Objective:To explore the clinical application of preimplantation genetic testing for monogenic disorders (PGT-M) in an unique case with Spinal muscular atrophy (SMA) type 2+ 0.Methods:A special SMA family presented at the Third Affiliated Hospital of Guangzhou Medical University on October 19, 2020 was selected as the study subject. Multiple ligation-dependent probe amplification (MLPA) and molecular tagging linkage analysis were carried out to identify the SMN1 genotype of the couple and their fetus. Subsequently, next-generation sequencing (NGS), molecular tagging linkage analysis, and chromosomal microarray analysis were employed to determine the haplotypes and validate the result of PGT-M on the 11 embryos derived for the couple. Results:The female partner was identified as a carrier of the rare SMN1[2+ 0] variant, and prenatal diagnosis confirmed the fetus to be affected by SMA. Ultimately, PGT-M has successfully selected four embryos free from the pathogenic SMN1 variants and X chromosome deletion. Conclusion:PGT-M can effectively prevent the transmission of rare genetic variants such as the SMA 2+ 0 subtype in the families. Above finding has provided guidance for genetic counseling and family planning for the couple.

The presence of replication-competent HBV DNA in the liver and/or serum of HBsAg-negative individuals is a sufficient and necessary condition for the diagnosis of occult hepatitis B virus infection (OBI). In recent years, Chinese scholars have proposed what is considered a more "rigorous" definition, i.e., on this basis, HBV window period (WP) infection is excluded, which corresponds to a serum HBV DNA level of below the lower limit of detection or a low positive value (< 200 IU/mL). As the definition of WP for HBV infection remains unclear and its duration is highly variable, "HBV DNA < 200 IU/mL" is not the only criterion in OBI patients. Therefore, it is believed that there is still a lack of sufficient basis and operability for the definition of OBI based on "the exclusion of HBV WP infection" and "HBV DNA < 200 IU/mL" as "rigorous" conditions for the diagnosis of OBI.

Objective To establish a PCR-based capillary electrophoresis(PCR/CE)to detect Survival Motor Neuron 1(SMN1)and Survival Motor Neuron 2(SMN2)genes and to evaluate its performance.Methods PCR/CE and Multiplex Ligation-dependent Probe Amplification(MLPA)for SMA gene diagnosis were used to blindly test the samples in sync.The performance of PCR/CE was assessed using MLPA results as the standard.Results A total of 336 samples were included in this study,consisting of 50 homozygous deletion types(14.9%),65 heterozygous deletion types(19.3%),and 221 non-deletion types(65.8%).The results of PCR/CE for detect-ing SMN1 and SMN2 copy numbers(0,1,2,3,≥4)were in complete agreement with the results of the MLPA.Conclusions PCR/CE for gene testing related to SMA could accurately detect copy numbers of exon 7 and exon 8 of the SMN1 and SMN2 genes(0,1,2,3,≥4).

Objective:Plateletcrit (PCT) is considered a new potential index to predict the degree of liver fibrosis in patients with chronic hepatitis B (CHB). This study aimed to explore the predictive value of PCT for the degree of liver fibrosis in patients with chronic hepatitis B virus (HBV) infection with alanine aminotransferase (ALT) < 2× upper limit of normal (ULN). Measurement data were compared using the t-test, ANOVA, or non-parametric test (Mann-Whitney U test). Categorical variables were compared using χ2 test or Fisher’s exact test. Methods:140 cases with chronic HBV infection who underwent liver biopsy and ALT < 2×ULN were enrolled from January 2016 to March 2021. Univariate and multivariate logistic regression and the area under the receiver operating characteristic curve (AUC) were used to determine the predictive value of PCT for the degree of liver fibrosis. The likelihood ratio (LR) was used to optimize the selection of the diagnostic cut-off.Results:(1) Among the 140 cases, there were 34 (24.3%) cases in the S0 stage, 47 (33.6%) cases in the S1 stage, 16 (11.4%) cases in the S2 stage, 19 (13.6%) cases in the S3 stage, and 24 (17.1%) cases in the S4 stage. The overall mean PCT level was 0.19 ± 0.06%. (2) Univariate analysis revealed that PCT between patients with stages of liver fibrosis was S(0-1) and S(2-4) (0.20% ± 0.05% vs. 0.16% ± 0.06%, t = 3.955, P < 0.001), S(0 -2) and S(3-4) (0.20% ± 0.05% vs. 0.15% ± 0.06%, t = 5.631, P < 0.001) and S(0-3) and S4 (0.20% ± 0.05% vs. 0.12% ± 0.05%, t = 7.113, P < 0.001), respectively, and the differences were statistically significant. Multivariate logistic regression analysis showed that PCT was an independent risk factor for liver fibrosis stages S(2-4), S(3-4), and S4 ( OR = 0.925, 95% CI: 0.859 – 0.997, P = 0.042; OR = 0.867, 95% CI: 0.789 – 0.954, P = 0.003; OR = 0.708, 95% CI: 0.593 – 0.846; P < 0.001). (3) The AUCs of PCT were 0.702, 0.777, and 0.885 for diagnosing liver fibrosis stages S(2-4), S(3-4), and S4 in patients with chronic HBV infection with ALT < 2×ULN. PCT was superior for the cirrhosis (S4) diagnosis. 92 (65.7%) cases were diagnosed as cirrhosis or non-cirrhosis according to the LR optimized diagnostic and exclusion diagnostic cut-offs (≤0.09%, ≤0.17%), with an accuracy of 97.8%. Conclusion:PCT has a high diagnostic and exclusion value for cirrhotic patients with chronic HBV infection with ALT < 2×ULN. Furthermore, it can be used as a non-invasive diagnostic index for determining and assisting the diagnosis of cirrhosis in resource-constrained areas, reducing the need for pathological examination of liver biopsies, and it has the advantage of being simple and intuitive without complex calculations.

