ObjectiveTo genotype the specimens from hand-foot-and-mouth disease (HFMD) cases in Dehong Prefecture, Yunnan Province in 2023 by gene sequencing, and to analyze the genetic characteristics of Coxsackievirus A4 (CVA4), CVA6, CVA10, CVA16 and enterovirus A71 (EV-A71), so as to clarify the serotypes of enteroviruses causing endemic HFMD and the genotype composition of CVA4, CVA6, CVA10, CVA16 and EV-A71 prevalent in HFMD, and to provide baseline genotype data for monitoring the dynamic changes of the five virus genotypes. MethodsStool specimens from HFMD cases in Dehong Prefecture in 2023 that tested positive for EV-A71, CVA16 and other enteroviruses by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) were first subjected to gene sequencing and typing of the VP4/VP2 junction region to determine the viral types. Subsequently, the complete VP1 gene of each virus was amplified in two segments using group-specific VP1 gene primers corresponding to enterovirus groups (i.e., enterovirus group A, B, and C) and sequenced (group A virus: primers 486/488 and 487/489, group B virus: primers 490/492 and 491/493, group C virus: primers 494/496 and 495/497). The representative reference sequences of VP1 genes for CVA4, CVA6, CVA10, CVA16 and EV-A71 were downloaded from the literature, and the VP1 gene phylogenetic tree was constructed using MEGA 6.0 software for genetic characterization. ResultsA total of 256 stool specimens were collected from HFMD cases in Dehong Prefecture in 2023, of which 94 tested positive for nucleic acid by RT-qPCR, yielding a positive rate of 36.72%. Of the 94 positive specimens, 83 were identified as group A viruses, and were classified into 5 serotypes (17 strains of CVA4, 3 of CVA6, 6 of CVA10, 49 of CVA16, and 8 of EV-A71). Nine strains were group B viruses, comprising 2 serotypes: 2 strains of echovirus type 3 (E3) and 7 strains of Coxsackievirus B type 5 (CVB5). The remaining 2 strains were group C viruses, both identified as poliovirus type 3 (PV3). Genetic analyses of the VP1 region showed that all 17 CVA4 strains were identified as subgenotype C2, all 3 CVA6 strains as subgenotype D3a, all 6 CVA10 strains as genotype C. Among the 49 CVA16 strains, 46 were identified as subgenotype B1a and 3 as subgenotype B1b, and all 8 EV-A71 strains were identified as subgenotype C4a. ConclusionIn 2023, group A viruses dominated the pathogenic spectrum of HFMD in Dehong Prefecture. Among them, CVA16 had the highest positivity rate. Genetic characteristic analyses showed that CVA4, CVA6, CVA10 and EV-A71 were each composed of a single genotype (subgenotype), while CVA16 was composed of two subgenotypes (B1a and B1b), with B1a being the predominant one. It is recommended that molecular epidemiological surveillance of HFMD pathogens, particularly group A viruses, be continuously strengthened in Dehong Prefecture.

[This corrects the article DOI: 10.1016/j.apsb.2021.08.008.].

Objective:To analyze the non-enterovirus A71 (non-EVA71) and non-coxsackievirus A16 (non-CVA16) enteroviruses causing hand, foot and mouth disease (HFMD) in Kunming and Qujing of Yunnan Province in 2021 by sequencing the VP4/VP2 and VP1 genes and to analyze the phylogenetic characteristics of the VP1 gene of CVA2, aiming to provide reference for the prevention and control of CVA2.Methods:The samples were made and extracted strictly according to the Laboratory Manual for Hand, Foot and Mouth Disease (China Center for Disease Control and Prevention, 2018 Edition). VP4/VP2 junction regions were firstly amplified and sequenced by MD91/OL68-1 primers. These sequences were firstly edited and then "blasted" on the GenBank to determine the virus serotype. To analyze the phylogenetic characteristics of CVA2, the entire VP1 gene sequences were amplified in two segments using enterovirus species A primers. Virus serotype was again confirmed online by "Enterovirus Genotyping Tool Version 0.1". The sequences of the reference virus genotypes/sub-genotypes were downloaded according to the reference. The phylogenetic trees were constructed by Mega5.2 software and the genetic characteristics were analyzed.Results:A total of 749 non-EVA71 and non-CVA16 enteroviruses were detected in the two areas in 2021. Group A enteroviruses were the main pathogens, with CVA16 as the predominant virus, and a small number of group B enteroviruses were reported. Only five strains of CVA2 were detected with a detection rate of 0.67% (5/749), indicating that CVA2 was a rare pathogen for HFMD in the two areas. The sequencing and serotyping results were consistent using the two genomic regions of VP4/VP2 junction region and VP1 region. Phylogenetic analysis showed that three Kunming strains belonged to genotype A, while two Qujing strains belonged to genotype D.Conclusions:The detection rate of CVA2 in Kunming and Qujing was 0.67% in 2021. CVA2 was a rare pathogen for HFMD in the two regions. Phylogenetic analysis showed genotypes A and D spread in Kunming and Qujing, respectively, but had not caused epidemics. To our knowledge, this was the first report of genotype A of CVA2 in China. Strengthening the laboratory surveillance especially molecular epidemiological surveillance is valuable for the monitor and analysis of transmission source for CVA2.

