OBJECTIVE To explore the predictive value of human telomerase RNA gene component(hTERC) gene amplification and high-risk human papilomavirus(HR-HPV) testing in cervical intraepithelial neoplasia(CIN) as a marker for early diagnosis of cervix carcinoma.METHODS Fluorescence in situ hybridization(FISH) was used to detect the amplification of hTERC of cervical epithelial cells in 72 cases.By using hybrid capture 2(HC-2),two types of the HR-HPV DNA(HPV16/18) of each case were detected.Then,the results were compared with the pathologic diagnosis.The dual-color probe we used was GLP TERC/CSP 3.HeLa cells and lymphocytes from normal marrow were the positive control,while the cervical specimens from healthy outpatients were the negative control.RESULTS hTERC Gene amplification of specimens was tested in 72 cases,the positive amplification rate of hTERC gene in the cervicitis/CINⅠgroup and normal,compared to the cervical carcinomas(100%) and CIN Ⅱ/Ⅲ(68.75%),which showed a significant difference.The rates in CINⅡ and CINⅢ were 60.00% and 83.33%,respectively,which showed a significant difference compared with normal and CINⅠ/inflammation groups.hTERC gene amplification was positive in both HeLa cells and lymphocytes from normal marrow and HC-2 testing was positive in 32 cases of patients containing 11 cases of CINⅡ/Ⅲ,3 cases of cervical cancer,18 cases of cervicitis/CIN1 diagnosed.The positive predictive value(PPV) and specificity(Sp) of hTERC for the high-grade CIN was significantly higher than the PPV and Sp of HC-2 HR-HPV testing.CONCLUSIONS hTERC Gene involves in the progression and occurrence of cervical intraepithelial neoplasia and cervical squamous carcinoma.As a marker for early diagnosis of cervical intraepithelial neoplasia and cervical squamous carcinoma,the FISH method for hTERC gene is more reliable to differentiate the malignant diseases from the benign ones in cervixes than HC-2 HR-HPV DNA testing.The combined detection of HR-HPV and hTERC gene will provide more effective and suitable management to enhance the early diagnosis rate of cervical intraepithelial neoplasia and cervical squamous carcinoma.

Asphyxia ; etiology ; Child ; Dysostoses ; complications ; Female ; Humans ; Osteochondrodysplasias ; Thorax ; abnormalities

Objective To synthesize Wye-125132,an inhibitor of mTOR,and to establish a synthetic route for industrial production.Methods Barbituric acid was used as the raw material to synthesize the intermediate 2, 4, 6-trichrolo-pyrimidine-5-carbaldehyde 2 via Vilsmeier-Haack and chlorination reaction, while intermediate 5 was prepared via 3-step reaction from 1,4-dioxaspiro-[4,5]decan-8-one.The condensation of 2 and 5 was followed by substitution reaction to obtain the key intermediate 7.The side chain 9 was prepared by 3-step reaction from bromine aniline.Then the title product 1 was obtained with the reaction of Suzuki cross-coupling of 7 and 9.The structures of intermediate and target compounds were confirmed by MS and 1 H-NMR.Results and Conclusion Compared with the method reported in the literature,this new synthesis method possesses some advantages, such as ready availablity of raw materials, simple operation, mild reaction conditions and easy disposal of products.The total yield is 13.2%,and the purity of the target compound is 99.77%.

Objective To investigate the properties of the serum free binding of vascular endothelial cells(ECs) to lipopolysaccharide (LPS). Methods The binding of ECs to fluorescein isothiocyanate labelled LPS(FITC LPS) was observed with laser confocal microscope(LCM). The extract of ECs membrane after binding with FITC LPS was recorded with fluorescence microscope. Results The binding of ECs to FITC LPS was in a dose dependent manner. The fluorescence was mainly distributed in cytoplasm, especially near the nucleus which could also be stained. The bound FITC LPS had a very close relationship with the membranous system of ECs. Conclusion ECs could bind to LPS directly and could function intracellularly.

To improve students' experimental research skills,innovative consciousness,promote the construction of functional experimental center,we established series of “comprehensive,contrivable,innovative” experiments in medical students.

