Objective:To investigate the clinical value of targeted sequencing panel in the detection of genetic variation in neonates in neonatal intensive care unit (NICU).Methods:All neonates (≤28 d of age) admitted in the NICU (case group) and 200 full-term healthy neonates born with no obvious phenotypic abnormalities of Huzhou Maternity and Child Health Care Hospital were enrolled in this prospective study from November 2022 to January 2023. Based on a list of preventable and treatable rare diseases as well as newly screened diseases in China, a targeted sequencing panel suitable for Chinese newborns was designed to target the pathogenic genes and mutation sites associated with 601 genes and 542 diseases. Dried blood spot specimens were prepared and analyzed by the targeted sequencing panel. Pathogenic sites detected by the panel sequencing were verified using Sanger sequencing. The genetic testing results were analyzed according to the clinical features of the neonates. According to the number of primary clinical diagnosis index (including premature infants, neonatal hyperbilirubinemia, hemorrhagic diseases, neonatal infections, ventricular septal defect/patent ductus arteriosus, and others), these patients were divided into four groups with 1, 2, 3, and ≥4 diagnosis index, respectively. Chi-square test and linear correlation Chi-square test were used for statistical analysis. Results:There were 173 patients in the case group and 30.6% (53/173) of them carried pathogenic variants, including 52 positive for pathogenic genes and one with chromosome copy number variant. The positive rate of pathogenic genes was significantly higher in the case group than in the control group [30.1% (52/173) vs. 15.0% (30/200), χ 2=12.26, P<0.001]. Fourteen pathogenic genes were detected in the case group, including FLG, UGT1A1, G6PD, MYH7, AR, ABCC2, ACADS, CYP21A2, GJB2, MEFV, PAH, PKHD1, SCN4A, and HBA. In the case group, the detection rate of pathogenic variants in jaundiced neonates was higher than that in non-jaundiced neonates [35.2% (44/125) vs. 18.8% (9/48), χ 2=4.42, P=0.036]. However, there were no statistically significant differences in the detection rates of pathogenic variants between male and female infants, infants born to mothers of advanced maternal age or not, infants born to mothers with or without gestational diabetes mellitus, premature and term infants, or infants with or without hemorrhagic disorders, neonatal infections, or ventricular septal defects/patent ductus arteriosus in the case group (all P>0.05). The detection rate of pathogenic variants showed a linear increase in infants with 1, 2, 3, and ≥4 diagnosis index [21.1% (8/38), 25.4% (15/59), 38.2% (13/34), and 40.5% (17/42); linear correlation χ 2=4.84, P=0.028]. In the case group, seven genes with a high detection rate of genetic variation (including positive pathogenic genes and carriers) were UGT1A1 [had the highest detection rate, 24.9% (43/173)], GJB2, FLG, DUOX2, ABCA4, G6PD, and MUT. Seven loci with higher mutation frequency were c.211G>A(p.Gly71Arg), c.1091C>T(p.Pro364Leu), c.-41_-40dupTA, and c.686C>A(p.Pro229Gln) in the UGT1A1 gene, c.109G>A(p.Val37Ile) in the GJB2 gene, and c.12064A>T(p.Lys4022Ter) and c.3321del(p.Gly1109GlufsTer13) in the FLG gene. Conclusion:This panel sequencing can provide effective genetic testing for neonates in NICU, especially in children with complex clinical diagnosis.

Objective:To analyze the types and distribution of pathogenic variants for neonatal genetic diseases in Huzhou, Zhejiang Province.Methods:One thousand neonates (48 ~ 42 h after birth) born to Huzhou region were selected as the study subjects. Dry blood spot samples were collected from the newborns, and targeted capture high-throughput sequencing was carried out for pathogenic genes underlying 542 inherited diseases. Candidate variants were verified by Sanger sequencing.Results:Among the 1 000 newborns, the male to female ratio was 1.02 : 1.00. No pathogenic variants were detected in 253 cases, whilst 747 cases were found to carry at least one pathogenic variant, which yielded a carrier rate of 74.7%. The most frequently involved pathogenic gene was FLG, followed by GJB2, UGT1A1, USH2A and DUOX2. The variants were classified as homozygous, compound heterozygous, and hemizygous variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), 213 neonates were verified to have carried pathogenic and/or likely pathogenic variants, with a positive rate of 21.3%. The most commonly involved genes had included UGT1A1, FLG, GJB2, MEFV and G6PD. Conclusion:Newborn screening based on high-throughput sequencing technology can expand the scope of screening and improve the positive predictive value. Genetic counseling based on the results can improve the patients′ medical care and reduce neonatal mortality and childhood morbidity, while provide assistance to family members′ health management and reproductive decisions.

