Objective To perform laboratory detection and molecular traceability analysis on a case of imported kala-azar in Shenzhen to determine the infection strain. Methods Bone marrow puncture fluid and blood samples from a case of kala-azar in Shenzhen were collected for laboratory tests. The patient's bone marrow puncture fluid smears were stained with Giemsa and examined under a microscope. Blood samples were examined for antibodies using the rk39 visceral leishmania rapid diagnostic reagent. Whole blood DNA was extracted, and the ITS-1 sequence was amplified by PCR, sequenced and aligned, and a phylogenetic tree was constructed based on the ITS-1 sequence. Results Microscopic examination of the patient's bone marrow smears revealed a large number of Leishmania amastigotes without flagella, confirming the diagnosis of kala-azar. The patient's blood was tested positive with the rk39 rapid diagnostic reagent, and PCR amplification yielded an ITS-1 gene product sequence that matched the expected size. Sequence alignment with the NCBI database showed 100% sequence similarity with the ITS-1 gene sequence of Leishmania infantum, confirming the infecting strain as Leishmania infantum. Phylogenetic tree construction of the amplified ITS-1 sequence revealed clustering into a clade with Leishmania infantum , and close to KC347299, one of the reference sequence selected. Conclusions The case of kala-azar in Shenzhen was caused by Leishmania infantum. Kala-azar still occurs in China, so the diagnostic technology of medical personnel in non-epidemic areas should be strengthened so that they can actively use new diagnostic technologies to assist in diagnosis, thus improving their prevention and control ability of Leishmania parasites.

Abstract: Objective　To analyze and understand the mutations of drug resistance genes in imported Plasmodium falciparum in Shenzhen, aiming to assess the efficacy of antimalarial drugs and guide effective drug use. Methods A total of 85 samples from individuals with imported Plasmodium falciparum confirmed by fluorescence quantitative polymerase chain reaction (PCR) in Shenzhen from 2022 to 2023 were collected and genomic DNA was extracted. Nested PCR was used to amplify resistance genes, including Plasmodium falciparum Kelch 13 (PfK13), multidrug resistance gene 1 (Pfmdr1), chloroquine resistance transporter (Pfcrt), dihydrofolate reductase (Pfdhfr), and dihydropteroate synthase (Pfdhps) genes. Bidirectional sequencing was conducted, and mutations in these resistance genes were analyzed using MEGA11.06 software. Results　The study found one missense mutation (S549P) and four synonymous mutations in PfK13. For Pfmdr1, 62.69% of the samples showed Y184F mutation, and no N86Y mutation was detected. No mutations at positions 72 and 73 were detected in the Pfcrt gene, while mutations at M74I, N75E, and K76T accounted for 17.46%, 15.87%, and 15.87%, respectively. The wild-type of Pfcrt gene is dominant (82.54%, 52), followed by the triple mutant I74E75T76 (15.87%, 10). The most common mutation type for Pfdhfr is I51R59N108 (91.78%, 67), followed by the wild type (2.74%, 2). More than half (60.32%, 38) of the Pfdhps samples were wild-type, with single mutation K540E being the most common mutation type. S436A, G437A, K540E, A581G, A613S, I431V, G556K, and G579E site mutations were detected. Among the Pfdhfr-Pfdhps combination mutations, I51R59N108-E540 was the most frequent combination mutation (11.48%), with 59.02% of samples showing solitary Pfdhfr mutations. Conclusions In this study, PfK13 mutation rates were low, with no reported resistance mutations. The Y184F mutation emerged as the dominant Pfmdr1 mutation, with no detection of N86Y. For Pfcrt, the wild-type was dominant, followed by the I74E75T76 triple mutation variant. Triple mutant I51R59N108 of Pfdhfr was very common, and our study did not find Pfdhfr Pfdhps completely resistant and super resistant mutants, but there were other quintuple and septuple mutant types. In the future, it is crucial to continue to strengthen the monitoring of malaria parasite resistance genes and to further integrate in vivo efficacy monitoring to effectively guide clinical drug use.

