BACKGROUND:Under the pathological conditions of urethral defect,stricture and diverticulum induce by trauma,congenital malformation,tumor and operation,autogenous tissue are used for defect repair.OBJECTIVE:To verify the feasibility of urethra constructed by tissue engineering.DESIGN,TIME AND SETTING:An observational experiment was performed at Linbaixin Medical Research Center,Sun Yat-sen University,from January 2005 to January 2006.MATERIALS:Ten New Zealand rabbits weighing 3.0-3.5 kg were used to prepare acellular urethras.Twenty BALB/cA-nude mice weighing 18-20 g,4-6 weeks old,were cultured with extracellular matrix materials in vivo.METHODS:The whole urethra harvested from New Zealand rabbit prepared the extracellular matrix materials by acellular treatment.Half a glans harvested from adult New Zealand rabbit was used for smooth muscle cells proliferation by tissue explant.The corpus spongiosum smooth muscle cells were implanted in acellular matrix materials.Then,they were cultured by subcutaneous embedding in nude mice.There were two groups in the experiment:experimental and control.The complexs of corpus spongiosum smooth muscle cells implanted in acellular matrix materials were embedded in the experimental group,while the simple acellular matrix materials were embedded in the control group.MAIN OUTCOME MEASURES:After 1,2,4,6,8 weeks of embedment,the growth of the complexs were examined by gross,histologic and electron microscopic observation.The expression of smooth muscle cells were identified by immunohistochemistry method.RESULTS:①Under the gross observation,incisions on the back of the nude mice healed well,the diets and activities of the nude mice were normal.In the experimental group,there were thin tissue membranes on the surface of the acellular matrix materials at 1-2 weeks after the embedment.The thickness of tissue membrane increased gradually and there were small blood vessels grown at 4 weeks.The new cells were instead of acellular matrix materials at 6-8 weeks.②Under HE straining,the acellular matrix materials were absorbed.Various cells,most of them were smooth muscle cells,would be instead of the acellular matrix materials.And unequal blood vessels grown in matrix materials,most of them had the trend of blood sinus formation.These results suggested that the complexs formed the structure similar to normal urethra in the nude mice.③Under the electron microscopic observation,there were blood vessels grown in the matrix materials from 4 weeks after embedment.More smooth muscle cells,endothelial cells,and little fibroblast cells,macrophages were observed,as well as some apoptotic smooth muscle cells.④After immunohistochemical staining of ?-smooth-muscle actin antibody,the growth of smooth muscle cells could be observed.CONCLUSION:The corpus cavernosum tissue similar to normal urethra can be constructed by using tissue-engineering technique.

Objective:To investigate the application value of magnetic resonance dynamic contrast enhancement magnetic resonance imaging (DCE-MRI) combined with microRNA-21 (miR-21) and miR-92a in the benign and malignant differentiation of bone tumors and the evaluation of their efficacy.Methods:A total of 120 patients with bone tumors were selected retrospectively from June 2018 to June 2019 in Panyu Hospital of Chinese Medicine, including 52 cases in the benign group and 68 cases in the malignant group. The DCE-MRI dynamic enhancement parameters and tumor tissue miR-21, miR-92a levels were compared between the two groups. The diagnostic value of DCE-MRI, miR-21, miR-92a levels of cancer tissue and their correlation were analyzed. Patients with bone malignant tumors were given comprehensive treatment. Six months after operation, according to the criteria of solid tumor curative effect, they were divided into good curative effect group and poor curative effect group. The DCE-MRI dynamic enhancement parameters [signal enhancement amplitude (SEE), early dynamic enhancement slope value (Slope), centripetal enhancement rate (DER)], miR-21, miR-92a levels of patients with different curative effects were compared.Results:The levels of SEE, Slope, DER and miR-21 a nd miR-92a in the malignant group were higher than those in the benign group ( P<0.05); The area under curve (AUC) of DCE-MRI, miR-21, miR-92a combined in the diagnosis of benign and malignant bone tumors (0.885)>Slope(0.808)>SEE(0.788)>miR-21(0.785)>miR-92a (0.740)>DER(0.660), with sensitivity 80.88%, specificity 88.46%, respectively; the DCE-MRI dynamic enhancement parameters SEE, Slope, DER were positively correlated with miR-21 and miR-92a ( P<0.05); the DCE-MRI dynamic enhancement parameters SEE, Slope, DER and miR-21, miR-92a of patients with good curative effect were lower than those with poor curative effect ( P<0.05). Conclusions:DCE-MRI dynamic enhancement parameters, miR-21, miR-92a levels are abnormally high expression in patients with bone malignant tumors, and combined detection is expected to become an important means to identify benign and malignant bone tumors and evaluate the efficacy.

