Objective:To explore the application of triplet-primer PCR (TP-PCR) for the genetic testing and prenatal diagnosis in patients with Myotonic dystrophy type 1 (DM1).Methods:A total of 60 individuals from 48 pedigrees undergoing genetic testing at the Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from May 2018 to October 2022 were selected as the study subjects. TP-PCR combined with capillary electrophoresis was applied to determine the number of CTG repeats of the DMPK gene, and prenatal testing was provided to four DM1 pedigrees. This study was approved by Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No. KS-2018-KY-36). Results:A total of 52 DM1 patients were detected, mostly with muscle weakness, muscular atrophy and myotonia as the initial symptoms, along with typical myotonic potentials. Some patients also had abnormalities of other systems. The number of abnormal CTG repeats of the DMPK gene was > 50, whilst the number of CTG repeats on the normal allele had ranged from 5 to 18. The number of the most common normal CAG repeats was 6 (30.77%, 16/52). Among the four DM1 pedigrees undergoing prenatal diagnosis, one fetus was healthy, whilst three fetuses were found to have abnormal CTG repeats (> 50 times) and diagnosed with DM1. Conclusion:TP-PCR can diagnose DM1 patients with speed and accuracy. However, this method cannot accurately determine the number of CTG repeats when it exceeds 50.

Objective To investigate the correlation of fibulin 5(FBLN5)and Rho GTPase activating protein 4(ARHGAP4)in gastric cancer tissues and their relationship with the prognosis of patients.Methods One hundred gastric cancer patients admitted to the Department of General Surgery of Shijiazhuang First Hospital were selected for the study,and gastric cancer tissues＞1 cm from the tumor margin and paracancerous tissues 5 cm from the tumor margin were collected.Retrospective analysis of the relationship between FBLN5 and ARHGAP4 expression profiles and clinicopathological indices of gastric cancer,as well as their effects on survival,was performed with univariate and Cox regression analyses of the patients'clinical data.Immunohistochemical staining was used to detect the expression of FBLN5 and ARHGAP4.The relationship between the expression levels of different FBLN5 and ARHGAP4 and the survival time of patients.Results The positive expression rate of ARHGAP4 was lower than that of paracancerent tissues,and the positive expression rate of FBLN5 was higher than that of paracancerent tissues(P＜0.05).ARHGAP4 expression was associated with tumor site,Lauren's staging,lymph node metastasis,TNM stage,differentiation degree,CE,CA19-9,CA125,and immune score(P＜0.05);FBLN5 expression was associated with Lauren's staging,lymph node metastasis,TNM stage,differentiation degree,CE,CA19-9,CA125,and immune score(P＜0.05);Kaplan-Meier analysis showed that the 3-year cumulative survival rate of FBLN5 positive group was,which was lower than that of negative group(P=0.044).The 3-year cumulative survival rate of ARHGAP4 positive patients was higher than that of negative patients(P=0.021).Cox results showed that positive ARHGAP4 was protective factor for survival(P＜0.05).Conclusion The prognosis of patients in the FBLN5-positive group is worse than that in the negative group,high expression of FBLN5 is a risk factor for survival,and the prognosis of patients with ARHGAP4 negative group is better than that in the positive group,which are closely related to the development and prognosis of gastric cancer patients.

Gastrointestinal viruses include acute gastroenteritis virus and enterovirus. These viruses are highly contagious and human populations are generally susceptible to them, and the viruses require only tens to hundreds of virus particles to cause infection. Digital polymerase chain reaction (dPCR) has the advantages of high sensitivity, strong anti-interference and direct quantification. It has shown its uniqueness in the detection of gastrointestinal viruses, especially for samples with low viral loads, which is a beneficial supplement to the real-time PCR technology. This article reviews and looks forward to the application of digital PCR technology in gastrointestinal virus detection.

In order to investigate the effects of neuroprotective peptide SNP-9 which is derived from silk fibroin hydrolysate on the injury of the blood-brain barrier in Alzheimer′s disease (AD), Aβ25-35 was used to damage brain microvascular endothelial cells bEnd.3 to establish AD injury model and drug intervention was performed.MTT assay was used to detect the effects of SNP-9 and Aβ25-35 on cell viability.RT-qPCR was used to determine the effects of SNP-9 and Aβ25-35 on the mRNA levels of tight junctions (TJs)-related ZO-1, occludin and claudin-5.Western blot was used to detect the effects of SNP-9 and Aβ25-35 on the protein levels of TNF-α, phosphorylated NF-κB, NF-κB, IκBα and RAGE.The results showed that SNP-9 reduced bEnd.3 cell damage induced by Aβ25-35, and improved the abnormal mRNA levels of ZO-1, occludin and claudin-5 in model cells.It alleviated the abnormal protein levels of TNF-α, phosphorylated NF-κB, IκBα and RAGE induced by Aβ25-35. These results suggest that SNP-9 may regulate the levels of TNF-α in model cells by influencing RAGE/NF-κB pathway, and then ameliorate TJs-related abnormalities and alleviate bEnd.3 cell injury induced by Aβ25-35.

