Background::Total human immunodeficiency virus (HIV) DNA and integrated HIV DNA are widely used markers of HIV persistence. Droplet digital polymerase chain reaction (ddPCR) can be used for absolute quantification without needing a standard curve. Here, we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods::The limit of detection, dynamic ranges, sensitivity, and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat (LTR) and human CD3 gene (for total HIV DNA) and ACH-2 cells (for integrated HIV DNA). Forty-two cases on stable suppressive antiretroviral therapy (ART) were assayed in total HIV DNA and integrated HIV DNA. Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive (CD4 +) T-cell counts, CD8 + T-cell counts and CD4/CD8 T-cell ratio, respectively. The assay linear dynamic range and lower limit of detection (LLOD) were also assessed. Results::The assay could detect the presence of HIV-1 copies 100% at concentrations of 6.3 copies/reaction, and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction (95% confidence intervals [CI]: 3.6-6.5 copies/reaction) with linearity over a 5-log 10-unit range in total HIV DNA assay. For the integrated HIV DNA assay, the LLOD was 8.0 copies/reaction (95% CI: 5.8-16.6 copies/reaction) with linearity over a 3-log 10-unit range. Total HIV DNA in CD4 + T cells was positively associated with integrated HIV DNA ( r = 0.76, P <0.0001). Meanwhile, both total HIV DNA and integrated HIV DNA in CD4 + T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8 + T-cell counts. Conclusions::This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity. It can be readily adapted for measuring HIV DNA with non-B clades, and it could be beneficial for testing in clinical trials.

Background::Pathological scars are a disorder that can lead to various cosmetic, psychological, and functional problems, and no effective assessment methods are currently available. Assessment and treatment of pathological scars are based on cutaneous manifestations. A two-photon microscope (TPM) with the potential for real-time non-invasive assessment may help determine the under-surface pathophysiological conditions in vivo. This study used a portable handheld TPM to image epidermal cells and dermal collagen structures in pathological scars and normal skin in vivo to evaluate the effectiveness of treatment in scar patients. Methods::Fifteen patients with pathological scars and three healthy controls were recruited. Imaging was performed using a portable handheld TPM. Five indexes were extracted from two dimensional (2D) and three dimensional (3D) perspectives, including collagen depth, dermo-epidermal junction (DEJ) contour ratio, thickness, orientation, and occupation (proportion of collagen fibers in the field of view) of collagen. Two depth-dependent indexes were computed through the 3D second harmonic generation image and three morphology-related indexes from the 2D images. We assessed index differences between scar and normal skin and changes before and after treatment.Results::Pathological scars and normal skin differed markedly regarding the epidermal morphological structure and the spectral characteristics of collagen fibers. Five indexes were employed to distinguish between normal skin and scar tissue. Statistically significant differences were found in average depth ( t = 9.917, P <0.001), thickness ( t = 4.037, P <0.001), occupation ( t= 2.169, P <0.050), orientation of collagen ( t = 3.669, P <0.001), and the DEJ contour ratio ( t = 5.105, P <0.001). Conclusions::Use of portable handheld TPM can distinguish collagen from skin tissues; thus, it is more suitable for scar imaging than reflectance confocal microscopy. Thus, a TPM may be an auxiliary tool for scar treatment selection and assessing treatment efficacy.

Humans ; Autistic Disorder ; China/epidemiology* ; Cohort Studies ; Autism Spectrum Disorder/genetics* ; Asian People

Objective:To explore the clinical features and survival of newly diagnosed follicular lymphoma (FL) patients with diffuse large B-cell lymphoma (DLBCL) component.Methods:1845 newly diagnosed FL patients aged ≥ 18 years with grades 1-3a in 11 medical centers in China from 2000 to 2020 were included, and patients with DLBCL component were screened. The clinical data and survival data of the patients were retrospectively analyzed, and the prognostic factors were screened by univariate and multivariate analysis.Results:146 patients (7.9% ) with newly diagnosed FL had DLBCL component. The median age was 56 (25-83) years, 79 males (54.1% ) . The pathology of 127 patients showed the proportion of DLBCL component. Patients were divided into two groups according to whether the proportion of DLBCL component was ≥ 50% . The study found that patients with DLBCL component ≥ 50% had higher grade 3 ratio (94.3% vs 91.9% , P=0.010) , Ki-67 index ≥ 70% ratio (58.5% vs 32.9% , P=0.013) and PET-CT SUVmax ≥ 13 ratio (72.4% vs 46.3% , P=0.030) than patients with DLBCL component<50% . All patients received CHOP or CHOP like ± rituximab chemotherapy. The overall response rate (ORR) was 88.2% , and the complete response (CR) rate was 76.4% . In the groups with different proportions of DLBCL component, there was no significant difference in the remission rate after induction treatment and the incidence of disease progression within 2 years after initiation of treatment (POD24) ( P<0.05) . The overall estimated 5-year progression free survival (PFS) rate was 58.9% , and the 5-year overall survival (OS) rate was 90.4% . The 5-year OS rate of POD24 patients was lower than that of non POD24 patients (70.3% vs 98.5% , P<0.001) . Compared with non maintenance treatment of rituximab, maintenance treatment of rituximab could not benefit the 5-year PFS rate (57.7% vs 58.8% , P=0.543) , and the 5-year OS rate had a benefit trend, but the difference was not statistically significant (100% vs 87.8% , P=0.082) . Multivariate analysis showed that failure to reach CR after induction treatment was an independent risk factor for PFS ( P=0.006) , while LDH higher than normal was an independent risk factor for OS ( P=0.031) . Conclusion:FL patients with DLBCL component ≥50% have more invasive clinical and pathological features. CHOP/CHOP like ± rituximab regimen can improve the clinical efficacy of patients. Rituximab maintenance therapy can not benefit the PFS and OS of patients. Failure to reach CR after induction therapy was the independent unfavorable factor for PFS.

