OBJECTIVE: To explore the genetic basis for a patient with Alport syndrome (AS) and confirm the existence of a splicing variant. METHODS: An AS patient diagnosed at the Affiliated Hospital of Inner Mongolia Medical University on January 8, 2021 for significant proteinuria and occult hematuria was selected as the study subject. Clinical data was collected. Peripheral blood samples were collected for the extraction of genomic DNA. Whole exome sequencing and Sanger sequencing were carried out to identify potential genetic variants. An in vitro experiment was also conducted to verify the abnormal mRNA splicing. Bioinformatic software was used to analyze the conservation of amino acids of the variant sites and simulate the 3D structure of the variant collagen IV protein. Immunofluorescence and immunohistochemistry were carried out on renal tissues from the patient to confirm the presence of AS kidney injury. RESULTS: The patient, a 21-year-old male, had a 24-hour urine protein of 3.53 g/24 h, which fulfilled the diagnostic criteria for proteinuria. His blood uric acid has also increased to 491 μmol/L. DNA sequencing revealed that he has harbored a c.835-9T>A splice variant of the COL4A5 gene, which was not found in either of his parents. In vitro experiment confirmed that the variant has removed 57 bp from the exon 15 of the mRNA of the COL4A5 gene. The deletion may cause loss of amino acid residues from positions 279 to 297, which in turn may affect the stability of the secondary structure of the α5 chain encoded by the COL4A5 gene. The amino acids are conserved across various species. The result of homology modeling indicated that the trimerization of Col-IV with the mutated α5 chain could be achieved, however, the 3D structure was severely distorted. The AS kidney damage was confirmed through immunofluorescence assays. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.835-9T>A variant was classified as likely pathogenic (PVS1_Moderate+PS3_Moderate+PM2_Supporting+PS2+PP3+PP4). CONCLUSION The c.835-9T>A variant of the COL4A5 gene probably underlay the AS in this patient. In vitro experiment has confirmed the abnormal splicing caused by the variant. Histopathological examination of the kidney tissue has provided in vivo evidence for its pathogenicity. Above finding has expanded the mutational spectrum of the COL4A5 gene.
Humans ; Male ; Young Adult ; Amino Acids ; China ; Collagen Type IV/genetics* ; Exons ; Nephritis, Hereditary/genetics* ; RNA Splicing

OBJECTIVE: To explore the genetic etiology of a fetus with Cornelia de Lange syndrome type 1. METHODS: Clinical data of the fetus was collected. Genomic DNA was extracted from amniotic fluid and peripheral blood samples of the parents and subjected to low-depth copy number variant sequencing, whole exome sequencing (WES) and Sanger sequencing. Pathogenicity of the candidate variant was predicted based on the guidelines of American College of Medical Genetics and Genomics (ACMG). Minigene assay was used to assess the effect of the variant on mRNA splicing. RESULTS: WES revealed that the fetus has harbored a heterozygous c.5808+5gG>A variant in the intron of the NIPBL gene, which was predicted to affect the mRNA splicing. The same variant was not detected in either parent. The variant was not recorded in ExAC, 1000G and dbSNP databases. Comprehensive analysis showed that the variant was deleterious and may result in skipping of exon 31 during mRNA splicing. CONCLUSION The fetus was diagnosed with Cornelia de Lange syndrome type 1. Splicing variant identified by WES may be verified by minigene assay in vitro, which can provide more evidence for the prediction of its pathogenicity.
Cell Cycle Proteins/genetics* ; De Lange Syndrome/genetics* ; Female ; Fetus ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; RNA, Messenger

