BACKGROUND: Compared to those original viruses systems, adeasy adenovirus, a recombinant adenoviral system widely used in recent years, based on viruses with a deletion of both El and E3, reported by T.C. He in 1998, is an improved one. It simplifies the generation and production of such viruses and expedite the process of generating and testing recombinant adenoviruses using homologous recombination in bacteria rather than in eukaryotic cells. Moreover, it can be conveniently followed with the aid of green fluorescent protein encoded by EGFP gene incorporated into the viral backbone.OBJECTIVE: To construct the recombinant adenovirus and to evaluate them by transfect them to mesenchymal stem cells (MSCs)and detect the expression of target gene hlGF-I at gene and protein levels.DESIGN: Repetitive measurement wail.SETTING: The Institute of Pediatric Research, Chongqing University of Medical Science.METHODS: The study was performed at the Institute of Pediatric Research, Chongqing University of Medical Science from November 2004 to March 2005. After the amplification of truncated hlGF-1 gene from pcDNA3.l-hlGF-I by polymerase chain reaction (PCR), the gene fragment was inserted into the shuttle plasmid pAdtrack-CMV for homologous recombination with backbone plasmid pAdeasy-I in bacteria BJ5183 to get adenovirus.Ad-hlGF-1. The high titer adenovirus supernatant was obtained by repeated transducing of HEK 293 cells by adenovirus harvested after confirmation of the adenovirus structure. As target cells,MSCs were infected with adenovirus earned target gene, hIGF-1, to determine the expression of hlGF-1 gene.MAIN OUTCOME MEASURES: ① The construction of recombinant adenovirus vector;② the expression of target gene hIGF-1 in HEK 293 cells and the proper multiplicity of infection (MOI); ③ hIGF-1 gene expression in MSCs.RESULTS: The adenovirus vector based on adeasy system was constructed successfully and the Ad-hlGF transducing was successfully or efficiently expressed in MSCs cells. The ideal expression of harvested recombinant adenovirus in MSCs was detected by fluorescence microscope, RT-PCR, immunocytochemistry, and Western Blot.CONCLUSION: Adenovirus vector is an effective vector tools for gene expression and wansfection of MSCs. MSCs transduced with Ad-hIGF-1 maybe another option to gene-modified seed cells for articular cartilage tissue engineering.

Asphyxia ; etiology ; Child ; Dysostoses ; complications ; Female ; Humans ; Osteochondrodysplasias ; Thorax ; abnormalities

BACKGROUND:In high-intensity exhaustive exercise process, the body must bear the exercise intensity decreasing splanchnic blood flow“ischemia”, at the same time, along with the movement of energy and material consumption, metabolite accumulation and oxidative stress in the body cause pathological damage, leading to a decline in exercise capacity. Thus, what is the impact on kidney filtration barrier? How to adapt to the change of renal tissue? Houttuynia cordata has the functions of heat clearing and detoxifying, dieresis for treating strangurtia, hemostatic, expel ing phlegm to arrest coughing and analgesia, if it has a protective effect on the renal injury caused by acute exhaustive exercise and its mechanism has not been reported in the literature. OBJECTIVE:To explore the effect of acute exhaustive exercise on kidney filtration barrier in rats and the intervention effect of Houttuynia cordata.METHODS:After resting and watching for 3 days, Sprague-Dawley rats received adaptive running for 15 minutes at a speed of 10 m/min on a 0° treadmil . A total of 24 rats, which can finish the running, were selected. They were divided into normal control group, exhaustive exercise group and dosed exhaustive exercise group according to the weight of layer (n=8). Rats in the exhaustive exercise group and dosed exhaustive exercise group on the 10° treadmil received once exhaustive exercise. Dosed exhaustive exercise group received intraperitoneal injection of sodium houttuyfonate 10 mL/kg at 30 minutes before exercises. The normal control group did not do any exercise. RESULTS AND CONCLUSION:Compared with the normal control group, serum urea nitrogen, serum creatinine, urine protein content, malondialdehyde concentration, renal cellapoptosis and apoptosis index were significantly increased, but nitric oxide content and nitric oxide synthase activity in the renal tissue were significantly deceased in the exhaustive exercise group (P<0.05 or 0.01). Glomerular filtration epithelial cells, the kidney filtration barrier of basement membrane and podocyte damage were obvious, showing abundant cellapoptosis, occasional y necrosis. Compared with the exhaustive exercise group, urine protein content, serum creatinine, malondialdehyde concentration, renal cellapoptosis and apoptosis index were significantly reduced, but nitric oxide content and nitric oxide synthase activity were significantly increased in the dosed exhaustive exercise group (P<0.05). No obvious pathological changes were detected, but apoptosis was visible. These findings confirmed that houttuynine made a reduction in renal cellinjury induced by exhaustive exercise and possibly significantly reduced apoptosis, increased nitric oxide synthase content, decreased malonaldehyde, and apparently increased superoxide dismutase activity, and final y protected injured renal tissue induced by exhaustive exercise.