To investigate the effect of nucleoside diphosphate kinase (NDK) on the synthesis of hyaluronic acid, nucleoside diphosphate kinase gene (ndk) was overexpressed along with the hyaluronic acid-producing genes in recombinant B.subtilis. Two engineered strains named Hp8tg and Pn8tg were constructed.Uniform hyaluronic acid (HA) could be obtained from both engineered strains.HA produced by both recombinant strains was confirmed by monosaccharide composition analysis, Fourier transform infrared spectometry and nuclear magnetic resonance spectroscopy.Inducing conditions of HA fermentation were optimized by response surface methodology.Overexpression of ndk could increase the production and molecular weight of HA by 1.3-fold and 1.1-fold, respectively. This study revealed for the first time that overexpression of ndk could relieve the inhibition effect of uridine diphosphate (UDP) on Class II HA synthase and increase the production and molecular weight of HA, which proves to be an efficient strategy for the production of HA, and the preparation of other polysaccharides.

OBJECTIVE: To assess the value of low-depth whole-genome copy number variation sequencing (CNV-seq) for the analysis of chromosomal copy number variations among fetuses with echogenic bowel (EB). METHODS: A total of 163 fetuses were included in this study. Amniotic fluid (162 cases) or chorionic villi (1 case) were collected and subjected to CNV-seq for the analysis of CNVs. RESULTS: Thirteen (8.0%) pathogenic CNVs were detected, including 9 (5.5%) aneuploidies and 4 (2.4%) CNVs. The detection rate of the isolated EB group and combined EB group were 1.7% (1/58) and 11.4% (12/105), respectively. There was a significant difference between the two groups (P < 0.05). A Xp22.1 duplication was detected in both groups, and the fetuses were predicted as female DMD carriers and born healthy. Nine cases of aneuploidies and 2 (likely) pathogenic CNVs were identified in the combined EB group, all of them have warranted induced labor. CONCLUSION The prevalence of chromosomal aneuploidies and pathogenic CNVs in fetuses with combined EB was much higher than isolated EB, and most of them may warrant termination of pregnancy. Compared with isolated EB, more attention should be paid to combined EB, for which prenatal diagnosis and genetic counseling should be carried out in time.
Amniotic Fluid ; Aneuploidy ; Chromosome Aberrations ; DNA Copy Number Variations ; Echogenic Bowel ; Female ; Humans ; Pregnancy ; Prenatal Diagnosis ; Technology

OBJECTIVE: To carry out genetic testing and prenatal diagnosis for 29 Chinese pedigrees affected with tuberous sclerosis complex (TSC) and assess efficacy of combined next generation sequencing (NGS) and multiple ligation-dependent probe amplification (MLPA) for the diagnosis. METHODS: NGS and MLPA were used in conjunct to detect variants of TSC1 and TSC2 genes among the probands of the pedigrees. Paternity test was carried out to exclude maternal DNA contamination. Prenatal diagnosis was provided to 14 couples based on the discoveries in the probands. RESULTS: Twenty-seven variants were identified in the TSC1 and TSC2 genes among the 29 pedigrees, which yielded a detection rate of 93.1%. Respectively, 5 (18.5%) and 22 (81.5%) variants were identified in the TSC1 and TSC2 genes. Twelve variants were unreported previously. Prenatal diagnosis showed that five fetuses were affected with TSC, whilst the remaining nine were unaffected. CONCLUSION Above finding has expanded the spectrum of TSC1 and TSC2 gene variants. Combined NGS and MLPA has enabled diagnosis of TSC with efficiency and accuracy.
DNA Mutational Analysis ; Female ; Genetic Testing ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; Tuberous Sclerosis/genetics* ; Tuberous Sclerosis Complex 1 Protein/genetics* ; Tuberous Sclerosis Complex 2 Protein/genetics*

