Objective To investigate the structural and ultrastructural changes of the skin induced by micro-plasma radio-frequency technology,and to preliminarily discuss this novel technology mechanism.Methods Thirty guinea pigs were enrolled and randomly divided into three groups.They were radiated by different dose parameters 40 W,10 kJ; 60 W,10 kJ and 80 W,10 kJ.Every guinea pig's back was divided into two parts which was removed after immediately,one week and one month,and dermatopathology and transmission electron microscopy (TEM) were performed.Results The different dose setting could make different skin change of immediately effect.When dose setting was 40 W,10 kJ,skin showed that epidermal cells were integrity and the superficial layer of dermis collagen tissue was light homogenization.When dose setting was 60 W,10 kJ,epidermal tissue showed focal emergence of fractional shape change and obvious homogenization.When dose setting was 80 W,10 kJ,epidermal showed complete vaporization loss or degeneration necrosis,and dermal superficial and middle layer of collagen tissue showed a large area of homogenization.Skin superficial collagen tissue's structure gradually showed dense and arranged in an orderly manner after one week and markedly thickened and arranged in compact manner after one month.TEM showed that epidermal cells were relatively complete,intercellular structure was normal,but the dermal collagen lost originally normal structure and cell structure disappeared and obviously showed massive apoptosis.A small amount of apoptosis was showed but collagen structure gradually restored after one month.Conclusions The novel micro-plasma radio-frequency has obvious dose effect to skin,and its main target tissues are dermal collagen tissue.It can stimulate skin collagen hyperplasia in certain degree.

Objective To investigate the effect of protein kinase B and its phosphorylation on the apoptosis of MIN6 induced by palmitate.Methods Incubated with different level of palmitate concentration (0 ～ 0.5 mmol/L),MIN6 cells were cultured in high glucose DMEM with or without LY294002 ; the apoptosis of MIN6 cells was detec-ted and quantified with TUNEL technology.Then we inspected the cell ultrastructure through electron microscopy and determined the expression of protein kinase B and its phosphorylation p-PKB (Ser473) by Western blot,fol-lowed by an RT-PCR detection of BAX and BCL-2 expression.Results Apoptosis of MIN6 cells was induced by palmitate in a concentration-dependent correlation and enhanced by LY294002.Palmitate also inhibited phospho-rylation of protein kinase B on Ser473.Finally,we detected a downregulation of BCL-2 mRNA expression and up-regulation of BAX mRNA expression under palmitate-incubated state.Conclusion Palmitate may induce apoptosis of pancreatic β-cells through inhibiting activation of PKB phosphorylation in diabetes.

Medical treatment technology has made a lot of breakthrough because of the rapid development of the medical devices industrial.Minimally-invasive and noninvasive medicine is the development tendency of the future medicine.To adapt to the needs of the future medicine,Chongqing medical university exerted much efforts on training of minimally-invasive and noninvasive medical talents.We explored and practiced in aspects of fixing educational conception and goal,building curriculum and teaching content,constructing professional teaching team and developing practice and innovation abilities in order to fill the void of personnel training in the minimally-invasive and noninvasive medical field.

BACKGROUND: Some studies have showed that after indirect co-culture, bone marrow mesenchymal stem cells can differentiate into myocardial cells and hepatocypte-like cells. OBJECTIVE: To investigate the possibility of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) to differentiate into hepatocytes by co-culture with human hepatocyte line LO2 in vitro.METHODS: Full-term umbilical cord blood samples were obtained sterilely. The UCB-MSCs were isolated by density gradient centrifugation and directly adherence growth, then passaged with trypsin digestion at 80% cell fusion. By utilizing cell culture plate insets with microporous membrane combined with 6-well plate, the LO2-/UCB-MSCs co-culture system was established. UCB-MSCs were plated into the wells of 6-well plate at a density of 1×10~7/L. LO2 cells were plated into the cell culture plate insert at a density of 1×10~5/L. UCB-MSCs were plated in both layers in the control group. Surface markers of adhered cells were detected by flow cytometry. Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. mRNA expression was detected by reverse transcriptase PCR. RESULTS AND CONCLUSION: HUCB MSCs expressed CD44 and CD29 strongly, but CD34 and CD45 were expressed negatively. After 5 days, fusiform-shaped cells were reduced in the co-culture group; while, the time passing by, cells shaped irregular round or polygonal were increased, which were similar to hepatocytes. At 4 weeks after culture, UCB-MSCs were still fusiform-shaped in the control group. At day 5 after culture, alpha fetoprotein mRNA expressed positively, but other expressed negatively in the co-culture group; at day 14 after culture, cytokeratin-19 mRNA and albumin mRNA expressions were observed; moreover, with the time passing by, the expression of albumin mRNA was increased, but the expression of alpha fetoprotein-19 mRNA was decreased. Antigenic expressions in the control group were negative. This suggested that UCB-MSCs could differentiate into hepatocypte-like cells by co-culture with human hepatocyte line LO2 in vitro.

