Objectives: To study the e ect of human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang on human osteosarcoma mice model. Methods: 40 standardized human osteosarcoma mice model were divided into 5 groups, treated with PBS, Paav-MCS, human P27KIP1 gene AVV virus, Chinese herb Pien Tze Huang or human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang in each di erent group. Results: ① Mice model of transplant human osteosarcoma were established successfully. ②Osteosarcoma were inhibited successfully by human P27KIP1 gene AVV virus, Chinese herb Pien Tze Huang or human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang. ③Mice model of transplant human osteosarcoma were active during observation. Conclusions: ①These results suggested that human P27KIP1 gene AVV virus, herb Pien Tze Huang and human P27KIP1 gene AVV virus combined with herb Pien Tze Huang were e ective on human osteosarcoma. ②Among them, the most e ective treatment was human P27KIP1 gene AVV virus combining with herb Pien Tze Huang. ③Chinese herb Pien Tze Huang was possible useful for life improvement of osteosarcoma patient.

ObjectiveThe aim was to construct the recombinant plasmid of pET-28a-G-protein pathway suppressor 2 (GPS2) GPS2,express GPS2 protein in E.coli,and obtain specific polyclonal antiserum of GPS2.MethodsGPS2 gene was obtained and the amplified fragment was then cloned into E.coli expression vector pET-28a to construct recombinant plasmid.The recombinant plasmid was transformed into E.coli expression strain BL21(DE3).IPTG induces the expression protein GPS2 protein,and the induction conditions were optimized.The induced product was purified by Ni2+ affinity chromatography,and the purified product was dialyzed with buffer for refolding.The purified protein can be used as antigen,injected to immunize male New Zealand white rabbit to get polyclonal antiserum.The titer and specificity of the rabbit antiserum were detected by ELISA and Western Blotting.ResultsThe E.coli expression vector pET-28a-GPS2 was constructed successfully and the recombinant protein was efficiently expressed and purified.The purified protein was used to immunize male New Zealand white rabbit to get polyclonal antiserum and the ELISA and Western Blot results showed that the high titer of specific polyclonal antiserum.ConclusionGSP2 could be highly expressed in E.coli.Antiserum of GPS2 protein can be obtained by the purified recombinant to analyze its function.

Objective To investigate the safety and efficacy of photoselective green laser vaporization of bladder tumor (PVBT). Methods A total of 522 patients with bladder tumor were enrolled in present study from January 2010 to May 2015, including 405 cases of non muscle-invasive bladder cancer (NMIBC) and 117 cases of muscle-invasive bladder cancer (MIBC). All of patients were treated with PVBT and intravesical instillation of epirubicin. Patients with MIBC received intravenous chemotherapy (kisi-hama and cisplatin). Results The hospitalization time was (7.32±1.28) days, the operation time was (27.08±5.36) min, and the indwelling urinary catheter was (2.42±0.34) days for patients in NMIBC group. During the follow-up period (12-60 months), 38 cases (9.4％) relapsed, of which 3 cases underwent radical cystectomy, and other 35 cases underwent PVBT again. All 405 patients were alive at the end of follow-ups. The hospitalization time was(26.18 ± 1.92) days, the operation time was (38.32 ± 6.54) min, and the time of indwelling urinary catheter was (2.72 ± 0.85) days for patients of MIBC group. During the follow-up period (12-60 months), 19 cases (16.2％) relapsed. Among them, 4 patients underwent radical cystectomy, and other 15 cases underwent PVBT. Six patients died from distant organ metastasis (including 2 cases of pulmonary metastasis and 4 cases of bone metastasis), and other 111 patients survived. Conclusion PVBT is safe and effective in the clinical application, especially for NMIBC and MIBC patients who are unable or unwilling to undergo radical cystectomy.

This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.

Objective To study the effects of fibroblast growth factor 2 (FGF2) on the proliferation and gene expression profiles of rabbit articular chondrocytes in vitro after different time periods of stimulation.Methods The chondrocytes were isolated and cultured in vitro,and the 3rd generation cells were harvested.Cells were divided into three groups.In the group 1 (FGF2 short-time action group),chondrocytes were cultured in medium with FGF2 for one day,and then transferred to fresh culture medium without FGF2 and cultured for another 6 days.In the group 2 (FGF2 long-time action group),chondrocytes were cultured in medium with FGF2 for 7 days.In the Group 3 (control group),chondrocytes were cultured in culture medium without FGF2 for 7 days.After culture for 1,3,and 7 days,the proliferation of chondrocytes in the all groups was detected respectively.Following extraction of mRNA,the gene expressions of BMP2,BMP4,SOX9 and COL2A1 of the chondrocytes in the all groups were determined by quantitative real-time polymerase chain reaction (qRT-PCR).The content of type Ⅱ collagen was measured via immunofluorescence staining.Results Compared with the control group,FGF2 promoted the proliferation of chondrocytes in the short-and long-time action groups and there was no significant difference between the two FGF2-treated groups.The results of qRT-PCR indicated that different treatment induced different gene expression profile.Particularly,compared with the control group and the FGF2 long-time action group,the expression of BMP2,BMP4,SOX9 and COL2A1 in the short-time action group were significantly upregulated at the 7th day.Immunofluorescence intensity of type Ⅱ collagen in the group 1 was stronger than that in the control group and group 2.Conclusions Different administration of FGF2 for different time periods induced different responses of chondrocytes.Short-term FGF2 stimulation was more beneficial to maintain the phenotype of chondrocytes and the synthesis of extracellular matrix.

