Medical Molecular Biology Research Articles

Biochemical study of ZNF76 rs10947540 and SCUBE3 rs1888822 single nucleotide polymorphisms in the Egyptian patients with systemic lupus Erythematosus

ABSTRACT Systemic lupus erythematosus (SLE) is a common autoimmune disease marked by the formation of apoptotic debris and the presence of autoantibodies that target nuclear components. At this moment, the actual cause of SLE is uncertain. Genetic variables have been well proven to have a significant role in the propensity of SLE. This study aimed to investigate the effect of (ZNF76) rs (10947540) and (SCUBE) rs (1888822) gene polymorphism in patients with systemic lupus erythematosus. A case control study has been carried out at Medical Biochemistry & Molecular biology and Rheumatology unit of Internal Medicine Departments, Faculty of Medicine, Menoufia University, Egypt, for 1-year duration between 1 June 2022 and 1 June 2023. Sixty patients were females (75%) and twenty patients were males (25%). Their ages ranged from 19 to 53 years. Their disease durations ranged from 7 months to 20 years. The findings indicated that the TC genotype of the ZNF76 rs10947540 gene increases the risk of SLE by 2.274-fold, while the dominant TC + CC increases the risk by 2.472-fold, and the C allele increases the risk by 2.115-fold. Additionally, the results showed that the TT genotype of the SCUBE3 rs1888822 gene increases the risk of SLE by 3.702-fold, the dominant GT + TT increases the risk by 2.304-fold, and the T allele increases the risk by 2.089-fold, while the GT genotype increases the risk by 1.918-fold. The study revealed significant associations between the genotypes of these polymorphisms and certain clinical parameters in SLE patients. These findings highlight the potential genetic contributions to SLE susceptibility and its clinical manifestations, providing valuable insights for future research and potential personalized approaches to the management of this complex autoimmune disease.

Development of DNA-Based Lateral Flow Assay for Detection of LDLR Gene Mutation for Familial Hypercholesterolemia.

The techniques for detecting single nucleotide polymorphisms (SNP) require lengthy and complex experimental procedures and expensive instruments that may only be available in some laboratories. Thus, a deoxyribonucleic acid (DNA)-based lateral flow assay (LFA) was developed as a point-of-care test (POCT) diagnostic tool for genotyping. In this study, single nucleotide variation (E101K) in the low-density lipoprotein receptor (LDLR) gene leading to familial hypercholesterolemia (FH) was chosen as a model. Hypercholesterolemic individuals (n = 103) were selected from the Malaysian Cohort project (UKM Medical Molecular Biology Institute) while the control samples were selected from the Biobank (UKM Medical Molecular Biology Institute). The DNA samples were isolated from whole blood. Polymerase chain reaction (PCR) amplification process was performed using bifunctional labelled primers specifically designed to correspond to the variant that differentiates wild-type and mutant DNA for visual detection on LFA. The variant was confirmed using Sanger sequencing, and the sensitivity and specificity of the LFA detection method were validated using the Agena MassARRAY® technique. Out of 103 hypercholesterolemic individuals, 5 individuals (4.8%) tested positive for E101K, LDLR mutation and the rest, including healthy control individuals, tested negative. This result was concordant with Sanger sequencing and Agena MassARRAY®. These five individuals could be classified as Definite FH, as the DNA diagnosis was confirmed. The sensitivity and specificity of the variant detection by LFA is 100% compared to results using the genotyping method using Agena MassARRAY®. The developed LFA can potentially be used in the POC setting for detecting the E101K variant in the LDLR gene. This LFA can also be used to screen family members with E101K variant in the LDLR gene and is applicable for other SNP's detection.

Development of an integrated and project-based laboratory course in upper-level biochemistry and molecular biology.

