Objective: To identify the mutation points of Cu/Zn superoxide dismutase(SOD1) gene in an amyotrophic lateral sclerosis(ALS) family with a unique phenotype,and to compare the value of single strand conformation polymorphism(SSCP) and denaturing high performance liquid chromatography(DHPLC). Methods: Five exons of SOD1 gene were amplified by PCR. The difference of these products were analyzed by PCR-SSCP and DHPLC.DNA sequencing was used to examine the mutation. Results: ①Mutations were found in exons 2 and 5 in several family members.DNA sequencing revealed that a base pair insertion occurred in the codon area of exon 2 and in the non-codon area of exon 5.②The results of DHPLC tests proved double peaks in one member with ALS symptoms(Ⅲ1),which indicated the possibility of mutation in SOD1 exon 4.DNA sequencing revealed that there was a heterozygote,with a mutation of GAA to GGA in exon 4 in the member with double peak. Conclusion: ①The mutations in exons 2,4,5 were proved.Insertion of exon 2 may be responsible for the disease of the ALS family in Chongqing.②Compared with PCR-SSCP,DHPLC technique has been proven to be a rapid and reliable method for screening mutation site in large samples.

Objective To identify the point mutation of Cu/Zn superoxide dismutase(SOD1) gene in an amyotrophic lateral sclerosis(ALS) family and observe the value of denaturing high performance liquid chromatography(DHPLC). Methods DHPLC and DNA sequencing were used to examine SOD1 gene of the ALS family which had not been found mutation by PCR-SSCP. Results DHPLC tests proved double peaks in one member(Ⅲ_1), Which indicated the possibility of mutation in SOD1 exon 4. DNA sequencing revealed that there was a heterozygote,with mutation of GAA to GGA in exon 4, and with a substitution of glutacid by glycine. Conclusion As compared with PCR-SSCP, DHPLC technique has proved to be a rapid and reliable method for screening mutation site in large samples.

Senna contains anthraquinones, flavonoids, polysaccharide and volatile oil and other chemical substances, which show the effect of diarrhea, antibacterial, hemostatic, regulating immune function and antioxidation. This article reviewed the Senna chemical composition, pharmacological effects, clinical application and new formulations of drug development, in order to provide reference for the research and clinical application of Senna.

Objective:To observe the clinical manifestations of a Wagner syndrome (WS) family.Methods:A retrospective clinical study. Four patients (the proband, his father, sister, and brother) and one family member (the proband's mother) from a WS family diagnosed by clinical examination in Chengdu Aidi Eye Hospital in June 2023 were included in the study. The proband's medical history was examined in detail, followed by best corrected visual acuity (BCVA), fundus color photography, optical coherence tomography (OCT), and OCT angiography (OCTA). The proband underwent full field electroretinogram (ERG) examination. The proband and his sister and brother underwent blood glucose, blood pressure, hearing, face, joint, exercise and general physical examination at the same time. Peripheral venous blood was collected from the proband and 4 other family members. The proband extracts genomic DNA samples, conducts target region capture, library construction and high-throughput sequencing after qualified quality control. The suspected pathogenic mutation sites were verified by Sanger. According to the selected mutation sites, other family members in this family were co-isolated and verified. The pathogenicity of the mutation site was analyzed using the guidelines of the American College of Medical Genetics and Genomics (ACMG).Results:Proband (Ⅱ-1) was 23 years old female. Both eyes BCVA were 0.1. The waveforms of ERG in both eyes were basically normal, and some amplitudes were reduced. Sister of the proband (Ⅱ-2) was 20 years old. Both eyes BCVA 1.0. Fundus examination showed no obvious abnormality. Brother of the proband (Ⅱ-3) was 19 years old. The left eye underwent pars plana vitrectomy combined with silicone oil filling 2 years ago due to retinal detachment and severe vitreous hyperplasia. BCVA light sensitivity, complicated cataract, and fundus opacity were observed. Right eye BCVA was 0.1. The lenses of the proband and his younger sister and brother were pointed and wedged, and the younger brother was heavier. Vitreous cavity of lens. The retina color of both eyes and the right eye of the younger brother of the protor was dark, with flaky dark areas on the side of the nose and the posterior pole, and the symmetrical retinal veil membrane hyperplasia and pulling on the periphery, showing small retinal splits. The choroidal retina showed focal and segmental symmetrically large atrophy. The optic disc was tilted. By OCT examination, the ellipsoid band was partially missing and broken, and the thickness of the choroid layer was reduced. Retinal cortical atrophy in 1 eye (younger brother of proband). By OCTA examination, the mesovascular layer of choroid was atrophied seriously and the blood density decreased. The results of laboratory and general examination of the three siblings showed no obvious abnormalities. The results of genetic testing showed that the proband, his father (Ⅱ-1), his sister and his brother carried a heterozygous mutation of the VCAN gene c.9264A>G (p.Pro3088=). According to ACMG guidelines, the pathogenicity of this variant was unknown. The mother of proband (Ⅰ-2) was wild type.Conclusions:The abnormal manifestations of WS eyes are diverse, and both anterior and posterior segments could be involved. The pathogenicity of the heterozygous variation of VCAN gene c.9264A>G (P.RO3088 =) in this family is unknown.