BACKGROUND:Previous studies have shown that minichromosome maintenance 3 is related with fluorosis, but the expression of minichromosome maintenance 3 in fluorosis patients is not clear yet.
OBJECTIVE:To analyze the mRNA expression level of minichromosome maintenance 3 in peripheral blood from patients exposed to fluoride and normal controls.
METHODS:Eleven patients with mild fluorosis by drinking water (exposure group) and 11 cases of control (non-exposure group) were selected for research. SYBRGreen1 real-time quantitative PCR was used to determine the mRNA expression of minichromosome maintenance 3 in peripheral blood mononuclear cel s, and the liver and renal function indicators were detected.
RESULTS AND CONCLUSION:mRNA expressions of minichromosome maintenance 3 in the exposure group and non-exposure group were (0.573 60±0.102 59) and (0.550 0±0.171 81), respectively, and there was no significant difference between two groups (P>0.05). There were no significant differences in the liver and renal function indicators between two groups. The results indicate that mild fluorosis has no significant effect on mRNA expression of minichromosome maintenance 3 in the peripheral blood mononuclear cel s. More indicators are needed to compressively analyze the effect of fluoride on the liver and renal functions.

Objective To investigate the clinical effect of irradiated-host bone ends' union after transplantation of pedicled vascularized fibular periosteum in the treatment of Children's tibia sarcoma.Methods From June,2016 to December,2016,there were 5 children of tibia sarcoma,which were 2 boys and 3 girls,aged of 9-15 years (mean,12 years).They were treated by the re-transplantation of extracorporal irradiated segmental autograft,and used ipsilateral pedicled vascularized fibular periosteum cover the ends of irradiated-host bone to shorten the bone union time of irradiated tibia autograft and prevent nonunion.Patients were 3 cases of osteosarcoma,1 of Ewing's sarcoma,and 1 of relapse of Langerhans's cell histocytosis in tibia.The length of resect bone was 14.0-20.0 cm (mean,17.2cm),constitute of 2 osteoarticular resections and 3 intercalary resections.The method of inactivation of bone segment was intraoperative extracorporal irradiation.Regular followed-up were done postoperative.The X-ray and CT were applied to observe the function of affect limb.The bone union time and complication were record.Results All patients were followed-up of 12-18 months (mean,14.2 months).Eight ends of irradiated-host bone in 5 patients healed completely in 7.8 (6-10) months postoperative.The region of ends were covered by periosteum and showed excellent osteogenic power.There was no leg length discrepancy occurred in patients who received intercalary inactivation because of the preservation of growth plate.But the other 2 osteoarticular inactivated patients suffered leg length discrepancy of 1.0 cm and 1.5 cm respectively because of the inactivation of growth plate.At the follow-up of 12 months post-operation,the mean MTSS of affect limb was 25.8(22-28),and the mean of MTSS％ was 86％(73％-93％).Conclusion Transplantation of pedicled vascularized fibular periosteum can promote effectively healing of irradiated tibia bone after replantation in Children,with simple operation and less complications.

Objective: To understand the trends in malnutrition among primary and middle school students of Han nationality in Hainan Province from 2005 to 2019, so as to provide a basis for improving nutrition intervention measures for children and adolescents. Methods: A sample of 32 949 Han nationality primary and middle school students aged 7-18 years old in Hainan Province were investigated in 2005, 2010, 2014 and 2019 based on national survey on student physical fitness and health. The Malnutrition Screening Standard of Schoolage Children and Adolescents was used to screen malnutrition. Statistical analysis was performed using the χ2 test and the χ2trend test. Results: In the four surveys conducted during 2005 to 2019, the prevalence of malnutrition among primary and middle school students were 22.12%, 18.80%, 15.89% and 9.56%, respectively, with an increase of -12.56% and an average annual increase of -5.82%. The decreasing trend of malnutrition by year was statistically significant (χ2trend=600.72, P<0.01), and the proportion of emaciation type was the highest (8.87%-20.15%). The detection rates of malnutrition among all students aged 7 to 18 showed a decreasing trend from 2005 to 2019 (χ2trend=56.44, 60.04, 61.48, 42.49, 51.81, 50.81, 72.86, 101.34, 86.38, 24.81, 17.72, 10.38, P<0.01). From 2005 to 2019, the detection rates of malnutrition in boys were higher than that of girls (in 4 surveys), and that in rural students from 2005 to 2014 of 3 surveys were higher than that in towns (χ2=92.07, 35.16, 25.29, 29.98; 64.35, 4.26, 6.32, P<0.05). Conclusions The malnutrition of Han nationality primary and middle school students aged 7-18 years in Hainan Province show a trend of improvement year by year from 2005 to 2019, despite the overall high detection rate. Wasting is the most common type of malnutrition. The epidemic of malnutrition varies by age, sex and areas. Further targeted measures should be taken to strengthen intervention in the diet of primary and middle school students, to improve the nutritional status of children and adolescents.