AIM:To investigate the effect of RNA interference (RNAi)-mediated insulin-like growth factor 1 receptor ( IGF1R) gene silencing on the growth , migration, and invasion of hepatocellular carcinoma cells .METHODS:The most effective siRNA targeting IGF1R gene was designed and screened .After lentiviral expression vector pLVX-shR-NA2-IGF1R carrying the most effective siRNA sequence was constructed , it was transfected into 293T cells and packed into pLVX-shRNA2-IGF1R lentivirus.Huh7 and Hep3B cells were infected with the pLVX-shRNA2-IGF1R lentivirus to screen the positive clone Huh7 cells and Hep3B cells with the lentivirus .These Huh7 cells and Hep3B cells were cultured to ana-lyze the mRNA level of IGF1R, cell proliferation, cell cycle, cell apoptosis, cell migration/invasion, and the protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL.RESULTS:The mRNA expression of IGF1R in Huh7 cells and Hep3B cells with pLVX-shRNA2-IGF1R lentivirus was significantly reduced .The proliferation of these cells was remarkably inhibited , and the number in G 1 phase was increased significantly .The percentages of apop-totic cells were increased markedly , and the number of cell migration/invasion was decreased markedly .The protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL were decreased significantly compared with the blank control group and negative control group .CONCLUSION:The RNAi-mediated IGF1R gene silencing sig-nificantly suppresses the growth and the malignant biological characteristics of Huh 7 cells and Hep3B cells, which may be involved in the reduced protein levels of the above genes induced by down -regulation of IGF1R expression.

OBJECTIVETo explore the impacts on the cognitive level of the children with mental retardation (MR) treated with JIN's three scalp needling therapy and the training for cognitive and perceptual disturbance so as to seek the more effective therapeutic method for the treatment of MR in children. Methods Sixty-six MR children were randomized into a head-point needle-retaining therapy + training group (group A) and a training after acupuncture group (group B). Seven cases and 12 cases were dropped in the two groups respectively. Twenty-six cases accomplished the treatment in the group A and 21 cases in the group B. In the group A, the points on the head were stimulated and the needles were retained (30 min after the general acupuncture, the needles on the body points were removed; and those on the head points were retained for 1 h, including Sishenzhen, Naosanzhen, Zhisanzhen and Niesanzhen). Simultaneously, the training for the cognitive perceptual disturbance was combined. In the group B, 1 h after needle retaining at the body points and head points, all the needles were removed; the training for the cognitive perceptual disturbance was followed. The treatment was given once a day, and the treatment for 3 months was required. Before and after treatment, the Wechsler intelligence scale for children (WISC) was used for evaluation and observation of verbal intelligence quotient (VIQ) , performance intelligence quotient (PIQ) and full-scale intelligence quotient (TFIQ) and score changes of 11 items such as information, picture vocabu; lary, arithmetic, picture generalization, comprehension, etc.

RESULTSAfter treatment, FIQ, VIQ and PIQ scores were different significantly as compared with those before treatment (all P<0. 01). In the group A, the results of picture vocabulary, animal egg laying, maze, block design and geometric figure were all improved significantly (all P<0. 05). In the group B, the results of information, comprehension, block design and geometric figure were all improved significantly (all P<0. 05). After treatment, concerning the value difference in FIQ and PIQ of the two groups; the changes in the group A were more significant (both P<0. 01). After treatment, the results of picture vocabulary and maze were improved significantly in the group A as compared with the group B (both P<. 01). The IQ categories changed apparently after treatment in the two groups, toward the higher level in tendency generally, but without significant difference (both P>0. 05).