Objective:To analyze the clinical efficacy of scleral buckling surgery for rhegmatogenous retinal detachment (RRD) of 376 patients.Methods:A retrospective analysis was performed about 376 patients (391 eyes) who underwent scleral buckling surgery in Chengdu Aidi Eye Hospital from January 2018 to December 2019. There were 214 males (224 eyes) and 162 females (167 eyes). There were 15 binocular cases and 361 monocular cases. The average age was 37.16±16.36 years. The average course of disease was 3 months. There were 1 to more than 10 retina holes for all patients. Retinal breaks occur in all quadrants and at ora serrata. The preoperative average BCVA was 0.27 and the postoperative average BCVA was 0.41. Retinal detachment ranges was observed in 268 eyes in 1 quadrant, 97 eyes in 2-3 quadrants, 26 eyes in total, and 231 eyes with macular involvement. There were 376 eyes treated with scleral buckling, 9 eyes treated with scleral buckling combined with scleral encircling, 6 eyes treated with scleral encircling. The average follow-up time was 5 months. Postoperative follow-up was conducted to observe retinal reduction, BCVA, complications and patient compliance.Results:After the first operation, retinal reattachmnents were successfully achieved in 375 eyes (95.91%); 16 eyes (4.09%) failed in retinal reattachmnents. Eight eyes were treated with scleral buckling again, 5 eyes were treated with vitrectomy silicone oil filling, and 3 eyes were treated with air injection. After the second operation, retinal reattachmnents were ultimately achieved in 16 eyes (100.00%). The average BCVA after operation was 0.15. Postoperative intraocular pressure increased by 45 eyes (11.51%). The intraocular pressure increased from the next day to 3 days after operation. The intraocular pressure was completely controlled 1-3 days after the treatment of topical medication and 20% mannitol. Vitreous and subretinal hemorrhage in 1 eye caused by drainage of the subscleral liquid. There was no cases with postoperative infection.Conclusion:The retinal reattachment rate is 95.91% in 376 patients with RRD treated by scleral buckling surgery, and the visual acuity has significantly improved.