Objective To evaluate the quality of the clinical trial literatures published on the treatment of COVID-19 with TCM, find out the shortcomings and put forward corresponding suggestions, in order to promote TCM against COVID-19. Methods "COVID-19", "New Coronavirus Pneumonia", "TCM", "Chinese Medicine", "Clinical Trial", "Lianhua Qingwen", "Huoxiang Zhengqi", "Jinhua Qinggan", and other keywords were used to search relevant literatures in CNKI and PubMed database. Among the all the screened relevant literatures on the treatment of COVID-19 with TCM, the literature quality was assessed according to evaluation criteria of clinical trial literatures. Results A total of 463 papers related to the treatment of COVID-19 with TCM were obtained. 440 papers on theoretical research on the network pharmacology mechanism of Chinese medicine treatment of COVID-19 were excluded. Among the 23 articles included in the quality analysis, there are 3 randomized controlled studies, 1 multi-center prospective randomized controlled trial, 2 disease case report trials, and 5 uncontrolled single arm studies, 5 controlled trials and 7 retrospective studies. In the period of more than half a year, although many clinical trial documents of TCM for the treatment of COVID-19 have been published, the clinical trial design reflected in most TCM clinical trials were not standardized. There are problems in "randomness and rationality", such as no control group, no randomization design, only case studies, no blinding method in controlling bias, and insufficient objectivity in the evaluation criteria of efficacy. All of these need to be improved. Conclusion The treatment of new coronavirus pneumonia with TCM still requires more and standardized clinical trial verifications and publications to generate strong evidence-based results, such as adding control groups, increasing sample size, and using blinding methods to increase the credibility of clinical trials.

Objective To discuss the value of oral contrast ultrasonography (OCUS) in the screening of gastric and duodenal disease in asymptomatic people in rural communities.Methods An OCUS screening was conducted in 3240 residents of Chongming County in Shanghai. After taking oral ultrasonic contrast agent, real-time observation of cardia, gastric fundus, gastric body and gastric angle, gastric antrum and duodenal bulb was conducted for each subject to detect the cardiac diameter, the body of the stomach wall thickness, pyloric canal diameter and each part of the form and structure. All subjects with abnormal sonogram were examined by gastroscopy.Kappa test was used to analyze the consistency between OCUS screening results and pathological results.Results OCUS screening detected a total of 51 cases of various types of gastric and duodenal lesions, the total detection rate was 1.57％. The coincidence rate between screening results and gastroscopy/pathology examination was 93.75％ (45/48) with a good consistency (Kappa=0.905). Five cases of gastric cancer with the detection rate of 0.15％ (5/3240) were found.Conclusion OCUS can be used for the screening of gastric and duodenal diseases in the rural community, and it is worth further research and evaluation.

BACKGROUND:Co-culture withembryonic stem cels or embryonic tissues can induce differentiation of carcinoma cels into normal epithelial cels or decreasemalignancyof carcinoma cels.Acelular embryoid bodies retain the structure and important cytokines of embryonic tissues.
OBJECTIVE:To prepare acelular embryoid bodies from mouse embryonic stem cels and to investigate their effects on differentiation of mouse Lewis lung carcinoma cels at three-dimensional culturein vitro.
METHODS:Mouse embryonic stem cels(D3)were dynamicaly cultured for 7 days to produce embryoid bodiesfolowedbydecelularization with 0.1% sodium dodecyl sulfate. Mouse Lewis lung carcinoma cels were co-cultured with acelular embryoid bodiesas test group or culturedinthree-dimensionalmatrigel mediumfor 7 days as control group, respectively. Cel proliferation and expression of E-cadherin were detected by immunohistochemical staining and western blot assay, respectively. In addition, mRNA expressions ofSlug and E-cadherin were observed using RT-PCR technology.
RESULTSAND CONCLUSION:Uniform mouse embryoid bodieswere successfuly prepared, andwere completely decelularized with sodium dodecyl sulfate. After 7-day three-dimensionalmatrigelculture, in the control group,multicelular tumor spheroidswere formed,accompanied byahigherKi67positive rate;Lewis lung carcinoma cels in the test group were repopulated in the acelular embryoid bodies showing significantly lowerKi67positive rate. Compared with the control group, the absorbance ofPaxilin in the test group was significantly smaler, and the absorbance of E-cadherin was significantly higher (P< 0.05). Besides, mRNA expressions of Slug and E-cadherin were significantly decreased and increasedin the test group compared with the control group, respectively(P< 0.05). These findings indicate that the acelular embryoid bodies can promote differentiation of mouse Lewis lung carcinoma celsinthree-dimensional culturein vitro.

