[ ABSTRACT] AIM:To establish a perforated whole-cell patch-clamp technique withβ-escin to record L-type cal-cium current (ICa,L) in osteoblasts.METHODS:ROS 17/2.8 is a rat osteoblast-like osteosarcoma cell line.β-escin was applied to the pipette solution to permeabilize the cell membrane and the perforated patch recording mode was obtained. RESULTS:β-escin at concentration of 50μmol/L easily permeabilized the cell membrane and obtained a perforated patch recording mode in 2~7 min.This technique prevented ICa,L rundown and preserved cytoplasmic signaling pathways.CON-CLUSION:β-escin may be used as an alternative ionophore for perforated patch-clamp studies in osteoblasts and results in minimal rundown that could facilitate recordings of ICa,L in osteoblasts.

Objective To approach the value of CT double windows technique in the location of lung X-ray knife.Methods The varying window width(W),window center(C) of peripheral type carcinoma of lung and their error were deltermined.The best double window was chosen and the clinical matherial was analysed.Results (1)The biggest error among different widow widths measuring foci was 7 mm.(2)The best double windows in the location of lung X-ray knife were W1 120~180,C1-700,W2 350,C2 15.(3)34 cases of lung carcinoma were observed with double windows,the outline of foci was contiuous and clear in the 15 cases of peripheral type carcinoma of lung and 8 cases of central type carcinoma of lung without hilum infiltration(67.6%),the inner outline of foci was disappear but could be remedied by mediastinum window in 11 cases of central type carcinoma of lung with hilum infiltration. Conclusion The double widows technique is provied with accuracy and without lack fidelity,and could reflect more condition of peritumour infiltration,it is clear superior to single display technique.

Objective To study the ultrastructural changes of hypertrophic scars and animal wound healing models induced by radionuclide 90 Sr exposure and to get the most effective dosage and time in the prevention and treatment of scars. Methods The clinical hypertrophic scars and animal wound models were exposed using 90 Sr applicator in this study. The exposure doses were 200 800 cGy and 200 4 000 cGy. Then the fibroblastic ultrastructure of the tissues from the experimental and control groups were observed with transmission electron microscope. Results Compared with the control groups, capillaries and fibroblasts obviously increased in small and medium doses (200 600 cGy) groups and fibroblastic function was activated. The fibroblasts decreased and fibroblastic function was inhibited in large dose (800 2 000 cGy) groups. Conclusions Small and medium dose of 90 Sr can accelerate wound healing, and can therefore be used in the treatment of early wounds (2 3 days after wounded) ; large dose of 90 Sr can prevent scars from hyperplasia, and can be used in the wounds of the first week after operation; 1 000 2 000 cGy 90 Sr can cure the old hypertrophic scars or keloids; It is useless that 90 Sr exposes before operation for prevention of scars.The most effective method to prevent scars from hyperplasia is large dose of 90 Sr exposure after operation.

Objective To investigate the CT manifestations and differential diagnosis of benignity and malignancy of appendiceal mucocele.Methods The clinical and imaging data of 31 pathologically confirmed appendiceal mucocele were analyzed retrospectively,in which simple mucocele and mucinous cystadenoma were devided into benign group;low grade appendiceal mucinous neoplasm(LAMN) was devided into borderline group;mucinous cystadenocarcinoma was devided into malignant group.The CT manifestations were analyzed and compared statisticallyl in size(long diameter and short diameter), cyst wall(thickness, evenness, smoothness), intra-cystic contents(mural nodules, septum, calcification, intra-cystic fliud density), periappendiceal fat stranding, intraperitoneal free fluid and pseudomyxoma peritonei.Sensitivity and specificity values were calculated if single or multiple CT manifestations were used.Results Of the 31 cases of mucocele of the appendix, 11 of them were in benign group (including 8 cases of simple mucocele and 3 cases of mucinous cystadenoma);14 cases in borderline group (LAMN) and 6 cases in malignant group (mucinous cystadenocarcinoma).CT findings including evenness and smoothness of cyst wall, mural nodule, intraperitoneal free fluid and pseudomyxoma peritonei showed statistically significant difference among three groups (P<0.05).When combined with any four of the five signs, the sensitivity and specificity for diagnosing malignant mucocele were 83% and 88% respectively.But no significantly difference were found among the three groups(P>0.05) in long diameter and short diameter of mucocele, attenuation of intraluminal contents, maximal wall thickness, calcifications, internal septa and periappendiceal fat stranding.

