Objective To study the time relationships between apoptosis of neurons and endotheliocytes with the expression of Bcl 2 and Bax after reperfusion of focal cerebral ischemia in rats. Methods Coronal sections of brain were analyzed using an in situ terminal deoxynucleotidyl transferase mediated biotinylated deoxyuridine triphosphate nick end labeling(TUNEL)and immunohistochemical staining methods to observe apoptosis of neurons and endotheliocytes,and expression of Bcl 2 and Bax after reperfusion(2,6,12,24hours and 2,3,7,14,21 days)of focal cerebral ischemia. Results 1.In the ischemic penumbra,apoptotic cells were increased at 2h after reperfusion,peaked at 12 24?h,then decreased successfully for 7 14?days.The time of apoptotic endothelial cells was 12?h later than that of apoptotic neurons.2.The expression of Bcl 2 protein began at 2?h after reperfusion,peaked at 12 24?h,and decreased for 7 14?days.3.Bax protein expressed from 6?h after reperfusion;peaked at 24 48?h,and lasted for 14days.4.The time phase of Bcl 2 expression was similar to the Bax is but later than it. Conclusion\ Apoptosis was a pattern of cell death after cerebral ischemia/reperfusion.The time of apoptotic endothelial cells was later than that of apoptotic neurons.Bcl 2 and Bax play a regulatory role in the apoptotic process.\;[

Organ system based integrated teaching model has been adopted in several medical schools, and these schools face some challenges in this teaching process. In order to provide new sights for organ system based integrated teaching reform in eight-year medical education program, ex-ploration of setting up three-staged-dimensional integrated teaching models was conducted in urinary system by hematuria, and designing related teaching objectives, teaching programs and assessment form to optimize teaching quality in this teaching methods, according to characteristics of the urinary systemic diseases, cultivating objectives of eight-year medical education program and teaching experi-ence of Xiangya School of Medicine.

Objective To explore a new methodology for surgical treatment of severe retracted nipple in women. Methods 20 patients with inverted nipple and 4 recurred patients were involved in this study. Firstly, the site of neonipple tip was marked in the central part of the inverted nipple and its mean diameter usually was 1.2-1.5 cm. Then two shallow and deep triangular pedicled flaps were designed, respectively, in both superior and inferior areas near areola. With temporary traction of the nipple apex provided by a stay suture, the fibrotie bands underneath the nipple base might be cautious-ly released. Moreover, the shallow skin flaps should be about 0.5 cm in thickness and their blood sup-ply was from the subdermal arterial rete of the areola, which were used to cover and reconstruct the neck area of neonipple after a clockwise rotation and advancement simultaneously. While the deep fas-cia tissue flaps were revolved and advanced either horizontally to the opposite pedicle or upward to the inner tip through the tunnel underneath the nipple base in order to improve the height or width of the neonipple neck and prevent flattening as the supporting tissue and their blood supply was from some small perforating branch arteries in the deep part of mammary gland. Finally, purse-string suture was necessary in the base of neonipple which played a key role in avoiding recurrence of nipple inversion. Four vertical diamond-shaped excision-suturation treatment in neck area could make improvemts on the height of those stout and short nipples. Results In all 24 cases corrected by shallow and deep triangu-lar flaps rotation, after 3-6 months' follow-up, there were no complications related to surgery such as infection, hematoma, permanent sensory disturbance, or nipple necrosis, and postoperative recovery was rapid and uneventful. Especially, follow-up data revealed no evidence of recurrence of inversion and all patients were satisfied with their results. Conclusions Triangular flaps and fascia-tissue flaps in shallow and deep areola rotation is effective and easy to be popularized in correction of inverted nip-ple. This technique can improve both the diameter and height of the nipple, and certainly lower the re-currence rate of nipple inversion and achieve good aesthetic results.

Objective To construct the recombinant adenoviros containing heat shock protein70 (Hsp70) gene driven by carcinoembryonic antigen (CEA) promoter. Methods Hsp70 gene and CEA promoter were amplified by RT-PCR and PCR, and then subcloned into the shuttle vector pDC316 to construct the recombinant vector PDC316-pCEA-Hsp70. The recombinant vector was co-transfected with adenoviral backbone plasmid into HEK293 cells to generate the recombinant adenovirus Ad5-pCEA-Hsp70. The recombinant adenovirus was purified by CsCl banding and titrated by 50％ tissue culture infective dose (TCID50) assay. After transfection of the recombinant adenovirus into human pancreatic cell lines SW1990 and BxPC3, the expression of mRNA and protein level of Hsp70 were determined by RT-PCR and ELISA,respectively. Results Digestion and DNA sequencing certified that the Hsp70 gene and CEA promoter was successfully inserted into pDC316 plasmid. Virus acquired through co-transfection with backbone plasmid was confirmed to be constructed successfully by PCR amplification. The particles finally expressed was 2.2 ×1011vp/ml, and the titer was 1.5 x 1010 PFU/ml. BxPC3 cancer cells with positive CEA expression showed increased expression of Hsp70 mRNA and protein after infected by recombinant adenovirus; while SW1990 cancer cells with negative CEA expression showed no change of expression of Hsp70 mRNA and protein after infected by recombinant adenovirus. Conclusions The recombinant adenovirus Ad5-pCEA-Hsp70 which can express Hsp70 gene in CEA positive cancer cells is constructed successfully.