The popularity of prostate-specific antigen screening has led to overdiagnosis and overtreatment of prostate cancer. Active surveillance can be used as a safe and reliable treatment option for some patients with localized prostate cancer. At the same time, compared with surgery and chemoradiotherapy, active surveillance can provide these patients with a better quality of life. A variety of new monitoring methods, including PSMA PET and biomarkers, can optimize the risk stratification of prostate cancer and reduce unnecessary puncture biopsies to further improve the quality of life of patients. However, there are still great controversies about the inclusion criteria, monitoring programs, and intervention indications for active surveillance of prostate cancer, and domestic doctors and patients still lack acceptance of it. This article reviews the research progress of active surveillance in prostate cancer.

Objective To establish Cdh5-Cre ERT /Acta2-tdTomato-STOP floxed -eGFP knockin genetic tracing mice, and to investigate its application in studies on vascular endothelial cell transition in liver fibrosis. Methods Cdh5-Cre ERT mice were mated with Acta2-KI mice, and the Cdh5-Cre ERT /Acta2-KI genetic tracing mice were obtained and identified by PCR genotyping. Primary liver sinusoid endothelial cells (LSECs) were isolated and cultured, and a model of CCl 4 -induced liver fibrosis was established. LSECs and liver tissue were collected for immunofluorescent staining to observe the expression of the fluorescent proteins tdTomato and eGFP. Results After being induced by tamoxifen, LSECs and liver tissue of Cdh5-Cre ERT /Acta2-KI genetic tracing mice expressed eGFP under the conditions for epithelial-mesenchymal transdifferentiation established in vivo and in vitro, while the control group without induction expressed tdTomato alone. Conclusion The successfully established Cdh5-Cre ERT /Acta2-KI genetic tracing mice can realize the effective labeling of epithelial-mesenchymal transdifferentiation, which provides a genetic tracing basis for the diverse sources of mesenchymal myofibroblasts in liver fibrosis.

Objective To investigate the application prospect of the most extensive genome engineering pig internationally in preclinical xenotransplantation. Methods Porcine endogenous retrovirus (PERV) knockout combined with 3 major heterologous antigen gene knockouts and 9 humanized genes for inhibition of complement activation, regulation of coagulation disorders, anti-inflammatory and anti-phagocytosis were transferred into a pig (PERV-KO/3-KO/9-TG) as a donor, and the heart, liver and kidney were obtained and transplanted to 3 Rhesus macaque recipients respectively to establish a preclinical research model of pig-to-Rhesus macaque xenotransplantation. The functional status of xenografts after blood flow reconstruction was observed and the survival of recipients was summarized. The hemodynamics of xenografts were monitored. The change of hematological indexes of each recipient was compared. The histopathological manifestation of xenografts was observed. Results After the blood flow was reconstructed, all xenografts showed ruddy color, soft texture and good perfusion. The transplant heart, liver and kidney showed full arterial and venous blood flow and good perfusion at 1 d after operation. The postoperative survival time of heart, liver, and kidney transplant recipients was 7, 26, and 1 d, respectively. The levels of creatine kinase, creatine kinase isoenzyme, and lactate dehydrogenase increased in heart transplant recipient at 1 d after operation, and gradually recovered to near normal levels at 6 d after operation. All indexes increased sharply at 7 d after operation. The level of aspartate aminotransferase increased in liver transplant recipients at 2 d after operation, and the alanine aminotransferase basically returned to normal at 10 d after operation, but the total bilirubin continued to increase. Both aspartate aminotransferase and alanine aminotransferase increased at 12 d after operation, and reached a peak at 15 d after operation. The kidney transplant recipient developed mild proteinuria at 1 d after operation, and died of sudden severe arrhythmia. Histopathology showed that the tissue structure of cardiac and renal xenografts was close to normal, and liver xenografts presented with patchy necrosis, the liver tissue structure was disordered, accompanied by inflammatory damage, interstitial hemorrhage and thrombotic microangiopathy. Conclusions PERV-KO/3-KO/9-TG pig shows advantages in overcoming hyperacute rejection, mitigating humoral rejection and coagulation dysregulation. However, whether it can be used as potential donor for clinical xenotransplantation needs further evaluation.

Objective:To investigate the clinical efficacy of percutaneous endoscopic lumbar discectomy (PELD) with the aid of neurophysiological monitoring under general anesthesia.Methods:From August 2016 to October 2019, 58 patients underwent PELD under general anesthesia were selected in our hospital; 30 were via transformational approach and 28 were via interlaminar approach. The whole operative procedures were performed under continuous monitoring of spontaneous electromyography (SEMG), and the peak value, waveform and motor unit of SEMG at the surgical side were observed. The clinical outcomes were evaluated by visual analogue scale (VAS) and Oswestry disability index (ODI).Results:PELD was successfully performed in all 58 patients. Abnormal SEMG reactions were recorded in 8 patients (13.8%), manifested as clustered or frequent high amplitude action potentials; 5 patients (16.7%, 5/30) were via transformational approach and 3 (10.7%, 3/28) were via interlaminar approach. Two patients relapsed at 3 and 6 weeks after surgery, respectively; one was treated with PELD again and the other one was treated with lumbar fusion and instrument fixation. The pain at the lumbago and leg was alleviated obviously after surgery in the 56 patients; the VAS scores were 7.43±1.32, 2.55±0.87 and 1.59±0.87 before surgery, and 3 d and 3 months after surgery, respectively, with significant differences ( P<0.05); the mean ODI before surgery and 3 months after surgery were 67.36±7.13 and 12.39±5.48, respectively, with significant difference ( P <0.05). Conclusion:PELD with the aid of neurophysiological monitoring under general anesthesia is safe and reliable, which can achieve good clinical efficacy.