Objective:To explore the feasibility and influencing factors of E-cervix cervical elasticity analysis technology in analyzing normal cervical function during non pregnancy.Methods:213 women who underwent vaginal ultrasound examination in the Ultrasound Department of Foshan Maternal and Child Health Hospital from May 2019 to November 2019 were selected as the research objects. Taking the median sagittal section of the cervix as the initial section, the E-cervix technology software package was started to automatically obtain the elastic contrast index (ECI), hardness ratio (HR), cervical strain rate (IOS), cervical strain rate (EOS), cervical strain ratio (IOS/EOS) and cervical length (CL). The relationship among age, menstrual cycle, BMI index, birth history, delivery mode and elastic parameters were compared.Results:There was no correlation between the elastic parameters and age, and there was no significant difference among different age groups ( P>0.05); there was no significant difference in the elastic parameters of cervical tissue in menstrual period, proliferative period and secretory period ( P>0.05); there was no significant difference in the elastic parameters of underweight, normal and overweight ( P>0.05); CL was positively correlated with body mass index (BMI) ( r=0.225, P<0.05), there was no correlation between other parameters and BMI ( P>0.05); there was no significant difference between the elastic parameters of patients with and without birth history ( P>0.05); the CL of women with cesarean section [(34.22±4.96)mm] was higher than that of women with natural birth [(29.03±4.14)mm] ( P<0.05), and the other parameters had no statistical significance ( P>0.05). Conclusions:The elastic parameters of cervix obtained by E-cervix technique are not affected by age, BMI, menstrual period, reproductive history and delivery mode, and can be used for quantitative evaluation of cervical function.

0.05).There were no signifi-cant relation between the expressions of Fas,FasL and bcl-2and IL-8.Conclusion The abnormal expressions of Fas,FasL,bcl-2may play an important role in the pathogenesis of LN.The expression of Fas and FasL may serve as active indexes for SLE and LN.The low level of IL-8may be related to the dysfunction of immunity and needs further research.

Objective To assess the clinical threatment results of pilon fractures managed with arthroscopyassisted minimally invasive percutaneous plate osteosynthesis (MIPPO).Methods 33 patients with pilon fractures were classified into 3 groups according to the Ruedi-Allgower classification:type Ⅰ in 26 cases,type Ⅱ in 5 cases,type Ⅲ in 2 cases,including 29 males and 4 females,aged 22 to 51 years,mean 31.5 years of age.All patients were treated with arthroscopy-assisted MIPPO with the postoperative follow-up time of 12 to 84 months.Results The clinical surgery efficacy according to Mazur's criterion was evaluated as excellent in 22 cases,good in 8 cases,fair in 3 cases.The excellent and good rate was 90.9％.Conclusion Arthroscopy-assisted MIPPO surgical treatment is an effective method for Pilon fractures with the advantages of good healing,minimal trauma and less complications,it is worthy of clinical application.

Batch and continuous fermentation were adopted to investigate the effect of specific growth rate and amino acid components on RNA accumulation in Candida tropicalis ATCC 20408 in fermentation medium ( FM), yeast peptone dextrose medium (YPD), molasses fermentation medium ( MFM) and FM without corn steep liquor. The data showed that obvious differences in intracellular RNA accumulation were observed at different cell growth phases in bath fermentation prosess, and RNA level reached 11. 8% (g-RNA /g-DCW) during exponential phase, and only 6.9% during stationary phases. It was also found that intracellular RNA accumulation increased with the increase of specific growth rate in continue fermentation prosess, and the highest RNA level reached 15. 6% with the glucose conversion rate of 42. 8% at the dilution rate of 0. 5 h-1. Furthermore, the data showed that RNA lever was notably increased in batch fermentation process when amino acids or peptone was added into the fermentation medium containing no corn steep liquor. Taken together, it was reported for the first time that specific growth rate and amino acid components plays a leading role on the intracellular RNA accumulation in C. tropica lis, and specific growth rate is more important.