Objective:Through the discussion on the purchasing mode of hospital equipment in China, fully understand the advantages and disadvantages of various procurement ways, choose reasonable acquisition method, optimize acquisition method, standardize the procurement management objective.Methods: By using the method of analogy, comparison of the four main acquisition methods. By the comparative analysis, the scope of each purchase way was confirmed. Results: Public bidding was better for more than 500000 yuan or the treasury payment items, competitive negotiation approach applies to 50000 yuan-500000 yuan purchase project. Consultation approach can be applied to 50000 yuan purchase project. Information project procurement preferred to methods of government procurement.Conclusion: Reasonable choices of purchasing and proper optimization has significant meaning for equipment purchasing and management in hospital.

Objective To evaluate the influence of hydrosalpinx on endometrial receptivity by transvaginal color doppler sonography.Methods A total of 70 patients with hydrosalpinx were randomly divided into unilateral hydrosalpinx group (observation group A)and bilateral hydros-alpinx group (observation group B),35 cases for each.Another 35 diagnosed healthy parous women served as control group.Resistance indexes (RI)of uterine artery and blood distribution types in and under endometrium among 3 groups were compared.Results Observation groups A and B were obviously higher than control group in RI on hydrosalpinx side (P <0.01),whereas observation group A was evidently higher in types A and C blood distribution and observation group B in type A blood distribution than control group (P <0.05 or P <0.01).Conclusion Hydrosalpinx may has certain impact on hemodynamics of uterine artery and increase RI,leading to decreased blood distri-bution in and under endometrium and endometrial receptivity,which can affect pregnant rate subse-quently.

Objective To evaluate the influence of hydrosalpinx on endometrial receptivity by transvaginal color doppler sonography.Methods A total of 70 patients with hydrosalpinx were randomly divided into unilateral hydrosalpinx group (observation group A)and bilateral hydros-alpinx group (observation group B),35 cases for each.Another 35 diagnosed healthy parous women served as control group.Resistance indexes (RI)of uterine artery and blood distribution types in and under endometrium among 3 groups were compared.Results Observation groups A and B were obviously higher than control group in RI on hydrosalpinx side (P <0.01),whereas observation group A was evidently higher in types A and C blood distribution and observation group B in type A blood distribution than control group (P <0.05 or P <0.01).Conclusion Hydrosalpinx may has certain impact on hemodynamics of uterine artery and increase RI,leading to decreased blood distri-bution in and under endometrium and endometrial receptivity,which can affect pregnant rate subse-quently.

Objective To explore the clinical vaue of multilayer spiral CT in diagnosis of small intestinal tumor.Methods 36 cases with small intestinal tumor were selected as the research objects,the multi-slice spiral CT imaging results were analyzed retrospectively.Results Sensitivity of multi-slice spiral CT was 94.4％,accuracy of 88.9％.Conclusion Spiral CT is a safe,simple,non-invasive,effective method in diagnosis of small bowel tumors,it has high detection rate.

OBJECTIVEThe decoction of Astragali Radix and Angelicae Sinensis Radix (A&A) has shown antifibrotic effects in rats with unilateral ureteral obstruction (UUO). The aim of this study was to track the effective parts of A&A for its renoprotective effects, according to the improvement of renal function and renal tubulointerstitial damage.

METHODA&A was sequentially extracted by using different solvents for three times and eleven different parts were gained. Wistar rats were randomly divided into Sham, UUO and the treatment groups with A&A or each part of A&A. After administration of A&A or its parts for 10 days, the levels of serum creatinin (Scr) and urea were measured. The morphological changes of kidneys were also semi-quantitatively analyzed by HE, Masson stained tissue sections, which including interstitial cell infiltration, tubular atrophy and interstitial fibrosis.

RESULTThe levels of Scr, urea were significantly increased, accompanied with severe renal damage in rats with UUO. As same as A&A, the part I in the first extraction and part IC in the second extraction were all shown to decrease the levels of Scr and urea and the index of renal interstitial damage. However, the following 4 parts extracted from IC in the third extraction were shown no effect on the above indexes.

CONCLUSIONThe extract part I and part IC could be considered as the predominant parts of A&A for its renoprotective effects, due to their improvement of renal damage in interstitial nephropathy.

Angelica sinensis ; chemistry ; Animals ; Astragalus Plant ; chemistry ; Chronic Disease ; therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; isolation & purification ; Humans ; Kidney Diseases ; drug therapy ; physiopathology ; Male ; Protective Agents ; administration & dosage ; analysis ; isolation & purification ; Random Allocation ; Rats ; Rats, Wistar