Cystic echinococcosis, a zoonotic disease that poses a significant threat to human health and animal husbandry development, is prevalent across the world and predominantly occurs in agricultural and pastoral regions. However, cystic echinococcosis cases are rare in non-endemic areas, which is likely to cause misdiagnosis or missing diagnosis, resulting in delay in treatment. This report presents an overseas imported cystic echinococcosis case misdiagnosed as pulmonary and hepatic cysts, so as to provide insights into diagnosis and treatment of cystic echinococcosis in non-endemic areas.

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.

Objective To establish a real-time quantitative polymerase chain reaction (qPCR) assay for B-cell lymphoblastic leukemia according to individualized and specific immunoglobulin gene rearrangements in leukemia cells, and to use it for the monitoring of minimal residual disease (MRD) of B-cell lymphocytic leukemia. Methods The immunoglobulin gene rearrangements of bone marrow samples from 15 cases of B-cell lymphoblastic leukemia were analyzed with a validated European BIOMED-2 system, and the individualized and specific qPCR-based quantification of leukemic immunoglobulin gene rearrangements was established. Results Unique and specific gene rearrangements of immunoglobulin light and heavy chains were identified in 14 cases and Ig-qPCR based on these gene rearrangements had a sensitivity of 10-5 and high specificity which met the international criteria in 10 patients. Leukemia MRD quantification with immunoglobulin gene rearrangement-based qPCR was similar as compared with other MRD detection methods. Conclusion Immunoglobulin gene rearrangement-based leukemia MRD quantification is feasible, sensitive, specific, precise and much valuable for clinical decision of treatments in B-cell lymphoblastic leukemia.

Objective To compare the clinical outcome of intramedullary nail with locking plate in the treatment of two-part proximal humerus fractures.Methods PubMed,SpringerLink,EMBASE,The Cochrane Library,Medline,Science Direct,CNKI,Wanfang Data and VIP database were searched for relevant studies comparing intramedullary nailing and locking plate fixation of two-part proximal humerus fractures.A meta-analysis of the two methods was conducted using the RevMan 5.0.Differences between the two methods were compared with respect to operation time,intraoperative bleeding,bone union time,Constant score and postoperative complication incidence.Results Seven studies were included in the meta-analysis,with 136 patients treated with intramedullary nail and 159 patients treated with locking plate.Intramedullary nailing showed advantages over locking plate fixation with regards to operation time (WMD =-27.71,95％ CI-36.07--19.35,P ＜ 0.05),intraoperative blood loss (WMD =-114.18,95％ CI-169.91--58.45,P ＜ 0.01) and bone union time (WMD =-2.91,95％ CI-5.80--0.01,P ＜ 0.05).While the two methods showed no significant differences in Constant score (WMD =-2.84,95 ％ CI-5.90--0.22,P ＞ 0.05) and postoperative complication incidence (OR =0.89,95％ CI 0.43-1.84,P ＞ 0.05).Conclusion Intramedullary nail is superior to locking plate in operation time,intraoperative blood loss and bone union time for the treatment of two-part proximal humerus fractures,but the two methods are similar in Constant score and postoperative complications rate.

Objective To identify the species of Biomphalaria snails collected in Shenzhen reservoir,based on the mitochondrial 16S rDNA sequences.Methods The 16S rDNA fragments were amplified by PCR from the genome DNA of Biomphalaria snails,and inserted in plasmid pMD-18T for sequencing.The sequence of 16S rDNA fragment and its phylogenetic relationships with those of other species of Biomphalaria snails were analyzed with BLAST and MEGA4 software.Results The amplified 16S rDNA fragment of the Biomphalaria snails was about 466 bp in length.As aligned with the corresponding sequences of the related Biomphalaria species,the identity of nucleotides was 99％ with 1 isolate of Biomphaltria straminea (B.straminea),98％ with 3 isolates of B.kuhniana,95％ with 1 isolate of B.intermedia,and 94％ with 1 isolate of B.edisoni.Based on the 16S rDNA sequence,the results of phylogenetic analysis with neighbor-joining (NJ) and unweighted pair-group method with arithmetic means (UPGMA) indicated that the snails had close genetic relationships with the B.straminea isolate (Genbank accession NO.AY030213.1) Conclusion The Biomphalaria snails collected in Shenzhen reservoir could be classified as B.straminea based on the characteristics of 16S rDNA sequence.