Objective To investigate and analyze the iodine nutritional status of school-age children six months after implementation of a new standard of iodized salts in northern area of Jiangsu Province,and to provide a basis for control of iodine deficiency disorders.Methods According to the water iodine content,northern area of Jiangsu Province was divided into low,moderate and high iodine areas (the water iodine contents were ＜ 10,l0-150 and ＞ 150 μg/L,respectively).Two towns were selected in each area; one elementary school was selected in each town; and 120 children aged 8-10 were selected in each school.The samples of urine,household salt,and water in each town were collected.Urinary iodine content was tested by arsenic cerium catalytic spectrophotometry;iodine content of household salt sample was determined by direct titration; iodine content of water sample was tested by cerium sulfate catalytic spectrophotometry.Results In low,moderate and high water iodine areas,240,242 and 242 urine samples of 8-10 years old children were tested; urinary iodine median was 135.49,183.62 and 448.99 μg/L,respectively,and the difference between groups was statistically significant (x2 =165.56,P ＜ 0.05).In low,moderate and high water iodine areas,100,131 and 117 household salt samples were tested,and the median of salt iodine was 21.88,22.01 and ＜ 5 mg/kg,respectively.Water samples collected in low,moderate and high iodine areas were 12,14 and 10 copies,respectively,and the median of water iodine was 8.89,38.52 and 189.25 μg/L,respectively,and the difference between groups was statistically significant (x2 =22.27,P ＜ 0.05).Conclusions The iodine nutritional condition of 8-10 years old children in low,moderate water iodine areas in northern area of Jiangsu Province is appropriate.The new standard is suitable for iodine nutritional needs in the area.The iodine nutritional condition in high water iodine area is excess,iodine intake should be reduced.

OBJECTIVETo construct recombinant lentivirus that stably suppresses phospholipase C (PLC) gamma1 expression in human colorectal carcinoma LoVo cells to obtain LoVo cell lines deficient in PLC gamma1 for investigation of the role of PLC gamma1 gene.

METHODSRecombinant lentivirus producing PLC gamma1 siRNA were constructed to infect LoVo cells, and the stably transduced cells were selected with blasticidin. The protein and mRNA expression of PLC gamma1 was examined by Western blotting and RT-PCR, and the effect of the lentivirus on cell apoptosis was analyzed by flow cytometry.

RESULTS AND CONCLUSIONPLC gamma1 siRNA significantly suppressed PLC gamma1 expression in LoVo cells, suggesting high efficiency of gene silencing induced by the siRNA produced by the recombinant lentivirus. Concomitantly, cell apoptosis induced by 5-FU was significantly increased.

Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; genetics ; Blotting, Western ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; pathology ; DNA, Recombinant ; genetics ; Fluorouracil ; pharmacology ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Phospholipase C gamma ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective To explore a method for isolation and culture of human epidermal stem cells. Methods Epidermis was obtained by digesting human foreskin with Dispase Ⅱ and Trypsin-EDTA.After suspension on the epidermal stem cell medium (ESCM), these single epidermis cells were inoculated onto human collagen Ⅳ-coated flasks and cultured at 37 ℃ in a humidified atmosphere containing 5% CO_2 for 10 min. The nonadherent cells were rinsed off 10 min after inoculation, and the adherent cells continued to be cultured after enriching and abstraction by type Ⅳ collagen. The cell growth was observed through inverted microscope, and the cell cloning efficiency and time of clone sustain were also detected. Immunocytochemistry was used to observe the expression of ?_1-integrin and keratin 19(K19). Keratinocytes were served as controls. Results It was revealed by histological observation that colonies were formed 24 hours after inoculation. The isolated and cultured cell cloning efficiency was higher and the time of clone sustain was longer than that of the control group. Positive expression of ?_1-integrin and K19 of cultured cells was detected by immunocytochemistry. Conclusion Adult epidermal stem cells could be successfully isolated and cultured by adhension with type Ⅳ collagen and culture with ESCM.