OBJECTIVE: To explore the clinical and genetic characteristics of eight children with Primary hypertrophic cardiomyopathy (HCM). METHODS: Eight children with HCM admitted to the Department of Cardiology of Henan Children's Hospital from January 2018 to December 2021 were selected as the study subjects. Clinical data of the children were collected. Whole exome sequencing was carried out on two children, and trio whole exome sequencing was carried out on the remainder 6 children. Sanger sequencing was used to verify the candidate variants in the children and their parents, and the pathogenicity of the variants was evaluated based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). RESULTS: The patients had included 5 males and 3 females, with their ages ranging from 5 to 13 years old. The average age of diagnosis was (7.87 ± 4.8) years old, and the cardiac phenotype showed non-obstructive HCM in all of the patients. WES has identified variants of the MYH7 gene in 4 children, including c.2155C>T (p.Arg719Trp), c.1208G>A (p.Arg403Gln), c.1358G>A (p.Arg453His), and c.1498G>A (p.Glu500Lys). Based on the guidelines from the ACMG, the first 3 variants were classified as pathogenic, while c.1498G>A (p.Glu500Lys) was classified as likely pathogenic (PM1+PM2_Supporting+PM6+PP3), which was also unreported previously. The remaining four children had all harbored maternal variants, including MYL2: c.173G>A (p.Arg58Gln; classified as pathogenic), TPM1: c.574G>A (p.Glu192Lys) and ACTC1: c.301G>A (p.Glu101Lys)(both were classified as likely pathogenic), and MYBPC3: c.146T>G (p.Ile49Ser; classified as variant of uncertain significance). Seven children were treated with 0.5 ~ 3 mg/(kg·d) propranolol, and their symptoms had improved significantly. They were followed up until September 30, 2022 without further cardiac event. CONCLUSION Genetic testing can clarify the molecular basis for unexplained cardiomyopathy and provide a basis for clinical diagnosis and genetic counseling. Discovery of the c.1498G>A (p.Glu500Lys) variant has also expanded the spectrum of MYH7 gene mutations underlying HCM.
Female ; Male ; Humans ; Child ; Child, Preschool ; Adolescent ; Cytoskeletal Proteins ; Family ; Genetic Counseling ; Genetic Testing ; Cardiomyopathy, Hypertrophic/genetics*

OBJECTIVE: To explore the clinical and genetic characteristics of five children with Catecholaminergic polymorphic ventricular tachycardia (CPVT). METHODS: Five children with clinical manifestations consistent with CPVT admitted to the Department of Cardiology of Children's Hospital Affiliated to Zhengzhou University from November 2019 to November 2021 were selected as the study subjects. Their clinical data were collected. Potential variants were detected by whole exome sequencing, and Sanger sequencing was used to verify the candidate variants. All patients were treated with β-blocker propranolol and followed up. RESULTS: All patients had developed the disease during exercise and presented with syncope as the initial clinical manifestation. Electrocardiogram showed sinus bradycardia. The first onset age of the 5 patients were (10.4 ± 2.19) years, and the time of delayed diagnosis was (1.6 ± 2.19) years. All of the children were found to harbor de novo heterozygous missense variants of the RYR2 gene, including c.6916G>A (p.V2306I), c.527G>C (p.R176P), c.12271G>A (p.A4091T), c.506G>T (p.R169L) and c.6817G>A (p.G2273R). Among these, c.527G>C (p.R176P) and c.6817G>A (p.G2273R) were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.527G>C (p.R176P) was classified as a pathogenic variant (PS2+PM1+PM2_Supporting+PM5+PP3+PP4), and the c.6817G>A (p.G2273R) was classified as a likely pathogenic variant (PS2+PM2_Supporting+PP3+PP4). The symptoms of all children were significantly improved with the propranolol treatment, and none has developed syncope during the follow up. CONCLUSION Discovery of the c.527G>C (p.R176P) and c.6817G>A (p.G2273R) variants has expanded the mutational spectrum of the RYR2 gene. Genetic testing of CPVT patients can clarify the cause of the disease and provide a reference for their genetic counseling.
Child ; Humans ; Mutation ; Propranolol ; Ryanodine Receptor Calcium Release Channel/genetics* ; Syncope ; Tachycardia, Ventricular/diagnosis* ; United States