Objective To observe the distribution of the neuropeptides in mouse sublingual gland. Method Using immunohistochemical ABC method. Results The striated duct cells in the sublingual gland showed somatostatin and calcitonin gene related peptide immunoreactivity positive which were in the cytoplasma, but the nucleus negative.Conclusion The striated duct cells in mouse sublingual gland also contained neuropetides; which may play an important role in regulating acinar secretion and blood supply.

Objective To analyze the false-positive results of Treponema pallidum antibody caused by 3 different assay in comparison with Treponema pallidum hemagglutination assay (TPHA).Methods Research group included 3957 clinically asymptomatic syphilis patients,and control group was 344 outpatients with sex-transmitted diseases (STD).The serum samples from the patients who were TPHA-positive were tested in parallel by enzymeimmunoassay (EIA) and syphilis toluidine red untreated serum test (TRUST).Western blot (WB) was performed as confirmatory test.Results In the clinically asymptomatic patients,60 were TPHA-positive.Among them 57 were confirmed by western blot assay,and 1 was false-positive and 2 were borderline in WB.Of the 60 TPHA-positive patients,53 were positive in EIA and 23 were positive in TRUST.In STD patients 40 were TPHA,WB and EIA-positive but 32 were TRUST-positive.Conclusions The results of TPHA and EIA were consistent for diagnosis of syphilis patients who may suffer from previous or latent infection.

Objective To evaluate the accuracy and clinical efficacy of low field phase-contrast magnetic resonance angiography(PC-MRA),CTA and 3D-DSA in the diagnosis of cerebrovascular disease.Methods 70 patients with clinically suspected cerebrovascular diseases were examined with PC-MRA,CTA and 3D-DSA.The sensitivity in detecting the cerebrovascular disease with these three methods were compared.Results Fifty-one patients were detected with cerebrovascular diseases in seventy patients by three imaging methods,including 36 aneurysms in 33 cases and 18 arteriovenous amlformations.33,34 and 35 aneurysms,17,16 and 18 arteriovenous malformations were detected by PC-MRA , CTA and 3D-DSA respectively , the sensitivity were 91.67% and 94.44%,94.44% and 88.89%,97.22% and 100% respectively.There were no significant difference in the sensitivity of detecting aneurysm and arteriovenous malformation between these three imaging methods(P＞0.05).Conclusion PC-MRA,CTA and 3D-DSA are no of difference in diagnosis of cerebrovascular malformation.

Objective To investigate the application of peroneal artery chain perforator flap in repairing skin and soft tissue defects of ankle and lower leg and the corresponding clinical effect.Methods From 2005 to 2010,19 cases of skin and soft tissue'defects of the ankle and lower leg were treated by using peroneal artery chain perforator flaps.The sites of peroneal artery branches were detected by using Doppler flowmeter before operation.Supplied by the chain form of terminal branches of the peroneal artery,the cutaneous fulcrum straightly nearest to the wound was selected as the rotation point to design the flap,with area ranging from 25 cm × 8 cm to 16 cm × 6 cm.Results The follow-up for 1.5- 42 months revealed that all cases of flap survived with satisfactory shape,except for one case suffered from distal skin flap necrosis.Conclusions The peroneal artery chain perforator flap with unsteady peduncle has no restrictions of peduncle sites and can be cut flexibly.It is a perfect method in repair of soft tissue defect of ankle and lower leg.