An integrated and projected-based laboratory course was described, integrating interconnected knowledge points and biochemistry and molecular biology techniques on a research project-based system. The program, which served as an essential extension of theoretical courses to practice, was conducted with a sophomore of basic medical science who had completed the course in medical biochemistry and molecular biology. This course engaged students in learning "genetic manipulation" and "recombinant DNA technology" to understand the target gene's role in disease mechanics, thus altering evaluation and treatment for clinical disease. Students could master applied and advanced techniques, such as cell culture, transfection, inducing exogenous fusion protein expression, purifying protein and its concentration assay, quantitative polymerase chain reaction, and western bot analysis. This laboratory exercise links laboratory practices with the methods of current basic research. Students need to complete the experimental design report and laboratory report, which could be advantageous for improving their ability to write lab summaries and scientific papers in the future. The reliability and validity analyses were conducted on the questionnaire, and we examined students' satisfaction with the course and their gains from the course. The student feedback was generally positive, indicating that the exercise helped consolidate theoretical knowledge, increase scientific research enthusiasm, and provide a powerful tool to be a better person and make informed decisions.

Application and evaluation of the hybrid "Problem-Based Learning" model based on "Rain Classroom" in experimental courses of medical molecular biology.

With the advancement of society, the cultivation of medical professionals equipped with solid theoretical knowledge, a strong sense of innovation, and critical thinking has become a crucial goal in the reform of medical higher education. Over recent years, the hybrid Problem-Based Learning (hPBL) model, a blend of Problem-Based Learning (PBL) and Lecture-Based Learning (LBL), has emerged as a novel approach in the medical education reform landscape of China. The application and efficacy of the hPBL model in medical experimental courses have piqued the interest of medical educators. The aim of this study was to appraise the application and effectiveness of the hPBL model in the experimental course of Medical Molecular Biology at Beihua University. Utilizing the "Rain Classroom" platform, students from the Preventive Medicine and Medical Imaging programs were allocated to either the hPBL or LBL method for their Medical Molecular Biology experimental courses. The hPBL model's impact on students' performance was evaluated across four domains: experimental theory, experimental operation, experimental report, and practical application. Questionnaires were employed to gauge students' experiences and perceptions. The results indicated that the final assessment scores of the hPBL group were significantly superior to those of the LBL group. Moreover, the hPBL model effectively amplified students' self-learning capability, practical application skills, and communication competencies. Students expressed a high degree of satisfaction with this blended learning model. The hPBL model, which amalgamates PBL and LBL, has demonstrated its effectiveness in medical education. Its implementation in the experimental course of Medical Molecular Biology at Beihua University yielded positive outcomes, enhancing students' performance and satisfaction levels. Consequently, it is recommended that the hPBL model be further promulgated in other medical experimental courses.

Application of Oncolytic Poxviruses: An Emerging Paradigm in Cancer Therapy.

Despite the significant advancement of new tools and technology in the field of medical biology and molecular biology, the challenges in the treatment of most cancer types remain constant with the problem of developing resistance toward drugs and no substantial enhancement in the overall survival rate of cancer patients. Immunotherapy has shown the most promising results in different clinical and preclinical trials in the treatment of various cancer due to its higher efficacy and minimum collateral damage in many cancer patients as compared to conventional chemotherapy and radiotherapy. An oncolytic virus is a new class of immunotherapy that can selectively replicate in tumor cells and destroy them by the process of cell lysis while exerting minimum or no effect on a normal cell. Besides this, it can also activate the host's innate immune system, which generates an anti-tumor immune response to eliminate the tumor cells. Several wild types and genetically modified viruses have been investigated to show oncolytic behavior. Vaccinia virus has been studied extensively and tested for its promising oncolytic nature on various model systems and clinical trials. Recently, several engineered vaccinia viruses have been developed that express the desired genes encoded for selective penetration in tumor cells and enhanced activation of the immune system for generating anti-tumor immunity. However, further investigation is required to prove their potential and enhance their therapeutic efficacy.

Change and challenge: An online course in Medical Biochemistry and Molecular Biology.

The development of information technology and portable devices has sparked a revolution in the field of education, facilitating access to diverse educational resources and lifelong learning. In particular, the COVID-19 pandemic has accelerated the transition from face-to-face to distance teaching, which requires online education to be provided worldwide. Biochemistry and Molecular Biology are key basic medical courses in laboratory-based science that cover complicated theories and applications. The balance between traditional and online courses, and the effectiveness of online courses, are fundamental to the teaching quality of Biochemistry and Molecular Biology. In this study, we explored the concepts, designs, and practices of a new blended online course and identified potential challenges. We hope that our experiences will provide new ideas for online teaching and promote teaching reform and the development of Medical Biochemistry and Molecular Biology education.