OBJECTIVE:To screen the α-glucosidase inhibitory active part from Pothos chinensis. METHODS:The aqueous extractions of P. chinensis were extracted by petroleum,ethyl acetate,n-butyl alcohol in turn to obtain different polarparts. Effect of each part on α-glucosidase inhibitory activity was determined,and enzyme inhibition kinetics was conducted for the screened parts with strong activity and relatively high yield rate;effects of each part on blood glucose level of mice loaded with glucose,su-crose and starch were respectively determined (using Acacoside tablet as positive control). RESULTS:Enzyme inhibition kinetics in vitro showed the ethyl acetate part [yield rate was 0.40%,enzyme activity inhibition rate was(72.90±2.85)%] had strongα-glu-cosidase inhibitory activity and showed a dose-dependent,fast,non-competitive and reversible model. Results of in vivo glucose tol-erance indicated that Acacoside tablet and each part of P. chinensis had no effects on blood glucose level of glucose-loaded mice (P>0.05);while Acacoside tablet and ethyl acetate part in P. chinensis could reduce 30,60 min blood glucose level of su-crose-loaded mice and 30,60,120 min blood glucose level of starch-loaded mice(P<0.05 or P<0.01). CONCLUSIONS:Ethyl acetate part is theα-glucosidase inhibitory active part from Yao medicine P. chinensis.

Gallbladder carcinoma is the most common malignancy in biliary tract.Early dissemination,rapid and silent progression and delayed diagnose could portend a dismal prognosis.The 5-year survival rate is less than 10％.Clarifying the etiological feature can improve the early diagnosis.Comprehending surgical indications of benign lesions can provide the methods for rational prophylactic resections.Standardized treatment of accident gallbladder carcinoma could improve the patients' prognosis.Efficient serologic markers could be used for early diagnosis and screening.

Objective To discuss the correlation between the joint detection of BA,EMAb,ACA and unexplained recurrent spontaneous abortion(URSA).Methods Selected 144 URSA cases,166 SA(sporadic abortion)cases,188 normal pregnant women,and detected their BA,EMAb,ACA and statistical analysis of parallelism.Results Comparing URSA group with SA group and control group respectively,there were significant statistical difference between BA negative rate (74.31%, 25.30% and 50.53%),EMAb positive rate (35.42%,7.23% and 6.91%)and ACA positive rate (26.39%,5.42% and 5.32%)(χ2 = 19.344 ~ 74.180;P < 0.05;χ2 = 37.837 ~ 42.586,P < 0.05),(χ2 = 26.355 ~ 29.270,P < 0.05).URSA group’s BA,ACA and EMAb joint detection rate of positive (84.72%)was significantly higher than SA group and the con-trol group (χ2 =35.532~93.076,P <0.05).Conclusion There was a close relationship between the lack of BA,the genera-tion of EMAb,ACA and unexplained recurrent spontaneous abortion.Joint detection has important clinical significance in auxiliary diagnosis of unexplained recurrent spontaneous abortion and prognosis judgement.

Objective: To explore the effects of monocrotaline (MCT) on right ventricular function and expression of cardiac canonical transient receptor potential channels (TRPC) subfamily in experimental rats.
Methods: The SD male rats were randomly divided into 2 groups:Control group, the rats were normally fed and MCT group, the rats received a single dose injection of MCT 60 mg/kg to induce myocardial hypertrophy. n=10 in each group and all animals were treated for 3 weeks. The right ventricular hemodynamics parameters and right ventricular hypertrophy index (RVHI) were measured, right ventricular myocardium tissue section was observed by HE staining, the mRNA and protein expressions of TRPC subfamily were examined by RT-PCR and Western blot analysis.
Results: Compared with Control group, MCT group had increased RVSP, RVHI, RV+dp/dtmax and decreased RV-dp/dtmax, all P<0.01. In MCT group, the right ventricular myocardial cells had the thinker ifber, deeply stained nuclei with irregular shape. Compared with Control group, the mRNA and protein expressions of right ventricular TRPC6 were elevated in MCT group, n=6 and n=4 respectively, all P<0.05.
Conclusion:Right ventricular hypertrophy could be induced by 3 weeks MCT treatment, it up-regulating the mRNA and protein expressions of TRPC6 which might be involved in the occurrence and development of cardiac hypertrophy in experimental rats.