Objective To establish an ideal focal cerebral ischemia reperfusion model in monkeys.Methods Adult healthy rhesus monkeys(Macaca mulatta) 12 cases(male 6 and female 6),aged 4-7 years and weighted 4.8-7.5kg.were used in this study.The middle cerebral artery occlusion/reperfusion(MCAO/R) model was established by inserting a standard micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the proximal segment of the middle cerebral artery(MCA).The regional cerebral blood flow of MCA was occluded by expanding the micro-balloon to cause ischemia,and withdrawing the micro-ballon catheter to reperfuse the MCA.The MCAO/R model was evaluated by angiography,magnetic resonance angiography(MRA),magnetic resonance imaging(MRI),tetrazolium chloride(TTC) staining and neurological behavoral function scores.Results By inserting a micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the MCA,the micro-balloon catheter could be inserted into the MCA to occlude blood flow,and no image of MCA shown on TV screen.In MCA blood flow supplied area,magnetic resonance T1,T2 and DWI showed high signals,TTC staining showed cerebral ischemic infarction,and correspondly the monkeys showed neurological function disorders.This method used a simple operatire procedure had a high successful rate,and could be repeated.Conclusion We showed ideal method to establish the MCAO/R model in monkeys by inserting intraluminally a micro-balloon catheter into the MCA.

Objective To investigate the expression of fibroblast growth factor-2 (FGF-2) and fibroblast growth factor receptor-4 ( FGFR-4 ) in the papillary thyroid carcinomas ( PTC ) and clinical significance . Methods Immunohistochemistry and Western blotting for the expression of FGF-2 and FGFR-4 were performed in 89 cases of PTC and 30 cases of normal thyroid tissues ( NTT) adjacent to the tumors .Results Immunohistochemistry results showed that , FGF-2 and FGFR-4 expressions were high in thyroid carcinoma (P<0.01,P<0.01) in contrast to that in the normal thyroid tissues, and the difference was statistically significant;There was a positive linear correlation between expressions of FGF-2 and FGFR-4 and lymph node metastasis (χ2 =14.798,P<0.01;χ2 =7.27,P<0.01)and differentiation degree (χ2=13.824,P<0.01;χ2 =16.921, P<0.01) in papillary thyroid carcinoma ,while there was no difference in gender ,age and tumor size(P>0.05).Analyzed by Western blotting technique ,FGF-2 and FGFR-4 expressions in thyroid carcinoma were significantly higher than that in normal tissue ,with decrease of cancer degree of tissue differentiation and significantly up regulated expression (P<0.05).Expressions of FGF-2 and FGFR-4 were in a positive linear correlation in the disease (rs=0.434,P<0.01).Conclusion The expressions of FGF-2 and FGFR-4 are correlated with papillary thyroid cancer and they participated in the process of invasion and metastasis , both of which have a positive synergistic effect .The degree of malignancy and biological behavior are meaningful and comprehensive indicators ,which provide a theoretical basis for the subsequent experimental studies of cellular and molecular biology .

Objective To explore the optimal experiment conditions of CCK-8 and MTS for cell proliferation assays in human amniotic epithelial cells and to evaluate the cytotoxicity of these reagents. Methods Human amniotic epithelial cells (hAECs) in logarithm growth stages were prepared in different cell concentrations with DMEM/F12 and 10% FBS. The sensitivity and optimal wavelengths was determined based on the optical density (OD) measured at 450 nm and 492 nm. The optimal time was determined under the conditions of the same cell concentration and defined OD values. HAECs were treated with DMSO, CCK-8 and MTS for 1 h, 2 h, 3 h, and 4 h, respectively. 24 h later, cytotoxicity of the CCK-8 and MTS was evaluated by determination of cell proliferation and Trypan Blue staining. Results The optimal detection wavelength was 450 nm for CCK-8, and 492 nm for MTS. The sensitivity of CCK-8 was slightly lower then that of MTS. The optimal time for incubation hAECs with CCK-8 was 4 h within 1~4 h. The inhibitory on cell proliferation and cytotoxicity of CCK-8 were weaker then those of MTS. Conclusion CCK-8 is a convenient reagent with low cytotoxicity for detection of the proliferation of hAECs.

OBJECTIVE: To summarize the clinical characteristics of testicular torsion and explore the relationship between its incidence and temperature, the value of supplementary examinations, and the factors causing misdiagnosis and testicular lesions. METHODS: We retrospectively analyzed the clinical data of the patients with testicular torsion diagnosed in 9 hospitals in Hunan Province from May 1, 2009 to November 1, 2018. The temperature at onset and WBC value after onset were analyzed. The misdiagnosis rate and testicular lesion rate in the first-visit hospitals were compared between the large general hospitals and local hospitals. The factors that affected the diagnosis and treatment of testicular torsion were analyzed. RESULTS: A total of 274 cases were included in this study, and misdiagnoses occurred in 151 cases (61.9%). The initial diagnosis was made in large general hospitals in 59 cases; misdiagnosis occurred in 30.5% of the cases and the rate of testicular lesions was 57.6%. Of the 215 cases first diagnosed in local hospitals, only 82 cases were diagnosed correctly, and the misdiagnosis rate was as high as 61.9%; the rate of testicular lesion was 85.6% in these cases. There were significant differences in the misdiagnosis rate and testicular lesion rate between the two groups ( < 0.05). CONCLUSIONS The first-visit misdiagnosis rate of testicular torsion is much lower and the probability of testicle preservation is significantly higher in large general hospitals than in the local hospitals, suggesting the importance of health education for testicular torsion, enhancing the awareness of this condition and training of the surgeons in primary hospitals.
Diagnostic Errors ; Humans ; Incidence ; Male ; Retrospective Studies ; Spermatic Cord Torsion ; Testis