AIM:To investigate the mechanism of lipotoxicity-associated apoptosis mediated by mitochondrial pathway in pancreatic β-cells. METHODS:The pancreatic β-cell line MIN6 cells were incubated respectively in serum-free DMEM medium with or without palmitate (0.1-0.5 mmol/L) for 24 h,followed by Hoechst 33258 staining. The activity of caspase 3 was assayed for determining apoptosis,transmission electron microscope was used for observing mitochondrial structure,and RT-PCR analysis was applied for detection of mRNA expression of bcl-2-associated X protein (bax),B-cell lymphoma 2 (bcl-2),sterol regulatory element binding transcription factor 1c (SREBP1c) and DNA-damage inducible transcript 3 (chop-10). RESULTS:Palmitate induced apoptosis of MIN6 cells and swelling of mitochondria. Palmitate also inhibited mRNA expression of bcl-2,whereas enhanced mRNA expression of bax,SREBP1c and chop-10. CONCLUSION:Palmitate induces apoptosis of pancreatic β-cells by promoting mitochondrial dysfunction,which is associated with abnormal expressions of bax,bcl-2,SREBP1c and chop-10.

Objective To observe and compare the changes of weight,blood pressure,glycemia, blood lipid,serum uric acid,insulin sensitivity and insulin function of laparoscopic sleeve gastrectomy (LSG)for the treatment of type 2 diabetes mellitus(T2DM)with obesity,and analysed mechanism of meta-bolic surgery.Methods Changes of weight,blood pressure,glycemia,blood lipid ,serum uric acid,in-sulin sensitivity and insulin function of 8 type 2 diabetes mellitus(T2DM)with obesity patients undergoing LSG were retrospectively analyzed.Weight,BMI,EWL%,average systolic and diastolic pressure were measured 1,2,3,6 months and fasting plasma glucose (FPG),glycosylated hemoglobin (HbAl),insulin sensitivity parameter:insulin resistance index(HOMA-IR),insulin sensitivity index(ISI),insulin area un-der the curve (AUCI),insulin function parameter:glucose disposition index(DI)were measured 1,6 months after operation and compared with preoperative levels.Meanwhile medication change was observed af-ter operation and compared with preoperation.Results ⑴Weight ,BMI at 1,2,3,and 6 months after surgery showed a decreasing trend and EWL% showed a decreasing trend over time .⑵ FBG、HbA1c showed a decreasing trend after operation.Blood lipid,uric acid of 3 patients with dyslipidemia and hyperu-ricemia decreased to normal 1 month after operation.⑶ HOMA-IR,AUCI except for 1 patient showed a de-creasing trend and ISI was contrary after operation.DI ,trends were different.⑷7 patients stopped all an-tidiabetic drugs 6 months after operation,1 patient injected insulin with good glycemic control.Blood pres-sure of 5 patients with hypertension was normal without antihypertensive drugs 6 months after operation. Conclusions LSG procedure can significantly increase insulin sensitivity of T2DM with obesity,thereby improve glycemia,cholesterol,uric acid and further promot weight loss.

Objective To explore the value of diffusion tensor imaging (DTI) and diffusion tensor tracography (DTT) in assessment of Corticospinal tract (CST) and medial lemniscus (ML) in tumors involving brainstem.Methods A total of 35 cases with pathologically confirmed tumors involving brainstem were collected,and 35 volunteers matched with genders and ages were recruited as the normal group.DTI scanning was performed on all the patients and controls.The damage degrees of CST and ML were evaluated and graded by DTT,and the dysfunction degrees were evaluated for the patients.Spearman correlation was used to statistically analyze the relationships of limb movement,sensory dysfunction and CST and ML damage.Results According to the rating results,normal findings,shifting,edema or infiltration and damage of CST was found in 9,9,11,and 6 cases respectively.They were 8,9,15,3 cases for ML.Motor function was normal in 20 cases,slightly defective in 11 cases,and moderate defective in 4 cases.Sensory function was normal in 21 cases,slightly defective in 6 cases,and moderate defective in 8 cases.The patients' dyskinesia and CST damage degree,sensory dysfunction and ML damage degree were positively correlated (r was 0.786 and 0.686 respectively,P ＜ 0.01).The position relationship among tumor and CST and ML could be well displayed on images.None of the patients showed new symptoms of dysneuria after surgery.Conclusions DTI and DTT technology can be used to evaluate CST and ML damage degree in tumors involving brainstem.They can display the position relationship between tumor and the brainstem CST and ML,which is important in protecting the brainstem fiber tract during operation and evaluating the recovery after the operation.

OBJECTIVETo establish a quantitative HPLC method for determination of protopine, palmatine hydrochloride, bicuculline and tetrahydropalmatine, in Corydalis decumbens.