The Coronavirus disease 2019 (COVID-19) has spread globally. Although mixed liver impairment has been reported in COVID-19 patients, the association of liver injury caused by specific subtype especially chronic hepatitis B (CHB) with COVID-19 has not been elucidated. In this multi-center, retrospective, and observational cohort study, 109 CHB and 327 non-CHB patients with COVID-19 were propensity score matched at an approximate ratio of 3:1 on the basis of age, sex, and comorbidities. Demographic characteristics, laboratory examinations, disease severity, and clinical outcomes were compared. Furthermore, univariable and multivariable logistic and Cox regression models were used to explore the risk factors for disease severity and mortality, respectively. A higher proportion of CHB patients (30 of 109 (27.52%)) developed into severe status than non-CHB patients (17 of 327 (5.20%)). In addition to previously reported liver impairment markers, such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and total bilirubin, we identified several novel risk factors including elevated lactate dehydrogenase (⩾ 245 U/L, hazard ratio (HR) = 8.639, 95% confidence interval (CI) = 2.528-29.523; P < 0.001) and coagulation-related biomarker D-dimer (⩾ 0.5 µg/mL, HR = 4.321, 95% CI = 1.443-12.939; P = 0.009) and decreased albumin (< 35 g/L, HR = 0.131, 95% CI = 0.048-0.361; P < 0.001) and albumin/globulin ratio (< 1.5, HR = 0.123, 95% CI = 0.017-0.918; P = 0.041). In conclusion, COVID-19 patients with CHB were more likely to develop into severe illness and die. The risk factors that we identified may be helpful for early clinical surveillance of critical progression.
COVID-19 ; Cohort Studies ; Hepatitis B, Chronic/epidemiology* ; Humans ; Retrospective Studies ; Risk Factors

Objective: To explore the association between drinking behavior and self injury behavior in adolescents. Methods: A total of 9 247 students from 4 middle schools were investigated. Drinking behavior and self injury behavior were collected from questionnaire survey. Univariate and multivariate Logistic regression analysis were used to analyze the relationship between drinking behavior and self injury behavior. Results: Among the 9 247 middle school students, 52.8% reported ever drinking, 24.9% reported drinking behavior in the past 30 days, and 14.6% reported been drunk in the past year. The average age of drinking for the first time was 12.47±3.05. About 47.2% of the participants had self injury behavior. Male with younger drinking age ( OR =1.52), had been drunken ( OR =1.35) and frequent drinking ( OR =1.54) increased the incidence of self injury. Female reported drinking at younger age ( OR =1.69), had been drunk ( OR =1.82) and lived in cities and towns ( OR =1.20) had a higher risk of self injury. Conclusion Drinking at younger age, heavy and frequent drinking are associated with higher risk of self injury in adolescents in sex specific fashion.

Objective:To test whether the constructed intervention model of advance care planning (ACP) for patients with advanced cancer can be successfully implemented and the preliminary intervention effect, which provides reference for empirical research.Methods:32 cases of advanced cancer patients and 25 cases of their families at the Fifth Affiliated Hospital of Zunyi Medical University, Zhuhai were selected. The patients were subdivided into the experimental group and the control group by random number table method. The control group received routine nursing, while the experimental group adopted the intervention model of "VIP for future care" on the basis of customary nursing. We measured the main outcome indicators: enrollment rate, consent rate, completion rate and loss of follow-up rate and secondary outcome indicators: decision-making certainty, end-of-life care preference and post-intervention satisfaction of patients and their families, within 1 week and 1 month after intervention.Results:The enrollment rate, consent rate and loss of follow-up rate were 74.6% (206/276), 36.9% (76/151)and 15.6% (5/32), respectively. After intervention, the completion rates of the experimental group and the control group were 16/16 and 15/16 within one week, and 14/16 and 13/16 within one month. All of the family members were conducted during the follow-up period. The intervention mode of "VIP for future care" had a statistically significant difference in decision-making certainty between the two groups of patients ( β=0.63, 95% CI 0.08-1.18, P<0.05), no statistically significant difference in end-of-life care preference between the two groups of patients and their families ( P>0.05), and had a statistically significant difference in "whether to recommend this project to others" between the two groups ( χ2 value was 4.167 , P<0.05). Conclusions:On the premise of sufficient preparation, the "VIP for future care" intervention mode can be successfully implemented in advanced cancer patients in mainland China, can improve the decision-making certainty of patients and the satisfaction of patients and their families, and it is recommended. And should be applied to ACP intervention for patients with advanced cancer.