Objective: To investigate the clinical features and pathogenic genes of a family with osteosclerosis. Methods: Six patients and six family members from a family in Jiangsu were tested for biochemical parameters, bone metabolic markers, bone mineral density, thoracolumbar anterior lateral slices, skull positive lateral radiographs, and pelvic plain films. Meanwhile, Sanger sequencing was performed to detect gene mutations of the proband and five other family members with high bone mass. The conformation of the mutational low-density lipoprotein receptor-related protein 5 (LRP5) protein was predicted by SWISS-MODEL. Results: Four adult patients (one male and three females) were tall, with mandibular enlargement and kyphosis in the center of the lower jaw, and none of the four had fractures. Their X ray examination revealed that the skull and long bone cortex was thickened, while the sella and mandible was enlarged. In addition, the absolute values of bone mineral density at each site of all patients were significantly higher as compared with the standard age- and sex-matched adults or adolescent mean reference values, with Z scores of L2-4, femoral neck and total hip being (6.31±4.03) SD, (6.56±2.36) SD, and (7.19±2.03) SD, respectively. The results of genetic sequencing revealed that all six patients carried a heterozygous mutation (c.331G>T; D111Y) in exon 2 of LRP5 gene, while other family members showed wild type (c.331G>G; D111D). Functional prediction indicated that this mutation was located at the amino acid terminal of exon 2 of LRP5 gene, which encodes the first β-helix-generating region of LRP5 protein. Conclusion The D111Y mutation in LRP5 gene leads to a clinical phenotype characterized by benign increased bone mineral density without increasing the risk of fracture. This mutation may further affect the downstream Wnt signaling pathway by altering the spatial structure of LRP5 protein, thereby promoting maturation and differentiation of osteoblasts and resulting in osteosclerosis.

Objective: To explore the role of TLR 9 in intrauterine transmission of hepatitis B virus (HBV) through blood pathway and placenta. Methods: Epidemiological investigation was carried out in 290 HBsAg positive parturients and 45 normal parturients (control group) in Northwest Women and Children Hospital of Shaanxi Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect five serological makers of hepatitis B and TLR 9 levels in peripheral blood of pregnant women and newborns. HBV DNA was detected by real-time fluorescence quantitative PCR. Detection of TLR 9 expression in placenta by immunohistochemical method. A case-control study was conducted to analyze the difference of TLR 9 levels in placenta and peripheral blood of HBsAg- positive pregnant women with intrauterine transmission of HBV. Results: The incidence of dominant HBV infection (DBI), occult HBV infection (OBI) and intrauterine transmission of HBV were 9.28% (27/291), 40.21% (117/291) and 49.48% (144/291) respectively. (1) The level of TLR 9 in peripheral blood of HBsAg-positive parturients, non-HBV intrauterine transmission (NBIT) group and OBI group were significantly lower than that of control group (P<0.001). The level of TLR 9 in DBI group was significantly higher than those in NBIT group and OBI group (P=0.000). (2) The TLR 9 level in HBeAg-negative group was significantly lower than that in HBeAg-positive parturients in OBI group (P=0.01). (3) With the increased severity of intrauterine transmission of HBV in each HBV DNA load group, the TLR 9 level in maternal peripheral blood increased significantly (P<0.05). (4) With the increased severity of intrauterine transmission of HBV, the levels of TLR 9 increased significantly in antiviral therapy, immunoglobulin injection and non-hepatitis B vaccine groups (P<0.05). (5) The expression of TLR 9 in placenta tissues with DBI group was significantly higher than that in OBI group and NBIT group (P<0.05). Conclusions HBV can inhibit the secretion of TLR 9 in parturient to some extent, but HBeAg can stimulate the secretion of TLR 9. However, with the increased severity of intrauterine transmission of HBV, the level of TLR 9 in parturients is increased by intra-group cross-differentiation. Therefore, TLR 9 is not an independent marker for screening and grouping, but it can be used as an reference indicator for the monitoring and management of HBsAg-positive parturients.