Objective:To analyze the etiological distribution and phylogenetic characteristics of hand, foot and mouth disease (HFMD) in Qujing city of Yunnan Province in 2020.Methods:Stool samples were collected from HFMD cases in Qujing city in 2020 and virus RNA was extracted directly from treated stool suspensions. Virus VP4 gene sequences were firstly amplified using MD91/OL68-1 primer pairs and sequenced, then the virus serotypes were determined by BLAST search on the GenBank. Virus entire VP1 gene sequences were amplified and sequenced. Virus serotypes were identified online using Enterovirus Genotyping Tool Version 0.1. Sequences of reference virus genotypes/sub-genotypes were downloaded according to references. Phylogenetic trees were constructed by MEGA5.2 software and the genetic characteristics were analyzed.Results:A total of 47 strains of enteroviruses (EVs) were detected with a detection rate of 10.22% (47/460). The detected viruses were coxsackievirus A4 (CVA4, 0.65%, 3/460), CVA6 (7.83%, 36/460), CVA10 (0.87%, 4/460) and CVA16 (0.87%, 4/460). All were enterovirus species A (EVA), while other group viruses were not detected. The predominant virus was CVA6, accounting for 7.83% (36/460). EVA71 was not detected. CVA4 strains of C2 and C4 subgenotypes were co-circulating strains in Qujing city. CVA6 subgenotype D3a and CVA16 subgenotype B1a were also circulated in Qujing city. All CVA10 strains were in a separate lineage.Conclusions:Similar to the previous situation in China, the detection rates of EVA71 and CVA16 were very low, even zero. This study showed that CVA6 was the predominant virus, indicating a HFMD outbreak caused by CVA6 in Qujing city in 2020. The phylogenetic analysis showed CVA10 isolates belonged to a separate lineage, which might be unique to Qujing city. Laboratory and molecular epidemiological surveillance of non-EVA71 and non-CVA16 viruses, especially CVA6 and CVA10 should be strengthened in the future.

Substantial progress in the use of chemo-photodynamic nano-drug delivery systems (nano-DDS) for the treatment of the malignant breast cancer has been achieved. The inability to customize precise nanostructures, however, has limited the therapeutic efficacy of the prepared nano-DDS to date. Here, we report a structurally defined tandem-responsive chemo-photosensitive co-nanoassembly to eliminate primary breast tumor and prevent lung metastasis. This both-in-one co-nanoassembly is prepared by assembling a biocompatible photosensitive derivative (pheophorbide-diphenylalanine peptide, PPA-DA) with a hypoxia-activated camptothecin (CPT) prodrug [(4-nitrophenyl) formate camptothecin, N-CPT]. According to computational simulations, the co-assembly nanostructure is not the classical core-shell type, but consists of many small microphase regions. Upon exposure to a 660 nm laser, PPA-DA induce high levels of ROS production to effectively achieve the apoptosis of normoxic cancer cells. Subsequently, the hypoxia-activated N-CPT and CPT spatially penetrate deep into the hypoxic region of the tumor and suppress hypoxia-induced tumor metastasis. Benefiting from the rational design of the chemo-photodynamic both-in-one nano-DDS, these nanomedicines exhibit a promising potential in the inhibition of difficult-to-treat breast tumor metastasis in patients with breast cancer.