Objective:To demonstrate the clinical effect of latissimus dorsi musculocutaneous flap with primary closure in V-Y suture in the repair of major lesions in the anterior chest wall that was left after mastectomies with locally advanced breast cancer (LABC) surgery.Methods:From September 2018 to February 2021, the technique was employed on 14 female cancer patients who had LABC surgery in the Department of Breast Surgery of the First Affiliated Hospital of Kunming Medical University. The patients received radical mastectomies with major resection of cutaneous tegument. The defect areas in chest wall were 15.0 cm×15.0 cm-22.0 cm×35.0 cm. The sizes of flap were 12.0 cm×28.0 cm-18.0 cm×35.0 cm. The sizes of musculocutaneous flap were 12.0 cm×28.0 cm×2.0 cm~18.0 cm×35.0 cm×3.5 cm. All patients were entered the postoperative follow-up through out-patient clinic and telephone interviews.Results:The flap provided an efficient coverage in closing the defects among all 14 patients. Three patients presented small areas (1.0-3.0 cm) of superficial necrosis in Y-cross area of the flap. None of the patient had back swelling. The average operation time was 6.3 hours. Postoperative follow-up varied from 4 to 41 months(18 months in average). The colour, texture, elasticity of the flaps were acceptable, with good shapes. Function of upper limbs was normal in 13 cases without lymphedema, except 1 who had lymphedema of affected limb at 3 years after surgery. Eleven cases had radiotherapy after surgery with good tolerance. None of the cases had local recurrence of breast cancer. Five cases had metastasis, 3 cases died of metastasis.Conclusion:The latissimus dorsi musculocutaneous flap with primary closure in V-Y suture is easy to perform and an reliable and efficient technique in repairing large defects in the anterior chest wall left after a LABC surgery.

Chromosome microarray analysis (CMA) is a cost-effective molecular cytogenetic technique that has been used as a first-line diagnostic test in neurodevelopmental disorders in the USA since 2011. The impact of CMA results on clinical practice in China is not yet well studied, so we aimed to better evaluate this phenomenon. We analyzed the CMA results from 434 patients in our clinic, and characterized their molecular diagnoses, clinical features, and follow-up clinical actions based on these results. The overall diagnostic yield for our patients was 13.6% (59 out of 434). This gave a detection rate of 14.7% for developmental delay/intellectual disability (DD/ID, 38/259) and 12% for autism spectrum disorders (ASDs, 21/175). Thirty-three recurrent (n ≥ 2) variants were found, distributed at six chromosomal loci involving known chromosome syndromes (such as DiGeorge, Williams Beuren, and Angelman/Prader-Willi syndromes). The spectrum of positive copy number variants in our study was comparable to that reported in Caucasian populations, but with specific characteristics. Parental origin tests indicated an effect involving a significant maternal transmission bias to sons. The majority of patients with positive results (94.9%) had benefits, allowing earlier diagnosis (36/59), prioritized full clinical management (28/59), medication changes (7/59), a changed prognosis (30/59), and prenatal genetic counseling (15/59). Our results provide information on de novo mutations in Chinese children with DD/ID and/or ASDs. Our data showed that microarray testing provides immediate clinical utility for patients. It is expected that the personalized medical care of children with developmental disabilities will lead to improved outcomes in long-term developmental potential. We advocate using the diagnostic yield of clinically actionable results to evaluate CMA as it provides information of both clinical validity and clinical utility.
Age Factors ; Child ; Child, Preschool ; China ; epidemiology ; ethnology ; Chromosome Disorders ; genetics ; physiopathology ; Chromosomes ; genetics ; DNA Copy Number Variations ; genetics ; Disease Management ; Female ; Humans ; Infant ; Male ; Microarray Analysis ; methods ; Neurodevelopmental Disorders ; diagnosis ; ethnology ; genetics ; physiopathology