Alpha-amyrin as an anti-biofilm agent against methicillin-resistant and vancomycin-intermediate Staphylococcus aureus

Staphylococcus aureus has caused life-threatening infections and developed resistance against conventional antimicrobials, posing a significant threat to human health worldwide. Biofilms that surround the bacteria cells act as a protective layer, allowing cells inside the biofilm to be resistant to external stresses such as antimicrobials. Therefore, biofilms further complicate treatment available for infections caused by multi-drug resistant Staphylococcus aureus. A previous study on alpha-amyrin (AM), derived from ursane, was reported to significantly reduce the biomass and inhibit the metabolic activity of reference strain methicillin-resistant and methicillin-sensitive S. aureus (MRSA and MSSA, respectively). In this study, the antibiofilm activity of AM was extended to include clinical isolates of MSSA and MRSA, and laboratory-generated vancomycin-intermediate S. aureus (VISA) collected from University Kebangsaan Malaysia Medical Center (PPUKM) and Universiti Kebangsaan Malaysia Medical Molecular Biology Institute (UMBI). Pre-formed biofilms of biofilm-forming isolates identified from the Congo Red Agar (CRA) assay were then exposed to AM, vancomycin and oxacillin, and evaluated using the crystal violet and resazurin assays. The results showed that AM reduced the biofilm biomass of three isolates of MSSA, eight isolates of MRSA and four isolates of VISA but increased the metabolic activity in certain MSSA, MRSA and VISA isolates, indicating AM may possess biofilm reduction effects but not bactericidal effects. Based on these findings, AM could be further studied and developed as a potential therapeutic agent for chronic S. aureus infections.

The journey of F1000Research since inception: through bibliometric analysis.

Background: Bibliometric analysis is an approach adopted by researchers to understand the various analytics such as year-wise publications, their citations, most impactful authors and their contributions, identification of emerging keywords, multiple themes (niche, motor, basic, and emerging or declining) etc. F1000Research is one of the Q1 category journals that publishes articles in various domains, but a detailed journal analysis is yet to be done. Methods: This study is an effort to extract the F1000Research journey information through bibliometric analysis using VOS-viewer and Biblioshiny (R-studio) interface. The F1000Research journal started its journey in 2012; since then, 5767 articles have been published until the end of 2022. Most of the published articles are from medical science, covering Biochemistry, Genetics & Molecular Biology, Immunology & Pharmacology, Toxicology & Pharmaceutics. To understand the research journey, various analyses such as publication & citation trends, leading authors, institutions, countries, most frequent keywords, bibliographic coupling between authors, countries and documents, emerging research themes, and trending keywords were performed. Results: The United States is the biggest contributor, and COVID-19 is the most commonly occurred keyword. Conclusions: The present study may help future researchers to understand the emerging medical science domain. It will also help the editors and journal to focus more on developing or emerging areas and to understand their importance towards society. Future researchers can contribute their quality research studies, focusing on emerging themes. These authors' research can guide future researchers to develop their research area around the most impacted articles. They can collaborate with them to bring that emerging theme forward.

Seed Extract of Apium Graveolens Protects Against Cisplatin-Induced Acute Hepatotoxicity in Rats