CONCLUSIONThe simultaneous treatment with head point retaining of JIN's three needling therapy and the training for cognitive and perceptual disturbance obviously improves children patients' verbal comprehension, expression ability, hand-eye coordination ability, attention, logical reasoning ability and visual perception. The efficacy is better than that in the treatment of the training after acupuncture.

Acupuncture Therapy ; Child ; Child, Preschool ; Cognition ; Female ; Humans ; Intellectual Disability ; psychology ; therapy ; Intelligence Tests ; Male ; Perception ; Scalp ; Treatment Outcome

A high throughput screening method based on QuEChERS purification and stable isotope dilution-liquid chromatography coupled to high resolution time-of-flight mass spectrometry was developed for the simultaneous rapid determination of 86 kinds of glucocorticoids (GCs) in cosmetics.The analytes were extracted by acetonitrile, and then the extracts were purified using an improved QuEChERS method.The chromatographic separation was performed on a novel multiple chromatographic retention mechanisms column of Poroshell 120 PFP (100 mm × 2.1 mm, 2.7 μm) with gradient elution using 0.2% (V/V) acetic acid and acetonitrile as mobile phase.The accurate mass database of parent ions and mass spectra library of fragment ions of 86 GCs were established under positive ionization mode with electrospray ionization source.Based on the method described above, the qualitative identifications of the 86 GCs were accomplished without the contrast of standard substances.The results demonstrated that the linear range of this method was from 2 μg/L to 200 μg/L with good correlation coefficients of R2>0.99.The average recoveries of the 86 GCs ranged from 66.2% to 112.8%, and the relative standard deviation (RSDs) was 4.6%-13.9% at three different spiked levels.The limit of detection (LOD) and quantification (LOQ) were 0.006-0.015 mg/kg and 0.02-0.05 mg/kg, respectively.The method is simple, efficient, reliable and accurate, and is suitable for high throughput screening of 86 GCs added illegally in cosmetics.

OBJECTIVE:To investigate the relationship between Tacrolimus blood concentration and effectiveness after renal transplantation and to find factors affecting Tacrolimus blood concentration METHODS:Steady-state trough concentrations of Tacrolimus were collected from 80 renal transplantation patients RESULTS:The effective blood concentration of Tacrolimus was associated with course of treatment CONCLUSIONS:The therapeutic concentration of Tacrolimus was between 12ng/ml and 15ng/ml during the first 2 weeks after renal transplantation,then should be 8ng/ml～15ng/ml

Aim To investigate the effect of isocorydine on arrhythmia in rats induced by myocardial ischemia/reperfusion injury. Methods SD rats were randomly divided into 6 groups: sham group, myocardial ische-mia/reperfusion group, verapamil group and isoc-orydine group of 2. 5 , 5 , 10 mg · kg-1 , each group having 16 rats. Left anterior descending ( LAD ) was tied up to establish the injury model of myocardial is-chemia. ECG was recorded for analysis. The ischemic and infarction area was measured and indexes of SOD, MDA, GSH-Px in the myocardial were determined. Re-sults Isocorydine could significantly reduce the inci-dence of arrhythmia induced by ischemic/reperfusion, including VT, VF;and reduce ischemic and infarction area. Further study demonstrated that isocorydine could increase myocardial SOD and GSH-Px, but de-crease myocardial MDA. Conclusion Isocorydine has a protective effect on the myocardial ischemia/reperfu-sion injury, which might be related to its anti-oxidative function.