Objective:To investigate the value of single sperm sequencing combined with PCR-reverse dot blot (PCR-RDB) technology in preimplantation genetic testing (PGT) of Southeast Asian deletion type α thalassemia.Methods:A couple of Southeast Asian deletion type α thalassemia carrier was selected in this case, who asked for assisted reproduction in the Department of Obstetrics and Gynecology of the Third Affiliated Hospital of Guangzhou Medical University on April 24, 2020. The male carrier was subjected to single sperm isolation by the swim-up method combined with micropipette due to his incomplete pedigree. Five single sperm samples were isolated and their whole-genome were amplified. The genotype of thalassemia of single sperm samples were determined by PCR-RDB. SNPs were selected as genetic markers in the range of 1 Mb upstream and downstream of -- SEA region to construct chromosomal haplotype. Next, we performed whole-genome amplification on six blastocyst biopsy samples, and then next-generation sequencing was carried out to detect the carrier status of the embryos. Finally, the non-pathogenic blastocysts were selected for transplantation, and the preimplantation genetic testing for monogenic (PGT-M) results were confirmed by the prenatal genetic diagnosis during pregnancy. Results:The pathogenic variant in the female was inherited from her mother, and four of the five single sperm samples were wild-type. Ten male informative SNP markers were identified by single sperm sequencing, and six female informative SNP markers were identified by linkage analysis of female family. PGT-M results indicated that four blastocysts were αα/-- SEA and two were -- SEA/-- SEA. The result of prenatal diagnosis revealed that the fetus was a carrier of -- SEA , which was consistent with PGT-M result, and a healthy girl was delivered at 40 weeks of gestation. Conclusion:For male carriers of Southeast Asian deletion type α thalassemia with incomplete pedigree, single sperm sequencing combined with PCR-RDB technology can be used to select SNP sites, and PGT-M can be performed by linkage analysis.

Objective:To investigate the value of single sperm sequencing combined with PCR-reverse dot blot (PCR-RDB) technology in preimplantation genetic testing (PGT) of Southeast Asian deletion type α thalassemia.Methods:A couple of Southeast Asian deletion type α thalassemia carrier was selected in this case, who asked for assisted reproduction in the Department of Obstetrics and Gynecology of the Third Affiliated Hospital of Guangzhou Medical University on April 24, 2020. The male carrier was subjected to single sperm isolation by the swim-up method combined with micropipette due to his incomplete pedigree. Five single sperm samples were isolated and their whole-genome were amplified. The genotype of thalassemia of single sperm samples were determined by PCR-RDB. SNPs were selected as genetic markers in the range of 1 Mb upstream and downstream of -- SEA region to construct chromosomal haplotype. Next, we performed whole-genome amplification on six blastocyst biopsy samples, and then next-generation sequencing was carried out to detect the carrier status of the embryos. Finally, the non-pathogenic blastocysts were selected for transplantation, and the preimplantation genetic testing for monogenic (PGT-M) results were confirmed by the prenatal genetic diagnosis during pregnancy. Results:The pathogenic variant in the female was inherited from her mother, and four of the five single sperm samples were wild-type. Ten male informative SNP markers were identified by single sperm sequencing, and six female informative SNP markers were identified by linkage analysis of female family. PGT-M results indicated that four blastocysts were αα/-- SEA and two were -- SEA/-- SEA. The result of prenatal diagnosis revealed that the fetus was a carrier of -- SEA , which was consistent with PGT-M result, and a healthy girl was delivered at 40 weeks of gestation. Conclusion:For male carriers of Southeast Asian deletion type α thalassemia with incomplete pedigree, single sperm sequencing combined with PCR-RDB technology can be used to select SNP sites, and PGT-M can be performed by linkage analysis.

