Objective To explore the high risk factors, pathogenesis and methods of early diagnosis of periventricular leukomalacia(PVL) in premature infants.Methods The history of intrauterine hypoxia-ischemia was investigated in premature infants;TORCH-IgM antibodies in cord blood of premature infants were measured by ELISA; Nitric oxide (NO) levels in their cord blood were determined by nitric acid reducing enzyme means. Results Thirty-nine of 52 premature infants in PVL group had a history of intrauterine hypoxia-ischemia; TORCH-IgM antibody positive rate in cord blood of premature infants in PVL group was significantly higher than that in control group(P

Objective To study the effects on tissue CT number caused by scan protocols.Methods The phantom was repeatedly scanned in different protocols by changing only one of parameters,such as X-ray tube voltage,mAs and recon kernel,while other parameters were ketp unchanged.The CT number of different materials in phantom were measured and analyzed.Results The CT numbers of tissues changed remarkably with the tube voltage and had different relativity for different tissues.The CT numbers had positive correlation with kV for such maierials as polyethyle,lexan,perspex,but for teflon the correlation was negative.The mAs and recon kernel had no effects on CT number.Conclusions The CT number of tissue changes with scanning X-ray tube voltage,so the setting of scan parameters should be taken into account in image diagnosis and radiotherapy.

[Objective]To evaluate the role of mature cumulus cells from oocyte-cumulus complex(OCC)in in-vitro maturation(IVM)and establish a new culture technique which is convenient to carry out.[Methods]The cumulus cells of OCC were cut off and dispersed by 1 mL syringe.The cumulus cells were co-cultured with the immature oocytes retrieved from the COH cycles after they adherent to the bottom of the dish.The immature oocytes were experienced IVM procedures in different culture media.They were divided into 3 groups(the oocytes at germinal vesicle stage from one woman were allotted to the same group randomly).Group 1(solution A):basic culture medium+human follicular fluid(hFF);Group 2(solution B):solution A+ cumulus cells(OCC);Group 3(solution C):solution A+ OCC+ follicle stimulating hormone(FSH)+ epidermal growth factor (EGF).Then,the maturation rate,fertilization rate and formation rate of available embryo were observed.[Results]In 113 treatment cycles,298 immature oocytes were performed IVM with solution A,B,and C.The difference for 24 hour maturation rates among 3 groups wag statistically significant(A:45.2%,B:61.7%,C:78.2%,P＜0.05).There was no statistical difference for 25～48 hour maturation rates and normal fertilization rates of mature oocytes.The differences of cleavage rates and rescued embryo rates between group 1 and 2,group 1 and 3 were statistically significant(P＜0.05).The formation rates of available embryo showed an increasing trend from group 1,2,to 3.[Conclusion]After being dispersed by simply beat upon with syringe and adherent culture,the mature cumulus eells from mature OCCs in COH cycles,together with growth factors in the follicular fluid or extraneously supplemented,could promote the IVM of immature oocyte.

Objective To investigate the effects of estrogen on mismatch repiar gene expression in colonic mucosa in vivo. Methods A total of 42 healthy individuals underwent colonoscopy were enrolled in the study. Half an hour before colonoscopy examination, blood sample was taken for determining the serum estradiol (E2) level. N ormal colonic mucosal tissues determined by naked eye under colonoscopy examination were taken in the right hemi colon to detect HMLH1 and hMSH2 gene expression by semi-quantitative RT-PCR and immunohistochemistry staining. Then the correlation of serum E2 levels with hMLH1 and hMSH2 expression in colonic mucosa was analyzed. Results A bimodal curve was presented for the correlation between serum E2 level in healthy individuals and hMLH1 expression in colonic mucosa. A strong positive correlation of E2 level with hMLH1 expression in normal colonic mucosa was observed when serum E2 level was more than 45 pg/ml (For mRNA, P=0. 003, r=0. 701; for immunohistochemistry positivity index, P=0. 000, r=0. 874).However there was no correlation between E2 level and hMSH2 expression. Conclusion High serum E2 level might increase the hMLH1 gene expression in colonic mucosa in vivo.

In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.
Green Fluorescent Proteins ; HEK293 Cells ; Humans ; Plasmids ; Polyethyleneimine ; Transfection ; methods

Objective The purpose of this study is to construct eukaryotic gene vector of herpes simplex virus type 1 thymidine kinase(HSV1-tk)and to observe the expression of HSV1-tk in lung adenocarcinoma AGZY cell line.Methods The full length HSV1-tk gene was amplified by PCR from plasmid pHSV 106 and was inserted into pMD18-T.The recombinant plasmid was recombined with eukaryotic vector plRES 2-EGFP u-sing gene recombinant technique .HSV1 -tk was transfected into adenocarcinoma AGZY cell line with Lipo-fectamineTM 2 000.Fluorescence microscopy was used to detect the transfection and expression of HSV 1-tk.RT-PCR was used to detect the mRNA levels of HSV 1-tk.The cell proliferation was measured by MTT assay .Re-sults A length of 1 130 bp gene sequence was obtained by PCR .The expressions of HSV 1-tk at mRNA and protein levels were displayed by RT -PCR and Western blot .MTT analysis showed that there were no significant changes cell survival on after transfection .Conclusion The eukaryotic expression vector of HSV 1 -tk report gene is successfully constructed and HSV 1-tk is effectively expressed in transfected AGZY cells .

