Objective:To construct the recombinant eukaryotic expression vector plasmids with mutant C-kit cDNA and to study the effect of the mutant C-kit gene on cell proliferation and cell cycle in gastrointestinal stromal tumor (GIST). Methods: Wild-type C-kit cDNA was cloned from human embryonic brain tissue by RT-PCR technique.Site-directed mutagenesis of the wild type C-kit cDNA was performed according to the C-kit mutations we cloned. Recombinant plasmids were stably transfected into human embryonic kidney cell line and the cells expressing mutant C-kit were selected by special cell culture medium containing G418.Expressions of C-kit protein of the transfectants were detected by Western blot.Cell proliferation and cell cycle of the transfectants were detected by MTT clolorimetic assay and flow cytometry,respectively.Whether HEK cell with mutated C-kit cDNA could grow autonomously in nude mice or not was also detected. pcDNA3 vector transfected and recombinant plasmids with wild-type C-kit cDNA transfected HEK cell were used as the control groups. Results: The mutant C-kit cDNA was obtained by site-directed mutagenesis of the wild type C-kit cDNA. Compared with the 2 control groups ,the growth rate and proliferative activity of the HEK cells with mutant C-kit cDNA were increased significantly.The analysis of cell cycle showed that more HEK cells with mutanted C-kit cDNA remained in proliferation phase (S+G 2-M)than the groups without mutated C-kit cDNA.HEK cells with the mutated C-kit also grew autonomously in nude mice.Conclusion: Mutation of C-kit gene can increase proliferation of human cells,causing malignant transformation of human normal cells,which may play an important role in the malignant transformation of GIST.

Blepharoptosis ; etiology ; Child, Preschool ; Humans ; Male ; Mucocutaneous Lymph Node Syndrome ; complications

OBJECTIVE: To explore the genetic basis of a Chinese pedigree affected with chronic kidney disease (CKD). METHODS: A Chinese pedigree comprised of 10 individuals from four generation who had visited the First Affiliated Hospital of Dali University from August 15, 2018 to July 5, 2021 was selected as the study subject. Clinical data of the proband were collected, and a pedigree survey was conducted. The proband was subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The proband, a 41-year-old female, has been diagnosed with chronic nephritis for more than 4 years. Routine urinary examination showed proteinuria and blood creatinine of 1 130 μmol/L. Renal biopsy has revealed hyperplastic glomerulonephritis, moderate tubulointerstitial disease and renal arteriosclerosis. Her elder sister, younger brother, younger sister and mother were all diagnosed with CKD stage 5. Except for her elder sister, all of them had deceased, whilst no abnormality was found in the remainders. Genetic testing revealed that the proband and four family members had harbored a c.467G>A missense variant of the PAX2 gene. The variant has been associated with focal segmental glomerulosclerosis and classified as likely pathogenic (PS1+PP3+PP4) based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). CONCLUSION The c.167G＞A variant of the PAX2 gene probably underlay the CKD in this Chinese pedigree.
Adult ; Female ; Humans ; Male ; East Asian People ; Genetic Testing ; Mutation ; PAX2 Transcription Factor/genetics* ; Pedigree ; Renal Insufficiency, Chronic/genetics*

To perform a systematic review of the data collected from case-control studies conducted earlier to investigate the correlation between E-selectin S128R polymorphism and ischemic stroke (IS) risk among the Chinese population. The PubMed, Web of Science, Chinese biomedical literature database (CBM), Chinese databases China National Knowledge Infrastructure (CNKI), WanfangData knowledge service platform (Wanfang Data), and information resource integration service platform (VIP) Databases were searched to retrieve case-control studies on the correlation between E-selectin gene S128R polymorphism and IS from the inception of the database till June 2019. The literature was screened, data were extracted, the risk of bias was reviewed, and the studies included were assessed independently by two reviewers. Stata ver. 12.0 software (Stata Corp LLC, College Station, TX, USA) was used to perform the meta-analysis. A total of 2907 cases from eight case-control studies involving 1478 IS patients and 1429 controls were included in this study. The R allele and RS genotype in E-selectin were found to be associated with the risk of IS as per the results of the meta-analysis (R vs. S : odds ratio [OR], 2.75; 95% confidence interval [CI], 2.15-3.51; p<0.00001; RS vs. SS : OR, 2.50; 95% CI, 1.95-3.19; p<0.00001; RR+RS vs. SS : OR, 2.85, 95% CI, 2.21-3.67; p<0.00001). The E-selectin gene S128R polymorphism is likely related to IS based on the results of a meta-analysis in the Chinese population, and the R allele and RS genotype of E-selectin may be IS risk factors.

