BACKGROUND:Reducing the rate of abnormal fertilization is an effective approach to improving the efficacy of in vitro fertilization and reducing patients'financial strain.However,the current research on abnormal fertilization has focused on exploring the types of prokaryotic nuclei and their generation mechanisms,as well as analyzing embryos formed by abnormal fertilization,chromosomal ploidy and utilization value.There is a lack of clinical prediction models for abnormal fertilization based on retrospective studies. OBJECTIVE:To construct a nomogram model to predict abnormal female factors in in vitro fertilization. METHODS:A total of 5 075 patients undergoing treatment for conventional in vitro fertilization at Nanxishan Hospital of Guangxi Zhuang Autonomous Region from March 2017 to March 2022 were retrospectively analyzed.The male confounders were calibrated on a 1:1 propensity score with a match tolerance of 0.02,and 1 672 cases were successfully matched.According to the Vienna Consensus,patients with≥60%normal fertilization capacity were included in the normal fertilization group(n=836)and those with<60%normal fertilization capacity were included in the abnormal fertilization group(n=836).The model and validation groups were obtained by random sampling at a ratio of 7:3.Factors related to the occurrence of abnormal fertilization following conventional in vitro fertilization in the model group were screened using univariate analysis and the best matching factors were selected using the Least Absolute Shrinkage and Selection Operator(LASSO)and included in a multifactorial forward stepwise Logistic regression to identify their independent influencing factors and plot a nomogram.Finally,the prediction model was validated for discrimination,accuracy and clinical application efficacy using receiver operating characteristic curves,calibration curves,clinical decision curves and clinical impact curves. RESULTS AND CONCLUSION:The univariate analysis indicated the factors influencing the occurrence of abnormal fertilization were age,controlled ovarian hyperstimulation protocol,number of assisted pregnancies,years of infertility,infertility factors,anti-mullerian hormone,sinus follicle count,basal luteinizing hormone,luteinizing hormone concentration on the human chorionic gonadotropin day,and estradiol level on human chorionic gonadotropin injection day(P<0.05).LASSO regression further identified the best matching factors,including age,microstimulation protocol,number of assisted pregnancies,years of infertility,anti-mullerian hormone,luteinizing hormone level on human chorionic gonadotropin injection day,and estradiol level on human chorionic gonadotropin injection day(P<0.05).Multifactorial forward stepwise Logistic regression results showed that age,microstimulation protocol,number of assisted conceptions,years of infertility,anti-mullerian hormone,and estradiol level on human chorionic gonadotropin injection day were independent influencing factors for the occurrence of abnormal fertilization following conventional in vitro fertilization.The receiver operating characteristic curves showed an area under the curve of 0.761(0.746,0.777)for the model group and 0.767(0.733,0.801)for the validation group,indicating that the model has good discrimination.The mean absolute error of the calibration curve was 0.044,and the Hosmer-Lemeshow test indicated that there was no significant difference between the predicted probability of abnormal fertilization and the actual probability of abnormal fertilization(P>0.05),indicating the prediction model has good consistency and accuracy.The clinical decision curves and clinical impact curves showed that the model and validation groups had the maximum net clinical benefit at valve probability values of 0.00-0.52 and 0.00-0.48,respectively,and there was a good clinical application efficacy in this valve probability range.To conclude,the nomogram model has good discrimination and accuracy as well as clinical application efficacy for predicting the occurrence of abnormal fertilization in women undergoing conventional in vitro fertilization based on age,microstimulation protocol,number of assisted conceptions,years of infertility,anti-mullerian hormone,and estradiol level on human chorionic gonadotropin injection day.

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.

IL-33 is a member of the IL-1 cytokine superfamily.It is a key regulator of pathological inflammation,immune ho-meostasis and fibrosis.IL-33 receptor ST2 is expressed on the surface of all innate immune cells,as well as some subtypes of B and T cells.IL-33 is a dual-function protein.Under normal circumstances,the N-terminal part of IL-33 resides in the nucleus of its ex-pressed cells and is released as a cytokine to exert immunomodulatory properties when cells are damaged or necrotic.Different expres-sion forms of IL-33 are located differently and may play different immunological roles.The immunological effects of IL-33 are highly di-verse.The cytokine IL-33 plays an amplifying and enhancing role in the innate immune response,while the full-length(FL)IL-33 stored in the nucleus inhibits inflammation by binding to NF-κB.IL-33/ST2 signaling plays a key role in both innate and acquired im-mune responses.In innate immunity,IL-33 and group 2 innate lymphocytes(ILC2)provide an important axis for rapid immune re-sponse and tissue homeostasis.In acquired immunity,IL-33 interacts with dendritic cells,Th2 cells,follicular helper T cells(Tfh)and regulatory T cells(Tregs).IL-33/ST2 signaling triggers pro-tumor or anti-tumor immune responses in different cell types in auto-crine and paracrine ways.In addition,the interaction between IL-33 and mitochondrial metabolism is also one of the potential mecha-nisms affecting the immune system.