Original Article ​ ​ Year: 2023 I Volume: 5 I Issue: 3 I Pages I 1-12 ​ ​ Seed Extract of Apium Graveolens Protects Against Cisplatin-Induced Acute Hepatotoxicity in Rats ​ Naglaa Raafat AbdRaboh 1,2*, Amina Mahdy 3, Yosra Lozon 4 ​ 1 Department of Medical Biochemistry and Molecular Biology, Faculty of Medicine, Ain Shams University, Cairo, Egypt. 2 Department of Biochemistry, Dubai Medical College, Dubai, United Arab Emirates. 3 Department of Medical Pharmacology, International School of Medicine, Istanbul Medipol University, Istanbul, Turkey. 4 Department of Clinical Pharmacy, Dubai Pharmacy College, Dubai, United Arab Emirates. ​ * Corresponding Author: ​ Naglaa Raafat AbdRaboh, MD, PhD Email address: naglaoda@hotmail.com ​ Source of funding: None Conflict of interest: None ​ Submission date: 12 September 2023 ​ Acceptance date: 26 September 2023 ​ Get permission for commercial use. ​ Key words: Cisplatin, Celery Seed Extract, Interleukins, Glutathione peroxidase, Hepatotoxicity, Prostaglandin. ​ ​​ Abstract: ​ Background: Cisplatin (CP) is extensively used in treating neoplasms; with serious side effects including hepatotoxicity. Apium graveolens (celery) has potential benefits in many disorders. Aim of the study: To investigate the possible protective role of celery seeds extract (CSE) against the damaging effects of CP on the liver. Materials and Methods: The study has included five groups of rats divided into, Group 1 as normal control, group 2 received a single dose (7mg/kg, i.p.) of CP, groups 3, and 4 pretreated daily with CSE in two different doses; group 3 (250mg/kg) and group 4 (500 mg/kg), and group 5 pretreated daily with the hepato-protective agent silymarin (100mg/kg) for 17 days followed by CP injection. Results: CP markedly affected the liver function. The glutathione peroxidase, and interleukin-6 levels were significantly decreased while the levels of heme oxygenase-1, interleukin-1 alpha were markedly elevated in the tissue homogenates of CP injected rats. The administration of CSE has ameliorated all the altered biochemical and histopathological changes of the liver; the antioxidant enzyme glutathione peroxidase, IL-6 level, and prostaglandin E2 were markedly increased while IL-1 alpha was reduced. Conclusion: CSE has a protective effect against acute hepatotoxicity induced by CP administration in rats that could be through inhibiting inflammation and increasing antioxidant activities.

Different Types and Modifications of Polymerase Chain Reaction

In this study, it is aimed to provide an up-to-date overview of different types and application areas of polymerase chain reaction (PCR) developed with technological advances. PCR has accelerated scientific studies with its use in medical research and molecular biology. PCR based methods are used in many fields such as diagnosis of infectious diseases, microorganism typing, gene expression analysis, epidemiology and taxonomy fields, oncological studies, DNA cloning, analysis of point mutations, insertion of transposon elements, polymorphism studies, population typing, phylogenetic analysis, drug level analysis, and autoantibody detection. Multiplex PCR is used for simultaneous amplification of multiple targets using multiple primer pairs, consensus PCR is used for amplification of common (conserved) gene regions of genetically related microorganisms, rep-PCR is used for amplification of repetitive fingerprint DNA sequences in microbial genomes, nested PCR is used to reduce non-specific primer binding and increase sensitivity, hot start PCR is used to reduce the presence of non-specific products and primer dimers, anchored PCR is used to enable amplification of unknown gene regions using non-specific anchor primers, ligation mediated and homopolymer PCR is used for amplification of a DNA segment with a single known primer binding site, touch-down and touch-up PCR is used to prevent mismatches by regulating annealing temperature, autosticky PCR is used for amplification of DNA fragments for gene cloning using abasic primers, methylation specific PCR is used to determine the methylation patterns of cytosine residues, inverse PCR is used for sequence analysis of unknown flanking DNA regions, asymmetric PCR is used to synthesize single-stranded DNA using primers of different concentrations, in-situ PCR is used to visualize intracellular amplification in tissue sections, RAPD is used for amplifying random DNA segments using random primers and for population analysis, immuno-PCR is used for detection of low concentration amplicons by combining ELISA and PCR methods, real-time PCR is used for quantitation and monitoring of amplification with real time fluorescent signals, digital PCR is used for absolute quantitative amplification, long-range PCR for amplification of long target DNA regions, and reverse transcription PCR is used to provide amplification by synthesizing cDNA from RNA with reverse transcriptase enzyme. PCR modifications have developed rapidly throughout history. Having knowledge about these modifications will be an eye-opener for new methods to be discovered with technological developments, will contribute to selection of appropriate methods, and will increase the sensitivity and specificity of the reactions.

