OBJECTIVE: To explore the genotype-phenotype correlation in a Chinese family with structural brain abnormalities due to variant of the TUBB gene. METHODS: A family undergoing prenatal diagnosis at Beijing Obstetrics and Gynecology Hospital in October 2024 was selected as the study subject. Clinical data were collected. Amniotic fluid sample was subjected to chromosomal copy number variation sequencing (CNV-seq). Trio whole-exome sequencing (Trio-WES) was carried out on the amniotic fluid and parental blood samples, and candidate variant was verified by Sanger sequencing. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: 2023-KY-076-01). RESULTS: Both prenatal ultrasound and fetal MRI showed deviation of brain midline, unilateral lateral ventriculomegaly, and bilateral gyral asymmetry. Trio-WES revealed that the fetus has harbored a maternally derived heterozygous missense variant of the TUBB gene [NM_178014.4: c.155A>G (p.N52S)]. Sanger sequencing confirmed that the woman and a previously terminated fetus both harbored the same variant. Both the proband and two fetuses exhibited similar neuroimaging abnormalities including midline deviation and asymmetrical gyri. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PM2_Supporting+PS2_Moderate+PS3). CONCLUSION The heterozygous c.155A>G (p.N52S) variant was the TUBB gene probably underlay the pathogenesis of the structural brain abnormalities in this family. Above findings have expanded the phenotypic spectrum associated with the variant and facilitated the prenatal diagnosis for this family.
Humans ; Female ; Pregnancy ; Prenatal Diagnosis ; Tubulin/genetics* ; Adult ; Brain/diagnostic imaging* ; Male ; Pedigree ; DNA Copy Number Variations/genetics* ; Exome Sequencing ; Genetic Association Studies ; Magnetic Resonance Imaging

ObjectiveTo find out whether there is any difference in the monitoring results of mosq-ovitraps placed in different orientations in multi-storey residential areas, so as to provide a scientific basis for routine and emergency monitoring of Aedes albopictus with mosq-ovitraps in residential areas. MethodsFrom July 6th to October 26th 2023, one mosquito ovitrap was set up in each of the 4 orientations of east, south, west and north around the buildings in a multi-storey residential area in Jinhui Town, Fengxian District, Shanghai. Data was collected and recorded 72 hours after placement. The chi-square test was used to compare the mosquito ovitrap indices (MOIs) of two independent samples, and the Kruskal⁃Wallis H test was used to compare the MOIs of multiple independent samples. ResultsAfter 16 weeks of surveillance, 997 mosquito ovitraps were recovered, of which 211 were positive, with the mosquito ovitrap index (MOI) of 21.16% and the Aedes albopictus density index of 1.03 mosquitoes·ovitrap^-1. The MOIs were higher in September (24.22%) and October (23.96%), and the MOIs in the west, south and north within the two months were all above 20.00%. From July to October, the MOIs in the east, west, south and north were 20.70%, 22.20%, 25.50% and 16.20%, respectively, and the difference in MOIs among the 4 orientations was not statistically significant (χ²=6.647, P=0.084). Stratified analysis by month showed that in August, the south side of the multi-storey residential areas had the highest MOI (31.30%), the north side had the lowest MOI (1.30%), and there was a statistically significant difference in MOI in the east, west, south and north (χ²=25.986, P<0.001). In October, the MOI in the west was the highest (33.30%) and the MOI in the east was the lowest (6.30%), the difference in MOIs of the 4 orientations was statistically significant (χ²=12.007, P=0.007). The MOIs in the south side of the building in the outskirts of the residential area from the 1st week in July to the 4th week in October was lower (19.20%) than that in the south side of the inner building (31.70%), and the difference in MOI was statistically significant (χ²=5.118, P=0.024). ConclusionThe study of MOI in different orientations in a multi-storey residential area is a preliminary exploration based on field work, and the results show that there is a difference in MOIs in different orientations during the peak breeding period of mosquitoes. Further indicators such as temperature, humidity and wind speed in different orientations can be collected to explore the influencing factors of MOIs.