Objective To optimize the preparation technology ofCangai volatile oil dextrin inclusion compound/ in situ nasal thermosensitive gel by the central composite design-response surface method.Methods In the design, the investigation factors were the amounts of poloxamer 407 and poloxamer 188, and the evaluation index was the gel temperature. Quadratic models were used to evaluate the mathematic relation between the evaluation index and two investigation indexes to identify the optimum prescription, and then the optimum prescription was verified. Results According to the quadratic models, it was found that there was reliable quantitative relation between the evaluation index and two investigation indexes, among which the optimum dosage was 19.37% for poloxamer 407 and 2.73% for poloxamer 188.Conclusion The optimum model ofCangai volatile oil dextrin inclusion compound/ nasal thermosensitive gel can be obtained from central composite design-response surface method based on quadratic models. This method is reliable and feasible, which can realize the prescription optimization of the in situ gel.

Objective:The aim of this study was to investigate the prognostic value of vitamin B12 as the non-invasive biomarker to predict long-term rebleeding rate in cirrhotic patients with esophagogastric varices.Methods:From Dec 1, 2016 to Dec 31, 2017, cirrhotic patients with esophagogastric varices who had been admitted to Zhongshan Hospital affiliated to Fudan University were enrolled. All these patients received endoscopic treatment to prevent variceal rebleeding. The serum vitamin B12 and folic acid levels were measured in all of them. The receiver operating characteristic (ROC) analysis, Kaplan-Meier analysis, univariate and multivariate cox regression analysis were conducted to explore the value of vitamin B12 in predicting 3-year variceal rebleeding in cirrhotic patients with esophagogastric varices after endoscopic treatment.Results:115 patients were included. The ROC curve analysis indicated that the optimal cutoff value of vitamin B12 for 3-year variceal rebleeding was 567.25 pg/ml. According to the cut-off value, the patients were divided into high-level vitamin B12 group ( n=49) and low-level vitamin B12 group ( n=66). Compared with the low vitamin B12 group, the high vitamin B12 group had lower albumin level, less male (63.3% vs 80.3%), and higher 3-year rebleeding rate ( P<0.05). Cox analysis showed that vitamin B12 and platelet were independent prognostic factors for 3-year rebleeding in patients with variceal bleeding. Conclusions:Elevated peripheral blood vitamin B12 predicts a higher risk of long-term rebleeding in patients with liver cirrhosis and esophagogastric varices.

Objective To evaluate the expression and clinical significance of human papillomavirus(HPV)L1 caspid protein in cytologic specimens of the cervix.Methods The HPV L1 caspid protein in cytologic specimens of the cervix was detected by using immunochemistry in 309 women treated in Shenzhen Hospital of Peking University from Jan.2008 to May 2009.According to histological biopsy,of 309 liquid-based cytologic smears,there were 33 cases of normal cervixes or chronic cervititis,168 cases of grade I of cervical intraepithelial neoplasia(CIN I),84 cases of grade Ⅱ/Ⅲof CIN,and 24 cases of squamous cell carcinomas(SCC).Results The positive expression rate of HPV L1 caspid protein was 27.3%,66.7%,25.0%,and 0 in the normal cervixes or chronic cervititis,CIN I and CIN Ⅱ/Ⅲ,and SCC respectively.There was significant difference between CIN I or SCC and the normal cervixes or chronic cervititis,between CIN I with CIN Ⅱ/Ⅲ,and between CIN I or CIN Ⅱ/m with SCC(all P<0.0 1).The positive expression rate of HPV L1 caspid protein was reduced with the lesion progression from CIN I to CINⅡ/Ⅲ to SCC.309 cases were divided into two groups by 30 and 40 years of age,and there was no significant difference in the HPV L1 caspid protein expression between different age groups(P>0.05).The positive expression rate of HPV L1 caspid protein in the over-1000 RLU/PC viral load group was significantly higher than in the under-1000 RLU/PC viral load group(P<0.05).The sensitivity,specificity,positive predictive value,negative predictive value of HPV L1 caspid protein for detecting high-grade cervical lesions were 80.6%,60.2%,52.1% and 85.2%,respectively.Conclusion The positive rate of HPV L1 caspid protein is decreased with the lesion progression from CIN I to CIN Ⅱ/Ⅲto SCC.HPV L1 caspid protein among HPV-positive women can be a helpful molecular biomarker in risk assessment and prognostic prediction.