Objective To observe the ultrasonic manifestations and outcome of fetuses with absent middle phalanx of the little finger.Methods Data of 36 fetuses with absence of middle segment of the little finger displayed by the first prenatal ultrasound at 16-33 weeks of gestation were retrospectively analyzed.The prenatal ultrasonic manifestations,complicated abnormalities,the chromosomal karyotype results and the outcome of fetuses were recorded.Results Prenatal ultrasound showed 21 fetuses with bilateral absences of middle segment of the little fingers simultaneously with one or more other abnormalities,including 16 fetuses with cardiac malformations,8 with central nervous system malformations,5 with facial malformations,3 with abdominal anomalies and 6 fetuses each complicated with 1 other abnormality.Four of these 21 fetuses received chromosomal karyotype examination,including 2 were found with abnormal chromosomal number(1 of 21-trisomy syndrome and 1 of 18-trisomy syndrome)while the other 2 without chromosomal abnormality.Thereafter,induced labor was performed in 19 cases,while hands DR 3 months after birth displayed ossification centers of middle segment of bilateral little fingers in the rest 2 cases.Among other 15 fetuses prenatal ultrasound showed unilateral(n=2)or bilateral(n=13)absence of middle segment of the little fingers but without other abnormality and underwent chromosomal examination,induced labor was performed in 3 cases with fetal 21-trisomy syndrome,while the other 12 fetuses without abnormal findings were born with no flexion of the little fingers on physical examination,and hands DR 3 months after birth displayed ossification centers of middle segment of bilateral little fingers.Conclusion Displaying of fetal middle segment of the little fingers on prenatal ultrasound was affected by multiple factors,in addition,the ossification might be delayed,and might complicated with other systemic and chromosome abnormalities.Dynamically follow-up with prenatal ultrasound and comprehensively evaluation combined with clinical data was necessary for diagnosing fetal absent middle phalanx of the little finger.

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.

Aging is the process spontaneously occurred in living organisms. Cardiac fibrosis is a pathophysiological process of cardiac aging. Mangiferin is a wellknown C-glucoside xanthone in mango leaves with lots of beneficial properties. In this study, rat model of cardiac fibrosis was induced by injected with 150 mg/kg/d Dgalactose for 8 weeks. The age-related cardiac decline was estimated by detecting the relative weight of heart, the serum levels of cardiac injury indicators and the expression of hypertrophic biomakers. Cardiac oxidative stress and local inflammation were measured by detecting the levels of malondialdehyde, enzymatic antioxidant status and proinflammatory cytokines. Cardiac fibrosis was evaluated by observing collagen deposition via masson and sirius red staining, as well as by examining the expression of extracellular matrix proteins via Western blot analysis. The cardiac activity of profibrotic TGF-β1/p38/MK2 signaling pathway was assessed by measuring the expression of TGF-β1 and the phosphorylation levels of p38 and MK2. It was observed that mangiferin ameliorated D-galactose-induced cardiac aging, attenuated cardiac oxidative stress, inflammation and fibrosis, as well as inhibited the activation of TGF-β1/p38/MK2 signaling pathway. These results showed that mangiferin could ameliorate cardiac fibrosis in D-galactose-induced aging rats possibly via inhibiting TGF-β/p38/MK2 signaling pathway.

Aging is the process spontaneously occurred in living organisms. Cardiac fibrosis is a pathophysiological process of cardiac aging. Mangiferin is a wellknown C-glucoside xanthone in mango leaves with lots of beneficial properties. In this study, rat model of cardiac fibrosis was induced by injected with 150 mg/kg/d Dgalactose for 8 weeks. The age-related cardiac decline was estimated by detecting the relative weight of heart, the serum levels of cardiac injury indicators and the expression of hypertrophic biomakers. Cardiac oxidative stress and local inflammation were measured by detecting the levels of malondialdehyde, enzymatic antioxidant status and proinflammatory cytokines. Cardiac fibrosis was evaluated by observing collagen deposition via masson and sirius red staining, as well as by examining the expression of extracellular matrix proteins via Western blot analysis. The cardiac activity of profibrotic TGF-β1/p38/MK2 signaling pathway was assessed by measuring the expression of TGF-β1 and the phosphorylation levels of p38 and MK2. It was observed that mangiferin ameliorated D-galactose-induced cardiac aging, attenuated cardiac oxidative stress, inflammation and fibrosis, as well as inhibited the activation of TGF-β1/p38/MK2 signaling pathway. These results showed that mangiferin could ameliorate cardiac fibrosis in D-galactose-induced aging rats possibly via inhibiting TGF-β/p38/MK2 signaling pathway.

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.