Objective:To evaluate the long-term protective efficacy of the recombinant Chinese hamster ovary cell-derived hepatitis B vaccine（CHO-HepB）26 years post-vaccination in the rural China.Methods:Zhengding county，Hebei province was designated as a rural monitoring site for CHO-HepB efficacy. Study participants included individuals born between 1997 and 1999 who had completed the three-dose CHO-HepB primary series without booster doses. A cross-sectional survey was conducted in late 2024 using random sampling. Demographic and vaccination history data were collected via questionnaires，and hepatitis B virus（HBV）serological markers were detected using chemiluminescence. Historical surveillance data were integrated to infer infection statuses of HBsAg-positive individuals and evaluate longitudinal trends in anti-HBs seropositivity and antibody titers.Results:Among 178 participants（mean time since vaccination：26.2 years），the seroprevalence rates were 0.6% for HBsAg（95% CI：0.0%-1.6%），64.6% for anti-HBs（95% CI：57.6%-71.6%），and 1.1% for anti-HBc（95% CI：0.0%-2.7%）. Compared to the pre-vaccination baseline HBsAg positivity of 11.3% in children under 10 years of age，the estimated vaccine protection rate was 95%. Two notable cases were identified：one with concurrent HBsAg and anti-HBc positivity and one with anti-HBs and anti-HBc positivity，suggestive of transient HBV exposure（1999—2009）without chronicity. Natural immune boosting was inferred for the latter case based on anti-HBs titer dynamics. Longitudinal analysis of four prior cross-sectional surveys（2005，2009，2013，and 2017）revealed no significant upward trends in HBsAg and anti-HBc positivity（both P>0.05）over 26 years，while anti-HBs seropositivity declined significantly（ P<0.05）from 6 to 26 years post-vaccination. Conclusion:The CHO-HepB vaccine demonstrates sustained immunological persistence and robust long-term protection up to 26 years post-immunization. Continued emphasis on rigorous implementation of mother-to-child transmission prevention strategies is critical for future hepatitis B control.

Objective:To evaluate the effects of laser moxibustion(LM)on tumor-related immunity in tumor-bearing mice.Methods:Thirty male C57BL/6 mice were randomized into a control group,a model group,and an LM group,with 10 mice in each group.Mice in the control group did not receive any intervention,and mice in the model group and LM group were injected into the right armpit with 1×107 cells/mL Lewis lung carcinoma cells to induce tumor models.From the first day after the injection,the LM group mice were irradiated with LM at Guanyuan(CV4)and bilateral Zusanli(ST36),5 min each point for a total of 15 min,once daily for 15 consecutive days.From the day of visible tumor formation(i.e.,day 8),mice in the model and LM groups were measured every other day with a vernier caliper to calculate the tumor volume.On day 16 after injection,the serum,lung,spleen,and tumors of the mice were harvested.Hematoxylin-eosin staining was used to observe tumor pulmonary metastasis.Changes in the serum levels of programmed death 1(PD-1)and programmed death-ligand 1(PD-L1)were analyzed by enzyme-linked immunosorbent assay;the abundance of PD-1 and PD-L1 proteins in tumors was determined by Western blotting;and the proportions of lymphocyte subsets and myeloid-derived suppressor cells(MDSCs)in tumors and spleen were determined by flow cytometry.Results:Compared to the model group,the tumor volume of mice in the LM group decreased significantly on days 8 and 10(P<0.05);the lung tissues of the LM group showed no apparent tumor atypia;the proportion of monocytic MDSCs(M-MDSCs)in the tumors of LM group mice decreased(P<0.05);the PD-1 and PD-L1 levels in the serum of mice in the LM group declined with statistically significant variation in the reduction of PD-L1(P<0.01);PD-1 and PD-L1 protein expression in the tumor tissues of mice in the LM group reduced significantly(P<0.01).Conclusion:LM intervention can reduce the expression of PD-1 and PD-L1 in the serum of tumor-bearing mice,decrease the expression of PD-1 and PD-L1 proteins and the proportion of M-MDSCs in tumor tissues.These effects may be one of the mechanisms by which LM slows the early-stage growth of Lewis lung carcinoma tumor in mice.

Purpose This study aimed to investigate the expression of CTLA-4 and PD-L1 in pulmonary lympho-epithelial carcinoma(PLEC)and to explore their relationships with patient prognosis and with tumor-infiltrating lym-phocytes(TILs).Methods Fifty cases of PLEC were retrospectively collected,together with 23 samples of adjacent normal lung tissue.Immunohistochemistry was performed to detect CTLA-4 and PD-L1 expression in both PLEC and adjacent normal lung tissues,as well as to quantify CD4+and CD8+T-lymphocytes infiltration within the tumor micro-environment.CTLA-4,PD-L1,and the distributions of CD4+T cells and CD8+T cells were then correlated with the clinicopathological features of PLEC.Results The positive rate of CTLA-4 in PLEC was significantly higher than that in adjacent normal lung tissue(P＜0.05).PD-L1 expression differed significantly across TNM stages of PLEC(P＜0.05)and was positively correlated with TNM stages(r=0.31,P=0.03).CD4+and CD8+T-lymphocytes were pre-dominantly localized in the tumor stroma,with CD4+T cells density exceeding that of CD8+(P＜0.05).Within canc-er nests,CD8+T cells density was significantly higher than CD4+(P＜0.05).Conclusion Both PD-L1 and CTLA-4 are frequently expressed in PLEC,suggesting they represent potential immunotherapeutic targets.In the PLEC micro-environment,lymphocytes primarily infiltrated the stromal compartment,and CD4+T cells are more abundant than CD8+T cells in that locale.