Objective: To investigate the genetic characteristics of Lamivudine-resistant mutation patterns and HBV S gene mutants in patients with chronic hepatitis disease of different disease progression. Methods: Blood samples of LAM-resistant patients with chronic hepatitis disease were collected. HBV RT gene nucleotide sequences were obtained, and then differences in drug-resistant mutation patterns, drug susceptibility and HBV S gene mutants characteristics between the two groups were analyzed. Results: Forty-seven chronic hepatitis B (CHB) patients and 16 HBV-related liver cirrhosis (LC)/HBV-related hepatocellular carcinoma (HCC) patients were included in this study. M204I single point mutation and L180M+ M204I/V were the most common pattern during patients with chronic hepatitis disease (35/63, 55.56%). The numbers of resistant to three nucleos(t)ide analogues in LC/HCC group was higher than CHB group’s (62.50% vs 34.04%, P=0.046). In HBV S gene, more immune associated HBsAg-escape mutations were detected in LC/HCC group than that in CHB group (62.50% vs 31.91%, P=0.031). I126T/V and G145A (for LCC/HCC group, 60%), I126S/T and S117T (for CHB group, 46.67%) were showed as the most common form for HBsAg escape mutations in the two groups. The two groups both detected RT mutations concomitantly with stop codon mutations in S gene (rtA181T/sW172* and rtM204I/sW196*). Conclusions Different characteristics in Lamivudine-resistant mutations and associated HBV S gene mutants were found in patients with chronic hepatitis disease of different disease progression, and LC/HCC patients exhibit more multi-drug resistant variants and immune associated HBsAg-escape mutants than CHB patients.

Objective To investigate the relation between the human leucocytes antigen-DQA1(HLA-DQA1) alleles poly-morphism and the Uygur Han nationality hepatitis B patients in Xinjiang and the genetic susceptibility healthy controls ,which is provide some important clues to seek the susceptible genes and disease-resistant genes of HBV infection for Uygur and Han nation-ality hepatitis B patients .Methods HLA-DQAl alleles of 182 cases of the Hepatitis B patients and 163 people were compared with HBV DNA and ALT level .HLA-DQA1 * 0102 ,-DQA1 * 0104 ,-DQA1 * 0201 ,-DQA1 * 0301 ,-DQA1 * 0302 ,-DQA1 * 0501 genes frequency are detected with PCR-SSP .Results Compared the Uygur hepatitis B patients ALT abnormal and HBV DNA high copy quantity group with healthy controls group in allele′s frequency analysis found that HLA-DQA1 * 0301 ,-DQA1 * 0501 genes had statistic significance(P< 0 .05) .Compared the Han nationality hepatitis B patients ALT abnormal and HBV DNA high copy quanti-ty group with healthy controls group in allele′s frequency analysis ,It was found that HLA-DQA1 * 0102 ,-DQA1 * 0201 ,-DQA1 *0301 genes difference had statistic significance(P < 0 .05) .HLA-DQA1 * 0102 had the statistic significance between high and low copy quantity groups(P< 0 .05) .Compared the Han nationality hepatitis B patients ALT normal and low copy quantity group with healthy controls group in allele′s frequency analysis ,It was found that HLA-DQA1 * 0201 genes had statistic significance(P <0 .05) .HLA-DQA1 * 0102 ,-DQA1 * 0301 had statistic significance between the Han and Uygur nationality for HBV patients(P<0 .05) ;HLA-DQA1 * 0201 ,-DQA1 * 0501 had statistic significance between the Han and Uygur nationality for healthy people(P<0 .05) .Conclusion HLA-DQA1 * 0501 is the protected gene of Uygur hepatitis B patients ;-DQA1 * 0301 is the susceptibility gene .The Han nationality hepatitis B patients group HLA-DQA1 * 0102 ,-DQA1 * 0301 ,-DQA1 * 0302 is the susceptibility genes and -DQA1 * 0201 is the Antagonism gene .

Obesity has become a worldwide public health concern. In particular, obesity caused by excessive energy intake was widespread which has strong causal relationship with food preference behaviors. In this review, we would briefly talk about characteristics of feeding behaviors inducing obesity, as well as the possible neurophysiological mechanisms involved in food preference formation and development. Meantime, we would introduce some regulation strategies to deal with obesity which mainly based on the current feeding behaviors of the obese.
China ; Feeding Behavior ; Food Preferences ; Health Policy ; Humans ; Obesity