Objective To discuss the feasibility of preoperative diet by measuring gastric emptying time of carbohydrate and protein nutrient solutions in healthy volunteers.Methods A total of 20 healthy volunteers were collected from August 2013 to May 2014.On the morning of the trial,baseline gastric residual volume of each volunteer was measured with magnetic resonance imaging at 8 a.m.,then each of the 20 healthy volunteers took 12.5％ carbohydrate solution 400 ml (containing 40 g of maltodextrin and 10 g of sucrose) or 12.5％ whey protein solution (containing 50 g whey protein) in 5 minutes.Magnetic resonance imaging was conducted to measure the gastric residual volume every 25 minutes.The volunteers were shifted to the other nutrient solution after a 1-week interval.The gastric emptying time of both nutrient solutions was calculated to generate the curves illustrating the process of gastric emptying.Results The baseline gastric residual volume of the volunteers was (14.90 ± 9.39) ml.The total gastric emptying time of carbohydrate solution was (104.90 ± 27.98) min (95 ％ CI 98.64-111.16 min),while that of whey protein solution was (199.6 ± 34.17) min (95％ CI 184.47-214.73 min).There was a significant difference between these two types of nutrient solution in terms of gastric emptying time (P ＜ 0.000 1).Conclusions The induction of anesthesia could be performed 2 hours after carbohydrate administration,and at least 4 hours after whey protein administration.

Objective To identify the species classification of an ornamental Planorbidae from a flower market in Shanghai and analyze its potential distribution in China. Methods In August 2013,six freshwater snail specimens were collected from the Wanshang flower market. The species was identified by morphology and molecular biology. An ecological niche model was constructed based on the native geographic presence occurrence data,and projected onto the whole of China to predict the poten?tial distribution. Results Their shell external morphology suggested that the specimens belonged to Planorbella trivolvis(Say 1817)of Planorbidae,which is native in North America. The sequence data of a fragment of the mitochondrial cytochrome c oxi?dase subunit I(COI)confirmed its identification. A total of 2 294 georeferenced occurrence points in North America were car?ried out from the Global Biodiversity Information Facility databases and 614 records with coordinates were used to produce a North American native niche model by a maximum entropy method(Maxent). The projection on China results suggested high probabilities of occurrence mostly in Henan Province and its borderland with nearby provinces. Conclusions P. trivolvis is sim?ilarly with Biomphalaria species from shell morphology. It is the first records of the species in China,and the field dispersal is not clear.

Objective To detect infection of Angiostrongylus cantonensis and examine effection of treatment to prepare monoclonal antibodies(McAbs). Methods Six-week-old BALB/c mice were imrnunized by the intraperitoneal injection of e/s antigens of Angiostrongylus cantonensis. Fusion of splecn cells from immunized mice with prepared SP2/0-Ag14 myeloma cells was performed in RPMI 1640. Fused cells were suspended in RPMI 1640 containing 1% HAT and 20% fetal calf serum and dispensed into 96-well cell culture plates. The supernatants of clones were screened by ELISA with sera of patients of angiostrongyliasis.Distribution of cohere antigen of 12D5 and 21B7 monoclonal antibodies was analyzed with immunohistochemistry. Two McAbs ( 12D5 and 21B7) were applied to detect the circulating antigen (CAg) in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA.Results 12D5 McAb was identified as IgG1 and 21 B7 McAb was IgM. Western blot result showed two McAbs could used to identified 55 × 103 protein of adult worms of A. cantonensis. Cohere antigen of 12D5 and 21B7 monoclonal antibodies were distributed on intestine surface of A. cantonensis. The detection rates of CAg in the sera of infected rats 100% (48/48), the detection rates of CAg in the sera of angiostrongyliasis patients was 100% (32/32). No cross-reaction to sera of patients with other infection of parasites, such as clonochiasis, fasiolopsiasis, ancylostomiasis, trichinosis, anisakiasis as well as schsitosomiasis, and health srea did not reacted with 12D5 and 21B7 McAbs,and detaction rate of antibody of angiostrongyliasis patients only reached 75% (24/32) with antigen of A. cantonensis. Conclusion Cohere antigen of 12D5 and 21B7monoclonal antibodies were antigens of enteric epithelium. Sandwich ELISA with 12D5 and 21B7 McAbs showed high specificity act as detecting CAg of A. cantonensis in sera of infection animal and patients. It is apparent that Sandwich ELISA with 12D5 and 21 B7 is not only rapid and simple without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A. cantonensis.

To identify the gene differentially expressed in female Anopheles anthropophagus and to analyze its gene sequence, this gene amplified by PCR was identified by real-time PCR and its cDNA was then amplified with rapid amplification of cDNA ends (RACE) technology. It was found that the expression ratio of the female differentially expressed gene in female and male mosquitoes was 267.49 according to the formula F=2~(-⊿⊿CT).The size of mRNA of the gene was 364 bp, and the amino acid sequence deduced from the open reading frame (ORF) was found to be similar to the sequence of tectin protein of Culex quinquefasciatus and proteins of other species. The mRNA sequence of this gene was submitted to NCBI with a accession number of FJ907236. This gene may provide a foundation for further studies on the biological functions of mosquitoes.