Objective:To analyze the etiological characteristics of hand, foot and mouth disease (HFMD) cases and to investigate the genetic characteristics of VP1 region of coxsackievirus A6 (CVA6) and CVA16 strains in Wenshan prefecture of Yunnan Province in 2020.Methods:Virus RNA was extracted directly from stool samples of children with HFMD in Wenshan prefecture of Yunnan Province in 2020. Enterovirus (EV) VP4/VP2 junction region was amplified using MD91/OL68-1 primers and sequenced, and then the serotypes of EV isolates were preliminarily identified. Amplification and sequencing of the complete VP1 gene were performed. A phylogenetic tree was constructed by MEGA 5.2 software with the reference strains from GenBank. Genetic and molecular epidemiological characteristics of CVA6 and CVA16 were analyzed.Results:Thirty-two strains of EV and one strain of astrovirus MLB1 were detected in 317 specimens with an overall virus detection rate of 10.41% (33/317). Among the 32 EV strains, 31 (96.88%) were enterovirus species A (EVA) and one (3.12%) was EVB. EVC and EVD were not detected. CVA6 was the predominant EV, accounting for 62.50% (20/32), followed by CVA16 (18.75%, 6/32), CVA4 (9.37%, 3/32) and CVA10 (3.12%, 1/32). EVA71 was not detected. The phylogenetic analysis showed 20 CVA6 strains belonged to D3a sub-genotype and could be further divided into three clusters. Six CVA16 strains belonged to B1a sub-genotype, which was one of the predominant genotypes circulating in China, and could be divided into two clusters.Conclusions:The detection rate of pathogens causing HFMD in Wenshan prefecture in 2020 was 10.41% and the most common etiologic agents were CVA6 and CVA16. Based on the genetic analysis of the VP1 gene, the predominant genotype circulating in Wenshan prefecture in 2020 was CVA6 D3a sub-genotype, followed by CVA16 B1a sub-genotype. EVA71 was not detected.

Objective:To explore the clinical application value of group A Streptococcus (GAS) antigen rapid detection method in children suffering from GAS infection.Methods:A total of 44 733 children with suspected GAS infection who were admitted to the Outpatient and Inpatient Departments of Shenzhen Children′s Hospital from January 2015 to December 2019.Throat swab specimens from all children were collected, and BinaxNOW Strep A Test reagent was used for GAS antigen rapid detection.Among them, the throat swabs of 346 children were inoculated with blood culture medium for traditional bacterial culture, and then the GAS antigen rapid detection was tested.The sensitivity and specificity of the two methods were compared, and according to the result of the GAS antigen rapid detection, its age, gender and seasonal trends were analyzed.SPSS 19.0 software was applied for statistical analysis of the data.Results:Among the 346 children tested by both methods, the results of bacterial culture were adopted as the reference method, the sensitivity of the rapid detection method for GAS antigen was 89.41%(152/170 cases), and the specificity was 94.32%(166/176 cases) compared with culture methods.A total of 44 733 cases GAS antigen were tested in children in Shenzhen, of which 10 024 cases were positive, with the positive detection rate of 22.41%.The trend of GAS antigen rapid detection was consistent with the five-year trend, with the high positive rate of 3-8 years, of which 4-6 years of positive rate was the highest.The two seasonal peaks were evident each year, with peaks occurring in April-June, and November and January of next year.The detection rate ratio of male and female was 1.74∶1, and the gender difference was significant ( χ2=27.93, P<0.000 1). GAS antigen rapid detection rate in different clinical departments from high to low in order are as follows: dermatology outpatient (52.34%), emergency clinic (47.74%), internal medicine outpatient (37.36%), infectious disease area (19.71%), five-level disease area (10.27%), internal medicine area (8.63%), surgical areas (7.34%) and neonatal areas (0). Conclusions:GAS antigen rapid detection method and bacterial culture method have high coincidence rate, and high sensitivity and specificity, and can be popularized and applied in the diagnosis of GAS infectious diseases in children.GAS detection rate is higher in outpatient emergency department and dermatology clinics.There are obvious differences from seasonal and population (age and gender) in the positive detection of GAS antigen.No neonates were found.