Objectives:To evaluate the effectiveness of functionalized decellularized extracellular matrix(F-dECM)prepared from porcine auricular cartilage or porcine rib cartilage in repairing xenogeneic articular osteochondral defects.Methods:The por-cine auricular cartilage and rib cartilage were crushed and decellularized.Heparin modification and multi-factor loading were a-chieved through amide bond reaction.The physical characteristics of the particles were characterized by SEM observation and mer-cury intrusion.The chemical characteristics of the particles were characterized by tissue section staining and kit detection.The rabbit knee joint full-thickness defect models were established and implanted by different particles respectively.After 6 weeks and 12 weeks of operation,the samples were taken for gross observation,CT scanning and tissue sectioning to comprehensively evaluate the repair effect.Results:After heparin modification,the glycosaminoglycan lost during the decellularization process was replen-ished.Both heparinized auricular cartilage and costal cartilage presented a highly developed porous structure.Among them,the porosity and pore diameter of costal cartilage were both lower than those of auricular cartilage(P＜0.05).During the observation periods of 6 weeks and 12 weeks,the porcine costal cartilage F-dECM implantation group was superior to other experimental groups in knee joint repair effects(P＜0.05).Conclusion:Costal-derived F-dECM is a joint osteochondral defect repair material with application potential.

Objective:To evaluate the effects of laser moxibustion(LM)on tumor-related immunity in tumor-bearing mice.Methods:Thirty male C57BL/6 mice were randomized into a control group,a model group,and an LM group,with 10 mice in each group.Mice in the control group did not receive any intervention,and mice in the model group and LM group were injected into the right armpit with 1×107 cells/mL Lewis lung carcinoma cells to induce tumor models.From the first day after the injection,the LM group mice were irradiated with LM at Guanyuan(CV4)and bilateral Zusanli(ST36),5 min each point for a total of 15 min,once daily for 15 consecutive days.From the day of visible tumor formation(i.e.,day 8),mice in the model and LM groups were measured every other day with a vernier caliper to calculate the tumor volume.On day 16 after injection,the serum,lung,spleen,and tumors of the mice were harvested.Hematoxylin-eosin staining was used to observe tumor pulmonary metastasis.Changes in the serum levels of programmed death 1(PD-1)and programmed death-ligand 1(PD-L1)were analyzed by enzyme-linked immunosorbent assay;the abundance of PD-1 and PD-L1 proteins in tumors was determined by Western blotting;and the proportions of lymphocyte subsets and myeloid-derived suppressor cells(MDSCs)in tumors and spleen were determined by flow cytometry.Results:Compared to the model group,the tumor volume of mice in the LM group decreased significantly on days 8 and 10(P<0.05);the lung tissues of the LM group showed no apparent tumor atypia;the proportion of monocytic MDSCs(M-MDSCs)in the tumors of LM group mice decreased(P<0.05);the PD-1 and PD-L1 levels in the serum of mice in the LM group declined with statistically significant variation in the reduction of PD-L1(P<0.01);PD-1 and PD-L1 protein expression in the tumor tissues of mice in the LM group reduced significantly(P<0.01).Conclusion:LM intervention can reduce the expression of PD-1 and PD-L1 in the serum of tumor-bearing mice,decrease the expression of PD-1 and PD-L1 proteins and the proportion of M-MDSCs in tumor tissues.These effects may be one of the mechanisms by which LM slows the early-stage growth of Lewis lung carcinoma tumor in mice.