The Second Asia-Oceania Human Proteome Organization (AOHUPO) Online Education Series on the Renaissance of Clinical Proteomics: Biomarkers, Imaging and Therapeutics

In 2021, the Asia-Oceania Human Proteome Organization (AOHUPO) initiated a new endeavor named the AOHUPO Online Education Series with the aim to promote scientific education and collaboration, exchange of ideas and culture among the young scientists in the AO region. Following the warm participation, the AOHUPO organized the second series in 2022, with the theme “The Renaissance of Clinical Proteomics: Biomarkers, Imaging and Therapeutics”. This time, the second AOHUPO Online Education Series was hosted by the UKM Medical Molecular Biology Institute (UMBI) affiliated to the National University of Malaysia (UKM) in Kuala Lumpur, Malaysia on three consecutive Fridays (11th, 18th and 25th of March). More than 300 participants coming from 29 countries/regions registered for this 3-days event. This event provided an amalgamation of six prominent speakers and all participants whose interests lay mainly in applying MS-based and non-MS–based proteomics for clinical investigation.

Gold Nanostars Combined with the Searched Antibody for Targeted Oral Squamous Cell Carcinoma Therapy.

Oral squamous cell carcinoma (OSCC) is the most common cancer in the oral and maxillofacial region. Due to the special physiological and anatomical position of the oral cavity, the disease often has a significant impact on the chewing, swallowing, language, and breathing functions of patients. In recent years, with the development of medical molecular biology, molecular targeted therapy has received increasing clinical attention and has gradually become a new method for the treatment of malignant tumors. In this research, gold nanostars with a high photothermal effect combined with the searched targeted antibody were used for OSCC therapy. We use the data set in the public database and construct a gene co-expression module by weighted gene co-expression network analysis (WGCNA). It was found that the turquoise module and the midnight blue module had the greatest connection to tumorigenesis. Cytoscape software was used to analyze the important modules, and the top 10 genes of each module were selected; the survival analysis of the top 10 genes was carried out by gene expression profiling interactive analysis (GEPIA), which indicated that these genes (SERPINH1, MMP11, ADAM12, FADS3, SLC36A2, C1QTNF7, SCRG1, and APOBEC2) have statistical significance as key genes that are related to the tumorigenesis of OSCC. Then, the anti-SERPINH1 antibody targeted to SERPINH1 was chosen as the inhibitor and combined with gold nanostars for photothermal assisted targeted therapy. Thus, the searched key genes can be regarded as biomarkers and therapeutic targets for further precise diagnosis.

Nitric Oxide Synthase Potentiates the Resistance of Cancer Cell Lines to Anticancer Chemotherapeutics.

Despite the advancement in the fields of medical science and molecular biology, cancer is still the leading cause of death worldwide. Chemotherapy is a choice for treatment; however, the acquisition of chemoresistance is a major impediment for cancer management. Many mechanisms have been postulated regarding the acquisition of chemo-resistance in breast cancer and the impact on cellular signalling and the induction of apoptosis in tumour cells. The mechanism of the apoptotic mutation ofp53 and bcl-2 proteins is commonly associated with increased resistance to apoptosis and, therein, to chemotherapy. The current study was aimed to investigate A172 and MDA-MB-231 cancer cells'sensitivity against chemotherapeutic drugs, including cisplatin, doxorubicin, and paclitaxel with different doses. Moreover, it estimates resistance of cancer cells by evaluating Nitric Oxide Synthase (NOS) expression and evaluate its correlation with the expression profile proteins of the apoptosis regulating Bcl-2 family. Dose-dependent sensitivity to cisplatin, doxorubicin or paclitaxel was evaluated on spheroid cultured A172 and MDA-MB-231 cells lines, was measured by time-lapse microscopy over a 72h period. Expressions of two Nitric Oxide (NO) synthases isoforms (iNOS, eNOS), anti-apoptotic (Bcl-2, phospho-Bcl-2, Mcl-1, and Bcl-xL) and proapoptotic (BID, Bim, Bok, Bad, Puma, and Bax) were evaluated by Western blot. The effect of NO modulation on antiand pro-apoptotic molecule expression was also studied using Western blot. A172 cells show more resistance to chemotherapy drugs than MDA-MB-231 cancer cells, therefore, they need higher doses for apoptosis. Resistance of gliomas might be returned to higher significant expression of endothelial eNOS expression. It was clear that there is not a significant effect of NO modulation on the expression of pro- andantiapoptotic proteins on both cell lines. The present work provides a putative mechanism for the acquisition of drug resistance in breast cancer and glioma, which might be significant for clinical outcomes.

