Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

Objective:To investigate the regulatory effect of electroacupuncture on visual cortex plasticity in adult amblyopic mice and its mechanism.Methods:Forty-eight SPF male healthy 3-week-old Kunming mice were randomly divided into a normal control group, a monocular form deprivation (MD) group, and an electroacupuncture intervention group by the random number table method, with 16 mice in each group.Except for the normal control group, mice in the other groups had their right eyelids sutured for two weeks to establish an adult MD amblyopia model.The electroacupuncture intervention group received electroacupuncture stimulation at three acupoints, Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB20) for four weeks at five weeks of age.The subjective visual function (paw probing success rate) of each group of mice was measured at five and nine weeks of age, respectively.At nine weeks of age, the changes in flash-visual evoked potential (F-VEP) of mice in each group was detected.The expression of plasticity related proteins synaptophysin (SYP), synaptic protein 1 (SYN1), neurite overgrowth inhibitor A (Nogo-A), and Nogo receptor (NgR) proteins in the contralateral cortex of the deprived mouse eyes was detected by Western blot.Expression of early growth response gene 1 (Egr-1) in the visual cortex of mice was detected by immunohistochemical staining.This study was approved by the Animal Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-17), and the management and use of animals were in accordance with the Laboratory Animal Management and Use Guide of Hunan Children's Hospital.Results:At nine weeks of age, the success rates of paw probing in the normal control group, MD group, and electroacupuncture intervention group were (71.69±10.60)%, (25.54±10.09)%, and (58.25±8.39)%, respectively, with a statistically significant overall difference ( F=5.987, P=0.006).Among them, the success rate of paw probing was significantly lower in the MD group than in the normal control group, and the electroacupuncture intervention group was significantly higher than in the MD group (both P<0.05).There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=63.710, P<0.001), with lower P2 wave amplitude in the deprived eye of the MD group than in the normal control group and the electroacupuncture intervention group, and the differences were statistically significant (both P<0.001).There were significant differences in the expression levels of SYP and SYN1 proteins in the contralateral cortex of the deprived mouse eyes ( F=5.451, 3.871; both P<0.05).The relative expression levels of SYP and SYN1 proteins were significantly lower in the MD group than in the normal control group and electroacupuncture intervention group (all P<0.05).There were significant differences in levels of Nogo-A and NgR proteins in the contralateral cortex of the deprived mouse eyes ( F=4.188, 3.942, both P<0.05).The relative expression levels of Nogo-A and NgR proteins were significantly higher in the MD group than in the normal control group and the electroacupuncture intervention group (all P<0.05).The immunohistochemical staining results showed that compared with the normal control group, the MD group mice had a decrease in the expression of Egr-1 in the contralateral cortical neurons of the deprived eye, and the brown neuron protrusions were indistinguishable.Compared with the MD group, the electroacupuncture intervention group showed significant positive cell expression in the contralateral cortical area of the deprived eye, but the expression intensity was weaker than that of the normal control group. Conclusions:Electroacupuncture treatment of Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB21) acupoints can reactivate the plasticity of the visual cortex in adult amblyopic mice and improve their visual function.The mechanism may be related to the regulation of Nogo-A/NgR signaling pathway.