OBJECTIVE：To study the improvem ent effects of total flavonoids from Morus alba on glycolipid metabolism ， inflammation and oxidative stress in gestational diabetes mellitus （GDM）model rats ，and to investigate the potential mechanism. METHODS：After feeding high fat diet for 8 weeks，female SD rats with FBG ＜6.67 mmol/L were caged with male SD rats . Pregnant female rats were randomly divided into control group ，GDM group ，M. alba total flavonoids low-dose ，medium-dose and high-dose groups （50，100，200 mg/kg），with 10 rats in each group. Except for control group ，other groups were given intraperitoneal injection of streptozotocin 25 mg/kg once to induce GDM model. After injection ，rats in each administration group were given corresponding drugs intragastrically ，control group and GDM group were given normal saline 10 mL/kg intragastrically ， once a day ，for consecutive 18 days. The levels of FBG were determined on the 3rd，7th and 18th day of pregnancy （G3d，G7d and G18 d）；the levels of blood lipids （TG，TC，LDL-C，HDL-C）and inflammatory factors （TNF-α，IL -6，IL-8），oxidative stress indicators（MDA，SOD，GSH，CAT）in serum were determined on G 18d. The protein and mRNA expressions of PPARγ and NF-κB， the expression of AMPK mRNA and p-AMPK protein were measured by Real-time-PCR and Western blotting. RESULTS ： Compared with control group ，the levels of FBG （G3d，G7d，G18d），TG，TC，LDL-C，TNF-α，IL-6，IL-8 and MDA ，protein and mRNA expression of NF-κB in GDM group were significantly increased，while the levels of SOD ，GSH and CAT ，the expressions of PPARγ protein and mRNA，AMPK mRNA and p-AMPK fcksjf@126.com protein were significantly decreased （P＜0.01）. Compared with GDM group ，the levels of FBG （G3 d，G7 d，G18 d），TG， huawei99@163.com TC，LDL-C in M. alba total flavonoids medium-dose and high- dose groups and the levels of TNF-α，IL-6，IL-8 and MDA ，protein and mRNA expression of NF-κB in M. alba total flavonoids groups were significantly decreased ；the levels of SOD ，GSH and CAT ，the expressions of PPARγ protein and mRNA， AMPK mRNA and p-AMPK protein were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Total flavonoids from M. alba can improve the glycolipid metablism ，inflammation and oxidative stress in GDM model rats ，the mechanism of which may be related to the activation of PPARγ pathway.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

A novel series of benzyl urea analogues based on the structural modification of sorafenib were synthesized. Their in vitro antitumor activities against MX-1, HepG2, Ketr3 and HT-29 were evaluated using the standard MTT assay. While several target compounds showed inhibitory activity against multiple cancer cell lines, compound 9 was of particular interest, demonstrating IC50 values (5.69-13.6 micromol x L(-1)) comparable to those of sorafenib. Furthermore, compounds 20 and 23 showed more potent inhibitory activity against HT-29 and MX-1 when compared to sorafenib. In particular, compound 20 bearing the N-3-pyridyl moiety not only exhibited greater inhibitory activity against HT-29 cell line (IC50 3.82 micromol x L(-1)), but also had improved solubility at pH 7.2, is worthy of further investigation as a lead to identify novel antitumor agents.

Objective To observe the influence of the short effect antihypertension drugs- nifedipine and medial effect antihypertension drugs- extended release nifedipine on the blood pressure variability (BPV) in essential hypertension(EH). Methods Twenty-five EH patients were underwent 24-hour noninvasive ambulatory blood pressure monitoring (ABPM) and observed their BPV respectively before taking drugs, after taking nifedipine and extended release nifedipine. Meantime,25 normotensive controls (NC) were observed. Results (1)BPV in EH group was higher than that in controlled group and the severer the rise of blood pressure, the more obvious the increase of BPV (P 0.05). Conclusions Nifedipine could increase BPV but extended release nifedipine did not change BPV while they decreased blood pressure. Effect of extended release nifedipine was better than nifedipine in decreasing blood pressure.

Objective To evaluate the dosimetric imnpact of the fixed position of two-degrade collimator in the treatment of breast cancer after radical mastectomy using intensity-modulated radiotherapy (IMRT) technique.Methods A total of ten patients with breast cancer were treated with radical mastectomy and radiotherapy sequaciously involving the supraclavicular region and the chest wall.Two different IMRT treatment plans were designed for each patient:0°,40° and two tangential field.There was no restriction on the position of two-degrade collimator(IMRT-1) (P ＞ 0.05).The beam angles and the parameters were as same as IMRT-1,but fixed the position of the two-degrade collimator of 0° and 40° at the inferior border of the supraclavicular (IMRT-2).The dose distribution of target volume and normal tissues,conformal index (CI),and heterogeneous index (HI) were estimated with the dose volume histogram (DVH) for the two intensity modulated modes.Results The CI were 0.79 and 0.73 (Z =-2.316,P＜0.05),and the HI of the IMRT-2 plans was not different from IMRT-1 (P ＞ 0.05).Considering the dose volunes of the ipsilateral lung in two plans,the values of V5,V10,D of IMRT-2 were significantly less than those of IMRT-1 (Z =-2.805,-2.812,-2.521,P ＜ 0.05).Meanwhile,the D of the contralateral lung,D of heart and D of the contralateral breast from the IMRT-2 were all lower than those oflMRT-1 (Z=-2.666,-2.701,-2.310,P＜0.05).There was no significant difference in the values of V20,V30 of the ipsilateral lung,V30 of heart and between IMRT-1 and IMRT-2 (P ＞0.05).Conclusions Compared with IMRT-1,IMRT-2 with fixed position of the two-degrade collimator could significantly reduce the low dose region of the lung and heart.It may be used as an effective alternative for breast cancer after radical mastectomy irradiation.