Objectives:To evaluate the effectiveness of functionalized decellularized extracellular matrix(F-dECM)prepared from porcine auricular cartilage or porcine rib cartilage in repairing xenogeneic articular osteochondral defects.Methods:The por-cine auricular cartilage and rib cartilage were crushed and decellularized.Heparin modification and multi-factor loading were a-chieved through amide bond reaction.The physical characteristics of the particles were characterized by SEM observation and mer-cury intrusion.The chemical characteristics of the particles were characterized by tissue section staining and kit detection.The rabbit knee joint full-thickness defect models were established and implanted by different particles respectively.After 6 weeks and 12 weeks of operation,the samples were taken for gross observation,CT scanning and tissue sectioning to comprehensively evaluate the repair effect.Results:After heparin modification,the glycosaminoglycan lost during the decellularization process was replen-ished.Both heparinized auricular cartilage and costal cartilage presented a highly developed porous structure.Among them,the porosity and pore diameter of costal cartilage were both lower than those of auricular cartilage(P＜0.05).During the observation periods of 6 weeks and 12 weeks,the porcine costal cartilage F-dECM implantation group was superior to other experimental groups in knee joint repair effects(P＜0.05).Conclusion:Costal-derived F-dECM is a joint osteochondral defect repair material with application potential.

Purpose This study aimed to investigate the expression of CTLA-4 and PD-L1 in pulmonary lympho-epithelial carcinoma(PLEC)and to explore their relationships with patient prognosis and with tumor-infiltrating lym-phocytes(TILs).Methods Fifty cases of PLEC were retrospectively collected,together with 23 samples of adjacent normal lung tissue.Immunohistochemistry was performed to detect CTLA-4 and PD-L1 expression in both PLEC and adjacent normal lung tissues,as well as to quantify CD4+and CD8+T-lymphocytes infiltration within the tumor micro-environment.CTLA-4,PD-L1,and the distributions of CD4+T cells and CD8+T cells were then correlated with the clinicopathological features of PLEC.Results The positive rate of CTLA-4 in PLEC was significantly higher than that in adjacent normal lung tissue(P＜0.05).PD-L1 expression differed significantly across TNM stages of PLEC(P＜0.05)and was positively correlated with TNM stages(r=0.31,P=0.03).CD4+and CD8+T-lymphocytes were pre-dominantly localized in the tumor stroma,with CD4+T cells density exceeding that of CD8+(P＜0.05).Within canc-er nests,CD8+T cells density was significantly higher than CD4+(P＜0.05).Conclusion Both PD-L1 and CTLA-4 are frequently expressed in PLEC,suggesting they represent potential immunotherapeutic targets.In the PLEC micro-environment,lymphocytes primarily infiltrated the stromal compartment,and CD4+T cells are more abundant than CD8+T cells in that locale.

Objective:To evaluate the long-term protective efficacy of the recombinant Chinese hamster ovary cell-derived hepatitis B vaccine（CHO-HepB）26 years post-vaccination in the rural China.Methods:Zhengding county，Hebei province was designated as a rural monitoring site for CHO-HepB efficacy. Study participants included individuals born between 1997 and 1999 who had completed the three-dose CHO-HepB primary series without booster doses. A cross-sectional survey was conducted in late 2024 using random sampling. Demographic and vaccination history data were collected via questionnaires，and hepatitis B virus（HBV）serological markers were detected using chemiluminescence. Historical surveillance data were integrated to infer infection statuses of HBsAg-positive individuals and evaluate longitudinal trends in anti-HBs seropositivity and antibody titers.Results:Among 178 participants（mean time since vaccination：26.2 years），the seroprevalence rates were 0.6% for HBsAg（95% CI：0.0%-1.6%），64.6% for anti-HBs（95% CI：57.6%-71.6%），and 1.1% for anti-HBc（95% CI：0.0%-2.7%）. Compared to the pre-vaccination baseline HBsAg positivity of 11.3% in children under 10 years of age，the estimated vaccine protection rate was 95%. Two notable cases were identified：one with concurrent HBsAg and anti-HBc positivity and one with anti-HBs and anti-HBc positivity，suggestive of transient HBV exposure（1999—2009）without chronicity. Natural immune boosting was inferred for the latter case based on anti-HBs titer dynamics. Longitudinal analysis of four prior cross-sectional surveys（2005，2009，2013，and 2017）revealed no significant upward trends in HBsAg and anti-HBc positivity（both P>0.05）over 26 years，while anti-HBs seropositivity declined significantly（ P<0.05）from 6 to 26 years post-vaccination. Conclusion:The CHO-HepB vaccine demonstrates sustained immunological persistence and robust long-term protection up to 26 years post-immunization. Continued emphasis on rigorous implementation of mother-to-child transmission prevention strategies is critical for future hepatitis B control.