Objective:To investigate the effects of insulin-like growth factor 2 (IGF-2) on the expression of postsynaptic density protein 95 (PSD95), synaptophysin-1 (SYN1), and synaptophysin (SYP) in the mouse visual cortex and visual function after monocular deprivation (MD).Methods:Sixty-four SPF male Kunming mice aged 3 weeks were randomly divided into 4 groups: normal control group, MD group, MD+ IGF-2 recombinant protein (MD+ IGF-2) group, and MD+ fluoxetine (FLX) group, with 16 mice in each group.The MD group, MD+ IGF-2 group and MD+ FLX group were treated with right eyelid suturing at the beginning of 3 weeks old and eyelid opening at the end of 5 weeks old.The MD+ IGF-2 group was intraperitoneally injected with IGF-2 recombinant protein during MD.The MD+ FLX group was given fluoxetine via drinking water for 4 weeks after eyelid opening.The normal control group and MD group were injected intraperitoneally with bovine serum albumin every day from 3 to 5 weeks of age.At the end of 5 and 9 weeks of age, subjective visual function was evaluated by fore paw touching ground reflex experiment.At the end of 9 weeks of age, objective visual function was assessed by flash visual evoked potentials.After the mice were sacrificed, the left visual cortex of mice in each group was taken, and the expression of PSD95, SYN1, and SYP was assessed by Western blot.This study was approved by the Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-16). The handling of experimental animals was carried out in accordance with the Guidelines for the Management and Use of Laboratory Animals in Hunan Children's Hospital.Results:At the end of 5 and 9 weeks of age, there were overall significant differences in the success rate of fore paw touching ground among different groups of mice ( F=4.83, 3.36; both P<0.05). At the end of 5 weeks of age, the success rate was lower in MD group and MD+ FLX group than in normal control group, and significantly higher in MD+ IGF-2 group than in MD group, with statistically significant differences (all P<0.05). At the end of 9 weeks of age, the success rate was lower in MD group than in normal control group, and significantly higher in MD+ IGF-2 group and MD+ FLX group than in MD group (all P<0.05). There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=13.99, P<0.01). The P2 wave amplitude was significantly lower in MD group than in normal control group and MD+ IGF-2 group (both P<0.01). There was no significant difference in the P2 wave latency of F-VEP among the four groups of mice ( F=2.83, P=0.07). The relative expression levels of PSD95, SYN1 and SYP proteins were 1.00±0.41, 1.00±0.10 and 1.00±0.27 in normal control group, 0.32±0.27, 0.68±0.20 and 0.56±0.28 in MD group, 0.78±0.32, 0.91±0.18 and 0.94±0.22 in MD+ IGF-2 group, 0.89±0.65, 0.98±0.28 and 0.94±0.47 in MD+ FLX group, respectively.There were significant differences in levels of PSD95, SYN1 and SYP in mice visual cortex among different groups ( F=4.24, 5.32, 3.40; all P<0.05). The expressions of PSD95, SYN1 and SYP proteins in the visual cortex were lower in MD group than in normal control group, and higher in MD+ IGF-2 group than in MD group (all P<0.05). Conclusions:Administration of exogenous IGF-2 to mice that underwent MD during the critical period can maintain visual cortex plasticity and protect the visual function to a certain extent.

Infectious diseases continue to pose a threat to global public health. Successive global shocks caused by emerging and re-emerging pathogens have continuously challenged existing surveillance systems, highlighting the urgent need to build efficient and precise pathogen surveillance networks. Pathogen genomic databases have been developed rapidly in recent two decades, significantly improving the molecular identification, evolutionary analysis, and transmission tracking of pathogens, and changing disease surveillance strategies and patterns. This paper summarizes the developmental history and current state of pathogen genomic databases, and discusses their applications in public health, including pathogen variation surveillance, emerging or suspected pathogen identification, and epidemiological tracing. Furthermore, this paper systematically analyzes the limitations and key challenges faced by current global health prevention and control system, and suggests the focus of the development of online pathogen databases to address existing shortcomings, ultimately improve global infectious disease surveillance and early warning