A study of the association between long non coding RNA (ULBP1 and MICA/B) expression and preeclampsia in Egypt

BackgroundPreeclampsia is associated with excessive inflammation and increased serum monokines. Although preeclampsia pathogenesis has been connected to uterine natural killer (NK) cells, natural killer group 2, member D (NKG2D) is thought to have a role in immune response mediation. NKG2D ligands (NKG2DLs) are significantly expressed in preeclampsia, and the heterogeneity of NKG2DL expression is unclear. Aim of the studyIn current study we aimed to study the expression levels of ULBP1 and MICA/B genes in women with preeclampsia. Patients & methodsThe Medical Biochemistry and Molecular Biology Department, as well as the Gynecological Department, at Menofia University's Faculty of Medicine, conducted this research. There were 120 female in all, with 80 of them having preeclampsia (40 mild and 40 severe preeclampsia). There were 40 healthy pregnant women as control group. All were chosen from the Menofia University Hospital's, Gynecological and Obstetric Departments between May 2020 and May 2021. After all subjects have given their informed permission, full history taking, clinical examination were performed on all participants and laboratory examinations include: measurement of liver function tests (AST&ALT), renal function tests (urea, creatinine, and uric acid), complete blood count, assessment of protein in urine and calculation of gene expression of ULPB1, MICA, and MICB in placental tissue by real time q PCR method. ResultsThere was significant increase in ULPB1, MICA and MICB genes expression in cases versus control and there was significant higher level of ULPB1, MICA, and MICB in severe cases compared to mild cases. ConclusionFrom this study, it could be concluded that expression of ULPB1, MICA and MICB levels might be used as biomarkers for diagnosis and detection of severity of preeclampsia.

Oligoribonucleotide interference-PCR: principles and applications.

Polymerase chain reaction (PCR) amplification of multiple templates using common primers is used widely for molecular biological research and clinical diagnosis. However, amplifying a specific DNA sequence harboring a mutation that is present in a small number of mutant cells within a large population of normal cells (e.g., as in cancer) in a tissue is difficult using the original PCR protocol. Thus, some measures are necessary to suppress amplification of background signals. To achieve this, we developed the oligoribonucleotide (ORN) interference-PCR (ORNi-PCR) technology in which an ORN (short RNA) hybridizes with a complementary DNA sequence to inhibit PCR amplification across the specific target sequence. ORNs can be prepared inexpensively, and ORNi-PCR can be carried out easily by adding ORNs to the PCR reaction mixture. Suppressing amplification of target sequences by ORNi-PCR is useful for detecting target sequence mutations. We showed that ORNi-PCR can discriminate single-nucleotide mutations in cancer cells and indel mutations introduced by genome editing. We also showed that ORNi-PCR can identify the CpG methylation status of a target sequence within bisulfite-treated DNA, and can enrich DNA sequences of interest from a DNA mixture by suppressing amplification of unwanted sequences. Thus, ORNi-PCR has many potential applications in various fields, including medical diagnosis and molecular biology. In this review, we outline the principles of the ORNi-PCR method and its use to detect nucleotide mutations in a variety of specimens.