Objective:To investigate the epidemiological characteristics of dengue fever and genotyping of the epidemic strains of dengue virus in Shenzhen City in 2018.Methods:Descriptive epidemiological analysis was used to analyze dengue fever prevalence in Shenzhen City in 2018. Blood samples of patients with dengue fever were collected. The colloidal gold immunochromatography was used to detect serum specific IgM and IgG antibodies, and real-time fluorescence quantitative polymerase chain reaction (PCR) was used to detect viral nucleic acids and to identify genotypes. The E gene sequence of isolated virus strain was amplified by reverse transcription PCR. Homology comparison and phylogenetic tree of dengue fever epidemic strains in different countries and regions were conducted. Results:A total of 234 cases of dengue fever were reported in Shenzhen City from January 1 to December 31, 2018. The incidence rate was 1.87/100 000. There were 144 (61.54%) local patients and 90 (38.46%) imported patients, who were mainly from Southeast Asia and surrounding cities. Two hundred and two (86.32%) cases were reported during the epidemic peak period from August to November of the year. The patients mainly aged 20 to 50 years old (195 cases, 83.33%). Dengue virus type (DENV)-1 accounted for 86.01%(166/193), DENV-2 accounted for 10.36%(20/193), DENV-3 accounted for 2.59%(5/193), and DENV-4 accounted for 1.04%(2/193). The local cases were all infected with DENV-1. The homologies of nucleotide sequence and the deduced amino acid sequence of E gene of 24 DENV-1 strains with HAWAII45 strain were 93.0% to 94.6% and 96.6% to 97.2%, respectively. The phylogenetic tree of DENV-1 strains revealed that 23 strains belonged to genotypeⅠ, and one strain belonged to genotype Ⅳ which was the first reported imported cases in Shenzhen City. The homologies of nucleotide sequence and the deduced amino acid sequence of E gene of six DENV-2 strains with NGC strain were 93.1%to 93.9% and 97.0% to 97.8%, respectively. The phylogenetic tree of DENV-2 strains showed that two strains belonged to genotype Cosmopolitan and four strains belonged to genotype Asian Ⅰ, which were first reported in Shenzhen City. Conclusions:The epidemic of dengue fever in Shenzhen City in 2018 has the characteristics of coexistence of local and imported transmission. The main epidemic genotype is DENV-1. It infers that the major virus strains may be imported from Southeast Asia countries and surrounding cities. Therefore, attention should be paid to the epidemic trend of local dengue fever.

Ischemic stroke is prone to recurrence, which is the leading cause of death and disability in stroke. Using predictive model to assess the recurrence risk in patients with ischemic stroke, stratifying risk, and formulating corresponding interventions to reduce the recurrence rate are the core contents of secondary prevention. In this paper, research progress on the prediction of recurrence risks of ischemic stroke recently is summarized from the predicting content or factors, including clinical factors, imaging factors, biological markers and traditional Chinese Medicine syndrome differentiation, and the methods of constructing prediction models about traditional statistical methods and machine learning, in order to provide a reference for clinical work.