METHODThe separation was performed on a ZORBAX Eclipse XDB-C18 column (4.6 mm x 150 mm, 5 microm) at a flow rate of 1.0 mL x min(-1) using mixtures of two solvents [A(20 mmol x L(-1) ammonium acetate)-B(acetonitrile)]: with a gradient elution. The column oven temperature was 30 degrees C and the detection wavelength was set at 280 nm.

RESULTThe 4 alkaloids were well separated by this HPLC method. Linearifies of protopine, palmatine hydrochloride, bicuculline and tetrahydropalmatine were good in the ranges of 1.44-46.0 (r = 0.999 4), 1.2640.2 (r = 0.999 8), 1.37-44.0 (r = 0.999 9), and 1.3643.6 mg x L(-1) (r = 0.999 9), respectively. The average recoveries were 98.2% with RSD 2.7% for protopine, 101.9% with RSD 2.5% for palmatine hydrochloride, 102.8% with RSD 3.5% for tetrahydropalmatine, and 98.8% with RSD 3.1% for tetrahydropalmatine.

CONCLUSIONThis method is proved to be convenient, reliable and accurate., and it can be used for quality control of C. decumbens.

Objective To compare changes in signal intensity (SI) of the dentate nucleus (DN),pons and cerebrospinal fluid (CSF) on unenhanced T1-weighted magnetic resonance imaging (MRI) scans after multiple administrations of the linear gadolinium-based contrast agent.Methods Clinical and imaging data of forty-nine patients who underwent at least 4 consecutive enhanced MRI examinations were analyzed retrospectively (unenhanced T1-weighted MRI scans were performed before and after consecutive enhanced MRI examinations).The mean SI of DN,pons and CSF were measured on unenhanced T1-weighted images.The SI ratio of DN to pons was calculated by dividing the SI in the DN by that in the pans,and so was The DN to CSF SI ratio.The difference of SI ratio of DN-to-pons and DN-to-CSF in the same patient before and after repeated contrast-enhanced T1WI were analyzed by paired-samples t tests.Linear regression analysis was used to analyze whether the SI ratios of DN-to-pons,DN-to-CSF were correlated with any clinical factor.Results The SI ratios of DN-to-pons and DN-to-CSF after multiple contrast administrations were higher than those before enhancement,and the differences were statistically significant.The SI ratios of DN-to-pons and DN-to-CSF before and after repeated contrast enhancement were 0.98±0.62,1.05 ±0.54;3.89±0.94,4.97±1.61 (t=-8.494,-4.526,P＜0.001).The SI ratio differences showed a significant positive correlation with the number of previous gadolinium-based contrast material administrations (pons:b=0.006,P=0.003,CSF:b=0.144,P=0.009).There was no correlation with any other clinical factors(P＞0.05).Conclusions This study indicates that serial injections of linear gadolinium-based contrast agent may lead to an increase in SI in the DN.

Objective To fabricate manganese-doped carbon quantum dots(Mn-CDs)@anti-human epididymis protein 4(HE4)monoclonal antibody(Mn-CDs@Anti-HE4 mAb)dual-modal fluorescent-magnetic nanoprobe for ovarian cancer cells targeting imaging,and evaluate its potential on fluorescent imaging and MRL Methods Mn-CDs were synthesized at 150 ℃ with solvothermal method.The average diameter,fluorescent capability and MRI efficiency were determined.The cytotoxicity of Mn-CDs in vitro was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)assay with HO-8910 ovarian cancer stem cells and EA.hy926 human umbilical vein endothelial cells.Mn-CDs@Anti-HE4 mAb was fabricated with condensation reaction and characterized by ultraviolet(UV)absorption spectra.Fluorescence imaging and MRI in vitro was performed for cancer cell-targeting study.One-way analysis of variance and the least significant difference t test were used to analyze the data.Results The Mn-CDs with diameter of(4.64±0.85)nm showed a well-defined spherical morphology.The fluorescent spectra of Mn-CDs exhibited a typical excitation-dependent behavior with an excitation maximum at 360 nm and emission maximum at 440 nm.The T1 relaxation rate was(3.26±0.04)mmol ? L-1 ? s-1.The cytotoxicity tests in vitro showed that the survival rates of HO-8910 cells and EA.hy926 cells were both significantly different after treated with different concentrations of Mn-CDs(F= 1 947.509,260.174,both P<0.05),and there was no cytotoxicity in both HO-8910 cells and EA.hy926 cells at concentrations of MnCDs within 0-2.5 mg/ml(all P>0.05),while the survival rates of the two kinds of cells were descended with the increasing of concentration within 3.0-4.5 mg/ml(P<0.05).Mn-CDs@Anti-HE4 mAb could target HO-8910 cells on fluorescence imaging and MRI.Conclusions Mn-CDs@Anti-HE4 mAb,with good potential on fluorescence imaging,MRI and targeting ability,is successfully synthesized.It may provide a new method for early diagnosis of ovarian cancer.