Two novel mutations of the γ‐secretase genes in Chinese acne inversa (hidradenitis suppurativa)

Acne inversa (AI, OMIM #142690), also called hidradenitis suppurativa (HS), is a chronic, inflammatory, recurrent and debilitating skin follicular disease that usually presents after puberty with painful, deep-seated, inflamed lesions in the apocrine gland-bearing areas of the body, most commonly the axilla, inguinal and anogenital regions. The estimated prevalence in Caucasian populations ranges from 0.1% to 1%1 but rarely reported in Chinese. About 35%–40% of AI patients have a familial history, which shows an autosomal-dominant inheritance pattern.2 In 2010, our group identified loss-of-function mutations in the NCSTN, PSENEN and PSEN1 genes in six Han Chinese families, thus first reported that haploinsufficiency of genes encoding γ-secretase components as the genetic basis for familial AI.3 To date, over 40 mutations in γ-secretase genes have been identified in familial AI patients from different populations, according to the Human Gene Mutation Database (HGMD, http://www.hgmd.cf.ac.uk/). In this letter, we reported two novel mutations in NCSTN and PSENEN in two Chinese AI families. Patient 1 is a 25-year-old male presented with recurrent inflammatory papules, painful nodules, pustules, cysts and scars on the back of neck, axillae, back, upper chest, forearms and buttocks for 6 years (Figure 1A–F). With these clinical features, a diagnosis of acne inversa was made. He had been treated with isotretinoin without significant improvement. He has no positive family history. After the written informed consent was obtained, an in-house panel-based targeted next-generation sequencing revealed a novel frame shift mutation c.1555dupA in NCSTN in patient 1 (Figure 1G). NCSTN encoding nicastrin, a glycoprotein acting as a γ-secretase complex stabilizer,4 is the most commonly mutated gene reported in familial AI. This mutation generated a premature termination codon (p.T519NfsX9), which was predicted to cause loss of function of NSCTN thereby causing the disease. In family 2, there were 1 male and 4 female patients (Figure 1H). The proband (III 4) was a 25-year-old female presented with recurrent inflammatory papules, pustules and painful nodules at the age of 11 years. Other patients in this family had similar clinical manifestations and fulfilled the diagnostic criteria of AI (Figure 1I–P). All patients had normal intellectual ability and no neuropsychiatric disorder and behaviour problems. By targeted sequencing, a heterozygous nonsense mutation, c.13C>T p.R3X, was identified in PSENEN, which was never reported before. Sanger sequencing confirmed this mutation in the proband and showed that her affected sister also carried the same mutation. PSENEN encodes presenilin enhancer protein 2 (PEN2), a member of the γ-secretase enzyme complex, in which loss-of-function mutations have been reported to cause familial AI. The nonsense mutation p.R3X was classified as “pathogenic” according to the ACMG/AMP 2015 guideline5 and should be responsible for the disease in this family. In conclusion, herein, we reported two novel mutations of γ-secretase genes in two Chinese families with AI, which expanded the mutation spectrum of γ-secretase genes underlying AI. The study was supported by the National Key Research and Development Programme of China [2016YFC0905100 to XZ], the CAMS Innovation Fund for Medical Sciences (CIFMS) [2016-I2 M-1-002 to XZ], the National Natural Science Foundation of China (NSFC) [81788101 and 81230015 to XZ] and the Natural Science Foundation of Beijing [7172167 to D-LM]. The authors wish to acknowledge the constructive comments and contributions of Wei Liu, Department of Dermatology, Peking Union Medical College Hospital, Jia-Wei Liu, Department of Dermatology, Peking Union Medical College Hospital, and Rong-Rong Wang, McKusick-Zhang Center for Genetic Medicine, State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences and Chinese Academy of Medical Sciences & Peking Union Medical College. None declared. In the study, Y.T.Q. and M.X. are the co-first authors. D.L.M. and X.Z. are the co-corresponding authors and revised the manuscript and supervised the study. Y.T.Q. and D.L.M. cared for the patients. M.X. and K.Q.L. performed the next-generation sequencing and contributed to the interpretation of the results. Y.T.Q. and M.X. wrote the initial draft. All authors participated in the design of the study, clinical assessment and genomics research. All the authors take responsibility for acquisition, analysis and interpretation of the data. All authors have read and approved the final manuscript.

Integrating basic and applied researches of SARS-CoV-2 into the medical molecular biology teaching.