Objective:To compare the efficacy of open surgery and arthroscopic assisted surgery in treatment of knee dislocations.Methods:A retrospective case-control study was conducted to analyze the clinical data of 80 patients with knee dislocations admitted to Second Hospital of Lanzhou University from May 2013 to September 2019, including 59 males and 21 females, aged 18-66 years [(42.5±11.6)years]. Open multiple ligament reconstruction was performed in 49 patients (open group) and arthroscopic assisted multiple ligament reconstruction was performed in 31 patients (arthroscopic group). The postoperative hospitalization days, incidence of complications, time needed for recovery of knee range of motion (>0°, >90°, >120°), and time to complete weight-bearing were compared between the two groups. The Lysholm score, international knee documentation committee (IKDC) subjective knee form, Tegner activity level, score of the MOS item short-form health survey (SF-36), patient satisfaction and knee range of motion were compared between the two groups at the last follow-up.Results:All the patients were followed up for 1.2-7.4 years [(3.8±1.5)years]. There was no significant difference in postoperative hospitalization days or incidence of complications between the two groups ( P>0.05). No significant difference was found in time needed for recovery of knee range of motion (>0°, >120°) or time to complete weight-bearing ( P>0.05). The time needed for recovery of knee range of motion (>90°) was 90(60, 90)days in open group and 60(30, 90)days in arthroscopic group ( P<0.05). At the last follow-up, there was no significant difference in Lysholm score, IKDC subjective score, Tegner activity level, SF-36 score, or patient satisfaction between the two groups ( P>0.05). At the last follow-up, the knee range of motion was 120°(90°, 130°) in open group and 135°(120°, 140°) in arthroscopic group ( P<0.05). Conclusion:For treatment of knee dislocations, open surgery and arthroscopic assisted surgery have similar results in the long-term effect, while arthroscopic assisted surgery has benefits in early rehabilitation and ultimately better knee range of motion.

Objective:To analyse the effect of preoperative renal function classification on early outcomes for patients with acute type A aortic dissection(AAAD) and to estimate the risk factors of postoperative major adverse events.Methods:From January 2012 to December 2019, 226 patients with AAAD who underwent total arch replacement at our institution were retrospectively analysed, including 146 males and 80 females, aged(54.4±12.5) years old. Stages of preoperative renal function were defined as follows: Normal[estimated glomerular ltration rate(eGFR)≥90 ml·min -1·1.73 m -2, 68 cases], Mild(eGFR 60-89 ml·min -1·1.73 m -2, 73 cases); Moderate(eGFR 30-59 ml·min -1·1.73 m -2, 57 cases), Severe(eGFR<30 ml·min -1·1.73 m -2, 28 cases). The independent risk factors for postoperative death were analyzed by logistic regression analysis. The area under the receiver operating characteristic curve was used to assess the efficiency of eGFR for predicting the postoperative hemodialysis. Results:In-hospital death occurred in 24(10.6%) cases. Major complications included postoperative hemodialysis in 49(21.7%) cases, stroke in 19(8.4%) cases and tracheotomy in 15(6.6%) cases. The best cut-off value of the eGFR for predicting postoperative hemodialysis was 36.5 ml·min -1·1.73 m -2(area under the receiver operating characteristic curve was 0.793). The following variables were found to be risk factors of in-hospital mortality in multivariate logistic regression analysis: serum creatinine, eGFR<30 ml·min -1·1.73 m -2, neural malperfusion, bowel malperfusion, postoperative stroke and hemodialysis. Conclusion:Total arch replacement can be safely performed in patients with AAAD and mild renal dysfunction. Preoperative renal dysfunction is a risk factor for postoperative hemodialysis, and eGFR is useful for predicting the requirement for hemodialysis after total arch replacement. The severity of preoperative renal dysfunction could greatly influence the outcomes after total arch replacement for AAAD. More importance should be attached to the assessment of preoperative renal function during clinical risk assessment.

Background: Cardiopulmonary resuscitation (CPR) strategies in COVID-19 patients differ from those in patients suffering from cardiogenic cardiac arrest. During CPR, both healthcare and non-healthcare workers who provide resuscitation are at risk of infection. The Working Group for Expert Consensus on Prevention and Cardiopulmonary Resuscitation for Cardiac Arrest in COVID-19 has developed this Chinese Expert Consensus to guide clinical practice of CPR in COVID-19 patients. Main recommendations: 1) A medical team should be assigned to evaluate severe and critical COVID-19 for early monitoring of cardiac-arrest warning signs. 2) Psychological counseling and treatment are highly recommended, since sympathetic and vagal abnormalities induced by psychological stress from the COVID-19 pandemic can induce cardiac arrest. 3) Healthcare workers should wear personal protective equipment (PPE). 4) Mouth-to-mouth ventilation should be avoided on patients suspected of having or diagnosed with COVID-19. 5) Hands-only chest compression and mechanical chest compression are recommended. 6) Tracheal-intubation procedures should be optimized and tracheal-intubation strategies should be implemented early. 7) CPR should be provided for 20-30 min. 8) Various factors should be taken into consideration such as the interests of patients and family members, ethics, transmission risks, and laws and regulations governing infectious disease control. Changes in management: The following changes or modifications to CPR strategy in COVID-19 patients are proposed: 1) Healthcare workers should wear PPE. 2) Hands-only chest compression and mechanical chest compression can be implemented to reduce or avoid the spread of viruses by aerosols. 3) Both the benefits to patients and the risk of infection should be considered. 4) Hhealthcare workers should be fully aware of and trained in CPR strategies and procedures specifically for patients with COVID-19.