Objective To evaluate the postoperative pain induced by various therapeutic catheters after abdominal surgery.Methods A prospective study was conducted in patients selected based on the inclusion criteria.The general condition of the patients was recorded,and nutritional risk screening was performed.The indwelling of therapeutic catheters after abdominal surgery were recorded,including urinary catheter,nasogastric tube,peritoneal drainage tube,common bile duct drainage tube,wound drainage tube,central venous catheter and peripherally inserted central catheter.The pain caused by each type of catheters was evaluated using visual analog scale at 24,48 and 72 hours after tube/catheter insertion.Results A total of 157 patients were selected,including 70 males and 87 females,aged (60.5 ± 12.5) years,with a body mass index of (23.8 ± 3.2) kg/m2,and a total nutritional risk rate of 42％.According to visual analog scale scores,the degrees of pain due to the therapeutic catheters,in descending order,were as follows:4.9 ± 1.7 for nasogastric tube,3.6 ± 0.9 for wound drainage tube,3.0 ±0.9 for urinary catheter,2.6 ±0.9 for central venous catheter,2.4 ± 1.0 for peritoneal drainage tube,1.9 ± 0.7 for common bile duct drainage tube,and 1.8 ± 0.8 for peripherally inserted central catheter.The catheter-induced pain accounted for (44.9 ± 14.1)％ of the total pain during the hospital stay.Conclusions Nasogastric tube,wound drainage tube and urinary catheter can increase the pain of patients.It is therefore recommended to remove the indwelling tubes as early as possible if only the removal does not harm the outcome of the patient.

Objective To investigate the efficacy of zoledronic acid in the treatment of senile unstable femoral intertrochanteric fractures with anatomical locking plate. Methods 67 patients were randomly divided into two groups. Five days after the operation, group A received one intravenous injection of 5 mg zoledronic acid, while patients in group B did not receive the injection. The two groups were compared in terms of hospitalization time, complications, limb weight-bearing time, fracture healing time, hip function score after operation, preoperative and postoperative serum calcium and serum ALP, bone mineral density of proximal femur before operation and 1 year after operation. Results There were no statistically significant differences between the two groups in age, type of fracture, hospital stay, partial weight-bearing time, fracture healing time, hip function at 1 month and 1 year after operation, preoperative bone mineral density and blood calcium. But the differences were statistically different in hip function at 3 months after operation , averaged bone mineral density of proximal femur and serum ALP 1 year after operation. Moreover, 5 patients in group A developed muscle pain or fever after intravenous injection of zoledronic acid. Conclusion The locking plate combined with zoledronic acid injection in treatment of elderly patients with unstable femoral intertrochanteric fracture could inhibit bone loss, increase bone mineral density, and accelerate limb function recovery after operation. On the other hand, Zoledronic acid has a high incidence of adverse reaction.

Objective To compare the clinical effect of vascularized fibular flap transplantation and vancomycin sulfate calcium on chronic osteomyelitis of the tibia and bone defects.Methods A total of 35 cases with chronic tibial osteomyelitis and bone defect were involved and divided into group A with 21 cases receiving anastomosis of vascularized fibular flap transplantation and group B with 14 cases recciving vancomycin sulfate calcium.Infection control rate and Enneking score were compared in both groups.Simultaneous detection of two groups preoperative,postoperative 2 weeks,1 month and 3 months of the levels of CRP and PCT were conducted.Results Postoperative CRP and PCT were under control in both groups.No statistical significance was found in the comparison of CRP and PCT preoperative and postoperative 2 weeks,1 month and 3 months.In terms of infection control rate,two groups had no signiticant difference (P ＞ 0.05) and there was statistical significance of the comparison of postoperative Enneking score (P ＜ 0.05).Conclusions The clinical results of the two methods are satisfied.The infection degree and the length of the bone defect are decisive factors in choosing the treatment method and sometimes the combination of two treatment methods may be a better choice.

Objective To investigate the ionic mechanism of remifentanil-induced vaso-relaxation by recording calcium-activated K~+-currents(IKCa)in human arterial smooth muscle cells(MASMCs).Methods Human MASMCs were obtained freshly by the method of enzymolysis.IKCa were separated with 4-AP 5 mmol?L~(-1) and recorded by the whole-cell voltage-clamped technique.The effects of remifentanil(1.2,4.8,19.4 mmol?L~(-1))on amplitude and threshold of activation for IKCa were assessed.Results Remifentanil significantly increased IKCa and had no effect on the threshold of activation for IKCa as compared with the control group(P＜0.05 or 0.01),and the IKCa returned to the baseline level after remifentanil was washed out.Conclusion Remifentanil produces vasodilation by activating IKCa in MASMCs.