Objective: To investigate the expression of IL-18 in peripheral blood of HBsAg positive parturients in intrauterine transmission of HBV. Methods: A case-control study was conducted in 282 HBsAg positive parturients and 43 health parturients (control group) in Northwest Women and Children Hospital of Shaanxi Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect five serological makers of hepatitis B, real time PCR was used to detect HBV DNA, and flow liquid chip method was used to detect IL-18 levels in peripheral blood of parturients and newborns. Results: The incidence of dominant HBV infection (DBI), occult HBV infection (OBI) and intrauterine transmission of HBV were 8.42% (24/285), 40.00% (114/285) and 48.42% (138/285), respectively. The level of IL-18 in peripheral blood of HBsAg-negative parturients were significantly lower than those of HBsAg-positive parturients (P=0.001), non-HBV intrauterine transmission (NBIT) group (P=0.001) and OBI group (P<0.001). The level of IL-18 in HBeAg negative group was significantly lower than that in HBeAg positive group (P=0.023). When HBV DNA load was ≥10³ copies/ml, the level of IL-18 was significantly higher than that in HBsAg-negative group (P<0.01). With the increase of HBV DNA load in maternal blood, the level of IL-18 increased (P=0.024). When HBV DNA load was 10³-10⁶ copies/ml, the level of IL-18 in DBI group was significantly lower than that in NBIT group (P=0.022), and increased with the increase of HBV DNA load in maternal blood (P=0.016). With the increased severity of intrauterine transmission of HBV, the level of IL-18 in non-hepatitis B vaccine group decreased significantly (P=0.044). The level of IL-18 in non-hepatitis B vaccine group and immunoglobulin injection group was significantly higher than that in NBIT group (P<0.05). Multivariate analysis showed that the linear relationship between maternal HBeAg status and maternal IL-18 levels had statistical significance (P=0.01). Conclusions IL-18 is a higher level balance regulator of Th1/Th2 immune network. Monitoring the level of IL-18 in HBsAg-positive parturients can be used not only for predicting the probability of DBI and OBI, but also as an intervention mean, especially for those who are HBeAg-positive and had HBV DNA load ≥10³ copies/ml, to improve maternal cellular immune function, which is conducive to interrupting intrauterine transmission and providing a theoretical basis for the prevention and control of HBV intrauterine transmission.

Objective: To investigate the influence of IFN-γ and IL-12 levels in prenatal peripheral blood of HBsAg-positive parturients on intrauterine transmission of hepatitis B virus (HBV). Methods: A case-control study was conducted in 282 HBsAg positive parturients and 43 health parturients (control group) in Northwest Women and Children Hospital of Shaanxi Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect five serological makers of hepatitis B in peripheral blood of parturients. HBV DNA was detected by real-time fluorescence quantitative PCR. Detection of cytokines IFN-γ and IL-12 levels were conducted with liquid chip-based flow cytometry method. The serum levels of five serological markers of hepatitis B and HBV DNA in 285 newborns were detected within 24 hours after birth. Results: The incidence of intrauterine dominant infection (DBI), occult infection (OBI) and intrauterine transmission of HBV in HBsAg positive parturients were 7.37% (21/285), 40.70% (116/285) and 48.07% (137/285), respectively. The level of IFN-γ in peripheral blood of HBsAg-negative parturients was significantly lower than those of HBsAg-positive parturients (t=-2.55, P=0.011), NBIT group (t=-2.54, P=0.012) and OBI group (t=-2.33, P=0.021). In HBV DNA load of 10³-10⁶ copies/ml group, the levels of IFN-γ in the DBI group were significantly lower than those in OBI group and NBIT group (P<0.01). The level of IFN-γ in maternal peripheral blood decreased significantly with the increased severity of intrauterine transmission of HBV (χ²=6.40, P=0.041). In the antiviral treatment group, the level of IL-12 in maternal peripheral blood decreased significantly with the increased severity of intrauterine transmission of HBV (χ²=8.90, P=0.012). Multivariate analysis showed that there was a significant linear relationship between maternal IFN-γ level and maternal age, placenta previa and hepatitis B vaccine injection (P<0.05). The linear relationship between the level of maternal IL-12 and the mode of rupture and hepatitis B vaccine injection had statistical significance (P<0.05). Conclusions HBV can stimulate the expression of IFN-γ and inhibit the secretion of IL-12 in pregnant and lying-in women, but the expression of IFN-γ in HBsAg-positive parturients showed intra-group differentiation, and the maternal level of IFN-γ will decrease in HBeAg-positive and high-HBV DNA loadstatus. Increasing the levels of IFN-γ and IL-12 in HBsAg-positive parturients is beneficial to block intrauterine transmission of HBV, especially DBI.