Objective:To investigate the diagnosis and management of partial or complete hydatidiform mole with coexistent intrauterine pregnancy.Methods:Clinical data of 10 cases of hydatidiform mole with coexistent intrauterine pregnancy admitted to the Third Affiliated Hospital of Guangzhou Medical University, from September 2009 to May 2019 were retrospectively described.Results:(1) During the same period, 65 960 women were delivered at our hospital, and hydatidiform mole with coexistent intrauterine pregnancy was accounted for 1/6 596, among which complete hydatidiform mole and coexisting fetus (CHMCF) and partial hydatidiform mole and coexistent fetus (PHMCF) were found in four and six cases, respectively. The mean age of the ten patients were (30.9±4.1) years old, ranging from 26 to 35 years old, with 2.5 (1-4) times of pregnancies. Nine cases were identified at 22 +3 (12 +3-32 +3) gestational weeks and one at 9 + weeks. (2) Recurrent vaginal bleeding during pregnancy occurred in six cases, nausea and vomiting in three cases, and hyperthyroidism in mid- and late pregnancy in two cases. One patient developed preeclampsia and one case of severe mitral regurgitation with mild pulmonary hypertension. (3) In the 10 patients, the summit serum β -hCG level was 139 935 (16 990-546 033) U/L, and CHMCF and PHMCF patients were 212 500 (200 000-546 033) U/L and 60 768 (16 990-225 000) U/L, respectively. (4) The ultrasound results revealed a dark honeycomb area of the placenta in five cases, placental thickening in two cases, and vesicular placenta in one case. One case was found with bilateral giant luteinized ovarian cyst by ultrasound, multiple metastases in the left lower lobe of the lung by chest CT, multiple nodules in the pleural wall of the left lung by lung MRI, and CHMCF by pelvic MRI. In one case, ultrasound at 14 weeks of gestation showed interrupted fetal abdominal wall, visible mass, gastric bubble, liver, part of the intestinal echoes, and omphalocele. One case was found with embryo arrest. (5) The karyotype analysis of one case through amniocentesis was 46,XX with no anomalies, and chromosome microarray analysis was arr[hg19](1-22)×2. Prenatal diagnosis was refused in the remaining cases. (6) Among the ten patients, three were terminated by rivanol intra-amniotic injection, two received drug abortion, and uterine evacuation, and two with spontaneous abortion followed by curettage with a visible fetus and hydatidiform tissue. Total hysterectomy was performed in one patient due to partial invasion of the uterus by hydatidiform mole. One patient underwent a cesarean section on account of the left lower lung metastasis. One case developed preeclampsia at 33 +4 weeks of gestation and delivered two premature infants by cesarean section. Pathology examination found a complete and partial vesicular fetal mass in four and six cases, with P57 (－) and P57 (+), respectively. (7) During the follow-up, two women developed the persistent trophoblastic disease and received chemotherapy, while the remaining eight cases did not. Conclusions:When hydatidiform mole with coexistent intrauterine pregnancy is found, a timely differential diagnosis between CHMCF and PHMCF is needed. CHMCF is at a higher risk of abortion, intrauterine death, premature delivery, preeclampsia, and other maternal complications. Therefore, termination of CHMCF should be individualized. Most PHMCF patients have fetal malformation or fetal loss; thereby, timely termination is recommended.

Objective To detect the expression levels of peripheral blood CXCL10 and its receptor CXCR3 and T cell subsets in of the patients with advanced vitiligo and the influence of compound Chinese medicine on it.Methods Flow cytometry was used to detect the cellular proportions of peripheral blood T cell subsets,ELISA was employed to quantify serum CXCL10 and CXCR3 expression levels before and after treatment.Results After 1 month of taking Chinese medicine,the proportions of CD3+ CD4+ cells and CD3+ CD8+ cells were increased compared before treatment(P＜0.05).The expression level of peripheral serum CXCL10 before treatment was significantly increased compare with the healthy control group(P＜0.01),and the CXCL10 level after treatment was decreased significantly compared with that before treatment(P＜0.05).The expression level of peripheral serum CXCR3 was significantly increased compared with the healthy control group(P＜0.05),while which after treatment was still significantly higher than that in the healthy control group(P＜0.05).Conclusion CXCL10,CXCR3 and T cell subsets proportion may be involved in the pathogenesis of vitiligo.The compound Chinese medicine used in this study plays the curative effect possibly by regulating T cell subsets and expression levels of CXCL10 and CXCR3.

Objectives To study the clinical effects of percutaneous kyphoplasty on senile osteoporotic vertebral body compression fractures.Methods The 87 elderly patients with osteoporotic vertebral body compression fractures were randomly grouped into control group (n=42) treated with percutaneous vertebroplasty and observation group (n=45) treated with percutaneous kyphoplasty in our hospital from June 2013-March 2016.And the clinical curative effect before and after treatment was compared and analyzed.Results The visual analog scale (VAS) pain score in observation group versus control group was (8.49±2.86) vs.(8.56±2.98),(t=0.11,P>0.05) before treatment,and[(2.58±1.34) vs.(3.34±2.01),t=2.09,P< 0.05],[(2.06±0.97) vs (3.87±1.96),t=5.51,P<0.05]and[(1.09±0.89) vs (2.37±1.29)],t=5.42,P<0.05]at 1 week,4 weeks,and 12 weeks after treatment respectively,showing statistically significant improvement after treatment.The data of kyphosis angle by using Cobb angle,Oswestry lumbar dysfunction index (ODI),mean time of postoperative ambulation,and mean postoperative hospital stay were similar to that of the visual analog scale (VAS) pain score,showing statistically significant improvement after treatment (all P<0.05).The incidence[cases (%)]of bone cement leakage was lower in observation group[3(6.7)]than in control[9(21.4)](χ2=3.98,P<0.05).Conclusions Clinical treatment effects of percutaneous kyphoplasty on the elderly patients with osteoporotic vertebral body compression fractures are superior to that of percutaneous vertebroplasty.Percutaneous kyphoplasty can effectively relieve pain,and improve body function disorders and the quality of life.