Objective:To explore the necessity of lowering the cut-off value of alanine aminotransferase (ALT) in identifying chronic HBV infection patients with significant liver histological changes.Methods:The clinical data of 123 chronic HBV infection patients with normal ALT according to domestic criteria who underwent liver biopsy in the Department of Hepatology of Taizhou People’s Hospital from June 2016 to March 2021 were analyzed retrospectively. According to the cut-off ALT values recommended by 2018 version of AASLD guidelines (male 35 U/L, female 25 U/L), the patients were divided into two groups: high normal value group (HNALT, ALT≥AASLD and

With the improvement in people's awareness of diseases and the level of diagnosis and treatment, the incidence rates of immunoglobulin G4-related disease (IgG4-RD) and autoimmune liver disease (AILD) are constantly increasing, and IgG4-related sclerosing cholangitis is the overlapping part of the disease spectrum of IgG4-RD and AILD, while the association between IgG4-related autoimmune hepatitis (IgG4-AIH) and these two diseases remains unclear. This article reviews the hepatic manifestation of IgG4-RD, summarizes the clinical features of IgG4-AIH, and analyzes whether IgG4-AIH is a subtype of AIH or a hepatic involvement of IgG4-RD. It is believed that IgG4-AIH has similar but different clinical manifestations and histopathological features from classical AIH, and IgG4-AIH may be classified as two types, i.e., a subtype of AIH and the liver manifestation of IgG4-RD. The research on the pathogenesis, clinical features, and clinical diagnosis and treatment of IgG4-AIH should be taken seriously in future.

Objective:To analyze the status of hepatitis B virus (HBV) infection in pregnant women in Taizhou City in recent years and the effect of immunization management of hepatitis B vaccine project on the status.Methods:The pregnant women hospitalized in Taizhou People′s Hospital, Taizhou Second People′s Hospital, Taizhou Traditional Chinese Medicine Hospital and Taixing People′s Hospital from 2014 to 2017 were enrolled. According to the HBV serological results, the pregnant women were divided into non-immune population, successful immunization population, previous HBV infection population, HBV infection population and atypical manifestation population. The year of immunization management for the implementation of the hepatitis B vaccine plan was 1992. The HBV infection status of the pregnant women was analyzed based on the year of delivery and vaccination status, respectively. Chi-square test and trend chi-square test were used for statistical analysis.Results:A total of 31 449 cases were included in this study, of which 13 203 (41.98%) were non-immunized, 10 123 (32.19%) were successfully immunized, 6 409 (20.38%) were previous HBV infected, 1 566(4.98%) were HBV infected, and 148(0.47%) cases were atypical manifestation. The negative rate of all HBV serological markers of pregnant women born before 1992 and after 1992 (including 1992) were 42.07%(10 794/25 657) and 41.59%(2 409/5 792), respectively, with no statistically significant difference ( χ2=0.44, P=0.51). The hepatitis B surface antibody (anti-HBs) positive rate of pregnant women born before 1992 was 28.95%(7 428/25 657), which was lower than 46.53%(2 695/5 792) of pregnant women born after 1992 (including 1992). The difference was statistically significant ( χ2=668.94, P<0.01), and showed an upward trend year by year ( χ2=602.11, P<0.01). The hepatitis B core antibody (anti-HBc) positive rate of pregnant women born after 1992 (including 1992) was 8.81%(510/5 792), which was lower than 22.99%(5 899/25 657) of pregnant women born before 1992, the difference was statistically significant ( χ2=589.10, P<0.01), and the overall trend was declining year by year ( χ2=478.72, P<0.01). The hepatitis B surface antigen (HBsAg) positive rate of pregnant women born before 1992 was 5.46%(1 402/25 657), which was higher than 2.83%(164/5 792) of pregnant women born after 1992 (including 1992), the difference was statistically significant ( χ2 =69.23, P <0.01), and the overall trend was decreasing ( χ2=49.25, P<0.01). Among pregnant women infected with HBV, the negative rate of hepatitis B e antigen (HBeAg) was 78.16%(1 224/1 566). Conclusions:Pregnant women with HBV infection in Taizhou City are mainly HBeAg negative. Hepatitis B vaccine immunization management significantly reduces the HBsAg positive rate and anti-HBc positive rate of pregnant women, and increases the positive rate of anti-HBs, while the rate of all HBV serum marker negative is not significantly decreased. Horizontal transmission may still be a risk factor for HBV present and previous infections.