Molecular biology is an important course for medical postgraduates to understand the practice of modern molecular medicine. At present, molecular biology has been widely used in the prevention, diagnosis, and treatment of clinical medicine. The COVID‐19 pandemic forced all universities suspending academic activities and online teaching all over the world, which is a kind of crisis and has become people's memory of the times. The efforts from all sectors of society, include government, research institutions, and enterprises, have gave medical students a profound impact. The integration of memory of the times into molecular biology teaching will enhance students' learning interest and understanding effect.

Value of serum tumor necrosis factor-α as an early predictor of diabetic nephropathy

Objective This study was designed to evaluate serum tumor necrosis factor-α as an early predictor of diabetic nephropathy (DN). Background Owing to the high morbidity and mortality associated with DN, it is important to establish marker used as an early prediction of DN to overcome its complication. Patients and methods This study was conducted on 70 patients selected from the Inpatient Department and Outpatient Clinics of Internal Medicine Department and Medical Biochemistry and Molecular Biology Department at Menoufia University Hospital. All patients were investigated regarding urine analysis, blood urea and serum creatinine, HBA1C, lipid profile, and urine albumin-creatinine ratio. Results The 70 patients (55 males and 15 females) with diabetes mellitus were divided into 35 patients with normoalbuminuric and 35 patients with albuminuria (30–299 mg/g Cr). Ten healthy individuals (seven males and three females) were selected as a control group. The patients fulfilled the following criteria: age greater than 18 years, initial diagnosis of diabetes at greater than 30 years of age, no signs of renal diseases, no history of cardiovascular diseases, including stroke, heart diseases, and arteriosclerosis, and no symptoms of acute inflammatory diseases. All patients and control personnel enrolled in this study gave oral and written consent to see all of their investigations. Conclusion Tumor necrosis factor-α, one of the main proinflammatory cytokines, is overexpressed in DN, and this overexpression is significantly associated with evidence of renal damage.

Correlation between psoriasis area severity index score and metabolic syndrome in psoriatic patients in Menoufia University

Objective This study was to assess the association between psoriasis and metabolic syndrome (MBS) and to determine if there is an association between psoriasis severity and MBS. Background Background: Psoriasis is a chronic inflammatory disease that affects not only the skin but also other organs. Genetic factors play an important role in individual predisposition. Lately, a positive association has been confirmed between psoriasis and MBS, in Western as well as in Middle Eastern countries. Patients and methods This case–control study was conducted at the Dermatology, Andrology and Sexually Transmitted Disease and Medical Biochemistry and Molecular Biology Departments, Faculty of Medicine, Menoufia University. It included 120 patients: 60 psoriatic patients and 60 age-matched and gender-matched healthy controls. Patients were collected from Dermatology Outpatient Clinics, Menoufia University Hospitals. Clinical, biometric, and necessary laboratory evaluations were performed. Statistical analysis was performed by using the Statistical Package for the Social Sciences (SPSS version 16.0). Results The patients with psoriasis were two times more likely to have MBS as compared with controls (38.3 vs 18.0%, P < 0.001) with an odds ratio of 2.4. All components of MBS were more prevalent in psoriasis patients than in controls. The psoriasis area severity index score was greater in patients with MBS than those without MBS (10.5 11.5 vs 7.0 8.1, P = 0.05). MBS prevalence tended to be higher in the inverse type than in others (52.2 vs 32.3%; P = 0.06) and in patients with nail pitting versus those without (45.3 vs 28.2%; P = 0.03). Conclusion This study concluded that there is an association between psoriasis and MBS.

A challenge for colorectal surgeons: pathogenesis, progression and management of the secondary tumors of the ovary

A common clinical problem encountered by colorectal surgeons is the secondary tumors of the ovary (STO), particularly in young female patients. Most STO are from the digestive tract, and the known possible metastatic mechanisms include lymphatic, hematogenous, and intraperitoneal spreading. The molecular and histopathological characteristics of STO from different sites are diverse. It is particularly important to correctly identify the origin and feature of STO, which should be clarified by combining medical history, histopathology, immunohistochemistry, molecular biology, imaging and other means. The prognosis of patients with STO is poor in general. Comprehensive therapies based on surgical resection can benefit some patients. There is no specific treatment for STO at present, but not giving up easily on these patients is the right choice that every surgeon should understand.