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

Objective:To investigate the regulatory effect of electroacupuncture on visual cortex plasticity in adult amblyopic mice and its mechanism.Methods:Forty-eight SPF male healthy 3-week-old Kunming mice were randomly divided into a normal control group, a monocular form deprivation (MD) group, and an electroacupuncture intervention group by the random number table method, with 16 mice in each group.Except for the normal control group, mice in the other groups had their right eyelids sutured for two weeks to establish an adult MD amblyopia model.The electroacupuncture intervention group received electroacupuncture stimulation at three acupoints, Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB20) for four weeks at five weeks of age.The subjective visual function (paw probing success rate) of each group of mice was measured at five and nine weeks of age, respectively.At nine weeks of age, the changes in flash-visual evoked potential (F-VEP) of mice in each group was detected.The expression of plasticity related proteins synaptophysin (SYP), synaptic protein 1 (SYN1), neurite overgrowth inhibitor A (Nogo-A), and Nogo receptor (NgR) proteins in the contralateral cortex of the deprived mouse eyes was detected by Western blot.Expression of early growth response gene 1 (Egr-1) in the visual cortex of mice was detected by immunohistochemical staining.This study was approved by the Animal Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-17), and the management and use of animals were in accordance with the Laboratory Animal Management and Use Guide of Hunan Children's Hospital.Results:At nine weeks of age, the success rates of paw probing in the normal control group, MD group, and electroacupuncture intervention group were (71.69±10.60)%, (25.54±10.09)%, and (58.25±8.39)%, respectively, with a statistically significant overall difference ( F=5.987, P=0.006).Among them, the success rate of paw probing was significantly lower in the MD group than in the normal control group, and the electroacupuncture intervention group was significantly higher than in the MD group (both P<0.05).There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=63.710, P<0.001), with lower P2 wave amplitude in the deprived eye of the MD group than in the normal control group and the electroacupuncture intervention group, and the differences were statistically significant (both P<0.001).There were significant differences in the expression levels of SYP and SYN1 proteins in the contralateral cortex of the deprived mouse eyes ( F=5.451, 3.871; both P<0.05).The relative expression levels of SYP and SYN1 proteins were significantly lower in the MD group than in the normal control group and electroacupuncture intervention group (all P<0.05).There were significant differences in levels of Nogo-A and NgR proteins in the contralateral cortex of the deprived mouse eyes ( F=4.188, 3.942, both P<0.05).The relative expression levels of Nogo-A and NgR proteins were significantly higher in the MD group than in the normal control group and the electroacupuncture intervention group (all P<0.05).The immunohistochemical staining results showed that compared with the normal control group, the MD group mice had a decrease in the expression of Egr-1 in the contralateral cortical neurons of the deprived eye, and the brown neuron protrusions were indistinguishable.Compared with the MD group, the electroacupuncture intervention group showed significant positive cell expression in the contralateral cortical area of the deprived eye, but the expression intensity was weaker than that of the normal control group. Conclusions:Electroacupuncture treatment of Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB21) acupoints can reactivate the plasticity of the visual cortex in adult amblyopic mice and improve their visual function.The mechanism may be related to the regulation of Nogo-A/NgR signaling pathway.

Objective:To investigate the effects of insulin-like growth factor 2 (IGF-2) on the expression of postsynaptic density protein 95 (PSD95), synaptophysin-1 (SYN1), and synaptophysin (SYP) in the mouse visual cortex and visual function after monocular deprivation (MD).Methods:Sixty-four SPF male Kunming mice aged 3 weeks were randomly divided into 4 groups: normal control group, MD group, MD+ IGF-2 recombinant protein (MD+ IGF-2) group, and MD+ fluoxetine (FLX) group, with 16 mice in each group.The MD group, MD+ IGF-2 group and MD+ FLX group were treated with right eyelid suturing at the beginning of 3 weeks old and eyelid opening at the end of 5 weeks old.The MD+ IGF-2 group was intraperitoneally injected with IGF-2 recombinant protein during MD.The MD+ FLX group was given fluoxetine via drinking water for 4 weeks after eyelid opening.The normal control group and MD group were injected intraperitoneally with bovine serum albumin every day from 3 to 5 weeks of age.At the end of 5 and 9 weeks of age, subjective visual function was evaluated by fore paw touching ground reflex experiment.At the end of 9 weeks of age, objective visual function was assessed by flash visual evoked potentials.After the mice were sacrificed, the left visual cortex of mice in each group was taken, and the expression of PSD95, SYN1, and SYP was assessed by Western blot.This study was approved by the Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-16). The handling of experimental animals was carried out in accordance with the Guidelines for the Management and Use of Laboratory Animals in Hunan Children's Hospital.Results:At the end of 5 and 9 weeks of age, there were overall significant differences in the success rate of fore paw touching ground among different groups of mice ( F=4.83, 3.36; both P<0.05). At the end of 5 weeks of age, the success rate was lower in MD group and MD+ FLX group than in normal control group, and significantly higher in MD+ IGF-2 group than in MD group, with statistically significant differences (all P<0.05). At the end of 9 weeks of age, the success rate was lower in MD group than in normal control group, and significantly higher in MD+ IGF-2 group and MD+ FLX group than in MD group (all P<0.05). There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=13.99, P<0.01). The P2 wave amplitude was significantly lower in MD group than in normal control group and MD+ IGF-2 group (both P<0.01). There was no significant difference in the P2 wave latency of F-VEP among the four groups of mice ( F=2.83, P=0.07). The relative expression levels of PSD95, SYN1 and SYP proteins were 1.00±0.41, 1.00±0.10 and 1.00±0.27 in normal control group, 0.32±0.27, 0.68±0.20 and 0.56±0.28 in MD group, 0.78±0.32, 0.91±0.18 and 0.94±0.22 in MD+ IGF-2 group, 0.89±0.65, 0.98±0.28 and 0.94±0.47 in MD+ FLX group, respectively.There were significant differences in levels of PSD95, SYN1 and SYP in mice visual cortex among different groups ( F=4.24, 5.32, 3.40; all P<0.05). The expressions of PSD95, SYN1 and SYP proteins in the visual cortex were lower in MD group than in normal control group, and higher in MD+ IGF-2 group than in MD group (all P<0.05). Conclusions:Administration of exogenous IGF-2 to mice that underwent MD during the critical period can maintain visual cortex plasticity and protect the visual function to a certain extent.