Objective To investigate the effects of the transient receptor potential V6 (TRPV6) gene silencing on the proliferation and apoptosis of trophoblasts HTR-8/SVneo cells.Methods siRNA sequences targeting the TRPV6 gene were constructed and then transfected into HTR-8/SVneo cells mediated by liposome.The cells were divided three groups,including blank control (add the reagent of transfenction),negative control groups (transfecting nonspecific siRNA) and experimental groups (transfecting TRPV6-siRNA).Those cells in every group were collected at 24,48,72 hours after transfecting.The expression levels of TRPV6 mRNA were detected by reverse transcription (RT) PCR at different times after transfecting.The effects of siRNA on the proliferation and apoptosis of the cells were assayed by methyl thiagolyl tetragolium (MTT) and flow cytometry at different times after transfecting.Results siRNA TRPV6 transfection could inhibit the expression of TRPV6 mRNA in the HTR-8/SVneo cells.The expression was decreased with the extension of time,by 0.72 ± 0.02,0.54 ± 0.02 and 0.29 ± 0.01 after 12,48 and 72 hours of siRNA transfection as compared with the blank control and the negative control groups (P ＜0.01).The rates of proliferation inhibition were (19.29 ± 1.23) ％,(32.12 ± 1.35) ％ and (46.51 ±1.42) ％ at 24,48 and 72 hours respectively when compared with the blank control (2.12 ± 0.03)％,(2.42 ± 0.02) ％,(3.13 ± 0.04) ％ and the negative control groups (2.37 ± 0.01) ％,(2.61 ± 0.05) ％,(2.93 ± 0.03) ％ (P ＜ 0.01).The apoptosis rates of HTR-8/SVneo cells was 16.21％ at 48 hours after transfected with siRNA TRPV6,which were significantly higher than 3.27％ in the blank control and 5.34％ in the negative control groups (P ＜ 0.05).Conclusion Silenceing of TRPV6 genen could inhibit the proliferation and increase the apoptosis of extravillous trophoblas of human placenta.

Objective To investigate the effects of pregnancy complicated with diabetes on the insulin sensitivity of offspring during their early childhood. Methods Offspring of diabetic mothers(ODM) and of non-diabetic mothers(ONDM) aged 1 month to 24 months were recruited into this prospective cohort study and followed up for two years. Body weight and body length were measured at 2, 4, 6, 8, 10, 12, 18 and 24 months of age respectively, and body mass index (BMI) were calculated. Fasting plasma glucose and fasting serum insulin levels were measured on the following-up day at 6, 12 and 24 months of age and insulin sensitivity index (ISI) was calculated. Homeostasis model assessment was used to calculate the insulin resistance (HOMA-IR). Insulin sensitivity was evaluated by fasting serum insulin, ISI and HOMA-IR. The difference of insulin sensitivity between ODM and ONDM group were examined by analysis of covariance adjusted by gender, gestational age,birth weight and BMI. Results Six hundred and five babies including ninety ODM and five hundred and eleven ONDM met the inclusion criteria. There were no differences in gender, gestational age,birth-weight/height between the two groups(P＞0. 05). ODM were heavier and higher than ONDM at each measure point during early childhood, but there were statistical differences at the age of 2, 4 and 6 months only (P＜0. 05). And the BMI at age of 2 and 4 months of ODM were higher than those of ONDM(P＜0.05). The number of baby who accepted the measurement of fasting plasma glucose and fasting serum insulin levels at 6, 12 and 24 months of age was 276 cases, 273 cases and 56 cases respectively. The fasting serum insulin of ODM (logarithmically transformed) were 0. 95±0. 30,0. 89±0. 34 and 0. 90±0. 27, which were higher than those of ONDM (0. 70±0. 45, 0. 73±0. 35 and 0. 67±0. 30) (t=9. 58, 5.01 and 6. 11, P＜0.05); HOMA-IR (logarithmically transformed) were 0. 34±0. 33, 0. 27±0. 36 and 0. 27±0. 31, which were higher than those of ONDM also(0.08±0. 46,0. 10±0. 36 and 0. 03 ± 0.33) (t= 9. 55, 4. 79 and 5. 06, P＜0.05); ISI(natural logarithmically transformed) were -3.87±0. 75, -3.73±0. 81 and -3. 73±0. 71, which were lower than those of ONDM(-3.29±1.05, -3.35±0.84 and -3.18±0. 77) (t=9.20, 4. 90 and 5.06, P＜0.05).There were differences in feeding characteristics of ODM between insulin sensitive subgroup [40. 9%(9/22) breast-feeding] and insulin insensitive subgroup [16.67 % (12/72) breast-feeding] (x2 = 7.02,P=0. 03). Conclusions Pregnancy complicated with diabetes has adverse effects on the offspring insulin sensitivity during their early childhood, and affects the early growth and development of them.Breast-feeding might decrease insulin resistance in babies.