infectors who were screened in 2007. Conclusion VCT played more and more important role in AIDS prevention and control, and it should be further strengthened and ex-panded.

Aim: To establish a linear additive model for the predication of in vitro sinomenine hydrochloride release from the combination of immediate release, enteric-coated and sustained-release pellets based on the release profiles of each pellet type. Methods: Immediate release pellets were manufactured by extrusion/spher-onization technology. The operation of bottom-spraying in the fluid-bed equipment was conducted to enteric-coating using Eudragit~(R) L-30D-55 and sustained-release coating using Surelease~(R) . In vitro sinomenine hydrochloride release profiles of both uncoated and coated pellets were fitted to the chosen mathematical equations offered by the curve fitting toolbox of Matlab~(R) before a linear additive model was created based upon the best-to-fitting equations. The proportion of each pellet type in the combined format to generate the desired 24 h sinomenine hydrochloride release profile was solved by Matlab~(R). The predicted and assayed sinomenine hydrochloride release from the polled pellets was compared. Results: It was shown that the actual sinomenine hydrochloride release profiles of each pellet type were approximate to those of predicted ones. A linear additive model of the appropriate mathematical equations of each pellet was proven to be capable of controlling in vitro release of sinomenine hydrochloride multiple-unit pellets. Conclusion: A multiple-unit combined system of the selected pellets, as a novel sustained-release system, was successfully prepared. In vitro release performance of the calculated combination of each pellet type could be guaranteed by this approach in designing sustained-release drug delivery system.