Objective This study aimed to investigate the efficacy of modified endoscopic autologous cartilage eustachian tube pharyngeal orifice tuboplasty(MEACETT)in patients with patulous eustachian tube(PET).Meth-ods A retrospective analysis was conducted on the clinical data of 27 patients(30 ears)diagnosed with PET who underwent MEACETT.Autologous cartilage was used through the incision at the posterior end of the inferior turbi-nate and filled into the lateral wall of the pharyngeal orifice of the eustachian tube.Without affecting the movement function of the eustachian tube during swallowing,the collapse of the pharyngeal orifice was fully filled.Before and after the surgery,the visual analogue scale(VAS),the eustachian tube dysfunction questionnaire-7(ETDQ-7)and hospital anxiety and depression scale(HADS)was used for assessment to evaluate the surgical efficacy.Results There was no significant difference in depression scores before and after surgery(P＞0.05).However,postopera-tive anxiety scores,ETDQ-7 scores,and VAS scores were significantly lower than preoperative scores(P＜0.05).Among the 27 patients,9 showed significant symptom relief,13 exhibited partial relief,and 5 had no significant change compared to preoperative symptoms.The overall response rate of the treatment(significant relief and partial relief)was 81.48％(22/27).All surgeries were successfully performed.Except for secretory otitis media occurring in 2 cases,no major complications were observed.Conclusion MEACETT demonstrates significant symptom relief in PET patients,with high surgical safety and low complication rates,making it worthy of clinical promotion.

Objective To evaluate the diagnostic efficacy of the R value in tubomanometry(TMM)for the di-agnosis of patulous eustachian tube(PET).Methods The clinical data of 58 patients with PET and 65 controls were retrospectively analyzed.TMM was performed on both groups under nasopharyngeal pressures of 30,40,and 50 mbar respectively.The diagnostic efficacy of the R value for PET was evaluated through receiver operating char-acteristic(ROC)curves.Results In the control group,the average R values under nasopharyngeal pressures of 30,40,and 50 mbar were 0.86±0.50,0.76±0.41,and 0.68±0.34 respectively.In contrast,the corresponding R values in the PET group were significantly lower,which were 0.56±0.38,0.50±0.36,and 0.46±0.38 respec-tively.According to the ROC curve analysis,the areas under the curve(AUC)at these pressures were 0.62,0.74,and 0.74 respectively.The specificity and sensitivity of the R value under nasopharyngeal pressures of 30,40,and 50 mbar were 76.90％and 54.30％,74.60％and 68.10％,86.90％and 54.30％,respectively.Under pressures of 30,40,and 50 mbar,the incidence rates of R＞1 in the control group and the PET group were 29.23％(38/130)and 12.77％(12/94)(x2=8.69,P=0.003),20.00％(26/130)and 6.38％(6/94)(x2=7.20,P=0.007),10.00％(13/130)and 3.19％(3/94)(x2=2.87,P=0.09)respectively.Conclusion Although the low R value in TMM reflects the presence of PET to some extent,it does not provide adequate sensitivity and specificity to serve as an independent diagnostic criterion for PET.