Objective: To investigate the genetic characteristics of VP1 genes carried by coxsackievirus A16 strains isolated from cases of hand foot and mouth disease (HFMD) in Shenzhen during 2016 to 2017. Methods: Fecal and anal swab specimens were collected from patients with mild HFMD in four sentinel hospitals and the Institute of Pathogen Biology, Shenzhen Center for Disease Control and Prevention, China during 2016 to 2017. All specimens were tested for CVA16 viral RNA using real-time RT-PCR. The VP1 genes of 51 randomly selected CVA16 strains were amplified by RT-PCR and then sequenced using TaKaRa Biomedical Technology (Dalian). Bioinformatics software, including Mega6.02, BioEdit and DNAStar, was used for comparison and analysis of the VP1 genes. Results: CVA16 strains in Shenzhen during 2016 to 2017 mainly belonged to B1a and B1b subtypes as well as an emerging subtype B3. The epidemic of B1b subtype was found in both 2016 (28 strains) and 2017 (19 strains), while the B1a subtype (two strains) was only detected in 2017. Two B3 subtype strains were detected in 2017. The strains of B1b subtype were closely related to the strains isolated in Shanghai (JQ314149), Wenzhou (KP289416) and Beijing (KU254598), while the B1a subtype strains were closely related to the strains isolated in Kunming (JQ316639) and Tailand (GQ184139). The B3 subtype strain was an emerging CVA16 epidemic strain in mainland China. Further comparison of the CVA16 epidemic strains in Shenzhen area during 2016 to 2017 with the CVA16 strains causing severe neurological symptoms showed that two amino acid mutations (S14N and M23L) were found in VP1 protein. Conclusions The epidemic strains of CVA16 were B1b subtype in Shenzhen area in 2016. However, B1a, B1b and the emerging B3 subtype strains were prevalent in 2017. Compared with the CVA16 strains causing severe neurological symptoms, the CVA16 strains circulating in Shenzhen during 2016 to 2017 carried two amino acid mutations inVP1 protein.

OBJECTIVE： To establish a method for simultaneously determining the contents of 6 volatile components such as eugenol， methyl eugenol， isoeugenol， α-asarone， alkaloid and myristyl ether in Zhachong shisan. METHODS：Volatile oil was extracted by steam distillation and analyzed by gas chromatography. The chromatographic column was ov-1701， with hydrogen flame ionization detector. The carrier gas was nitrogen， the inlet temperature was 220 ℃， the detector temperature was 240 ℃， the flow rate was 1.0 mL/min， the injection volume was 1 μL， the split ratio was 10 ∶ 1， with programmed temperature. RESULTS： The linear ranges of eugenol， methyl eugenol， isoeugenol， α-asarone， costunolide and myristyl were 2.00-12.00， 1.00-6.00， 0.60-3.60， 0.05-0.30， 0.05-0.30， 0.60-3.60 mg/mL（r＝0.999 9）. The detection limits were 0.01， 0.01， 0.02， 0.01， 0.01， 0.01 mg/mL， respectively； the limits of quantification were 0.05， 0.03， 0.06， 0.02， 0.02， 0.03 mg/mL， respectively. RSDs of precision， stability and reproducible tests were all lower than 3％； average recovery rates were 99.33％-100.04％， RSDs were 0.02％-0.35％ （n＝6）； RSDs of durability test were all lower than 5％. CONCLUSIONS： The established method was simple and reproducible. It can be used for simultaneous determination of the contents of 6 volatile components in Zhachong shisan.

Objective: To understand the awareness level of AIDS knowledge and its influencing factors among middle school students from AIDS high prevalence areas of Yi Autonomous Prefecture (Liangshan Prefecture), and to provide a reference for making propaganda strategies of AIDS knowledge and intervention measures in the relevant departments. Methods: Totally 10 749 students in 4 middle schools were selected from AIDS high prevalence areas of Yi Autonomous Prefecture in Liangshan by the method of cluster sampling. A self-made questionnaire was used to investigate the ways of getting AIDS knowledge and the condition of conducting AIDS life training skills in school. Results: The awareness rate of AIDS-related knowledge among middle school students in Liangshan Prefecture was 44.4%. According to the results of multivariate Logistic regression analysis, female students, junior grade, Yi nationality, poor self-evaluation, lack of parents, living in rural areas, good family economic condition (student self-assessment), living in a community of Yi nationality were all the dangerous factors to the awareness rate of AIDS. School education, radio and television, and health promotion were the main ways for students to acquire AIDS knowledge(88.82%, 80.83%, 73.54%). Minority students, students in severely affected areas received less AIDS life skills training in schools, the time was later, and student needs were not strong. Conclusion The AIDS awareness level of middle school students in Liangshan AIDS high-incidence area is too low, so it is necessary to pay attention to develop targeted programs to strengthen AIDS health education for middle school students.