Objective Increasing evidence supports the involvement of autophagy in the etiopathology of autoimmune diseases.Systemic lupus erythematosus (SLE) is a potentially fatal autoimmune disease characterized by production of multiple autoantibodies through poorly understood mechanism.In order to explore the role of autophagy in the development of SLE,the expression of autophagy related gene microtubule-associated protein 1 light chain 3 (MAPLC3) in peripheral blood mononuclear cells (PBMCs) was measured in patients with SLE.Methods The mRNA levels of LC3 in PBMCs from 56 SLE patients and 45 healthy individuals were detected by real-time quantitative polymerase chain reaction (qPCR) technique.Autophagy in PBMCs was also determined by flow cytometry (FACs) in 20 SLE patients and 15 healthy controls.The correlation between LC3 mRNA expression and disease activity of SLE (SLEDAI) was then analyzed.Results The mRNA level of LC3 (RQ) in SLE patients was obviously downregulated compared with that in healthy population (1.30 ± 0.10 vs 1.35 ± 0.09; P =0.029),paralleled with the decreased autophagy rate detected by flow cytometry in PBMCs of SLE patients [(2.21 ± 1.07) ％ vs (9.91 ±4.01) ％ ;P =0.047].Moreover,LC3 mRNA expression level was negatively correlated with SLEDAI (r =-0.337,P =0.023).However,when the clinical features of 27 SLE patients with decreased LC3 mRNA expression (RQ ＜ 1.351) were compared with those of other 29 SLE patients with normal or high LC3 mRNA expression (RQ ＞ 1.351),increasing rates of arthritis,serositis,hematological abnormalities were noted in patients with decreased LC3 mRNA expression yet without statistically significance.However,there was a significant difference between two groups in the incidence of renal involvement (P =0.028).Conclusion The impaired autophagy due to dowrnregulated LC3 mRNA level in SLE patients indicates that autophagy plays a role in mediating the occurrence and development of SLE.

Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages.Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor.Statistical tests were performed to analyze the data.Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages.Blood pressure values in the elderly group were higher than those of other groups.The morbidity of hypertension in the elderly group was higher than those of the other groups.Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age.The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups.Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension.Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages.Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.

Objective To explore the clinical value of applying digital technology to make the course of toe vein clear before toe free flap transplantation for thumb reconstruction.Methods Before operation,scan the donor and recipient site by CT,and apply digital three-dimensional CT angiography reconstruction technology to got threedimensional images of the arteries and veins in the donor site for 22 various defected thumbs which were going to have toe free flap transplantation operation,thus indentifyed the types of FDMA and the distribution and course of the toe vein from June,2012 to March,2013.Used computer assisted anatomical modeling to analyze three-dimensional images of blood supply of the donor site and degree of defect of the recipient site and guide the operative design in the donor site according to the digital three-dimensional reconstruction technology.Conducted toe free flap transplantation for thumb reconstruction in accordance with the design.Results It shows that 64-slice dual-source spiral CT could well display the size,distribution and course of toe vein.The toe veins were clarified to two groups,deep one and superficial one whose diameter was larger and was the main reflux vein of the toe.Dorsal vein of the toe origind from venous plexus in the nail bed and it travels along the back of the toe to reach to the near side of metatarsophalangeal joint.And then it beeomes three to four dorsal metatarsal veins to infused dorsal venous arch of foot.The rule of its course was:centralize-decentralize-connect-decentralize again.The great saphenous vein origins from the inner side of dorsal venous arch of foot,rises along the crus before malleolus medialis and collect three larger tributaries:medial malleolar branches-anterior crural branch-outside crural branch.The small saphenous vein derives from the outside of dorsal venous arch of foot and winds behind lateral malleolus up to the back of the curs and infuses to the vein.Conclusion It is useful for venous return,reducing the incidence of vascular crisis by applying toe flap to reconstruct thumbs and clarifying the course and distribution of toe vein with the aid of digital three-dimensional technology to design operation,thus improving the survival rate of toe flap transplantation.

RhoA, a small GTPase, is involved in a wide array of cellular functions in the central nervous system, such as cell motility, cytoskeleton rearrangement, transcriptional regulation, phagocytosis and cell growth. It is not known how spinal cord injury (SCI) affects the expression of RhoA in different nerve cells. In the present study, we investigated the changes of RhoA expression in remote areas of the injury at the 3rd, 7th and 30th day after SCI, which was established by T10 contusion method. Moreover, we examine its expression profile in neurons, astrocytes and microglia. RhoA was found to be weakly expressed in these nerve cells in normal spinal cord. Western blotting showed that, after SCI, the total RhoA expression was up-regulated, and the RhoA expression was increased and peaked at the 7th day. Double immunostaining revealed specific and temporal expression patterns of RhoA in different nerve cells. The expression of RhoA in neurons started to increase at day 3, peaked at day 7 and then decreased slightly at day 30. Expression of RhoA in astrocytes increased moderately after SCI and peaked at day 7. There was no obvious change in RhoA expression in microglia after SCI in remote areas. This study demonstrated that, after SCI, RhoA expression exhibited different patterns with different nerve cells of spinal cord. RhoA expression patterns also changed with time after SCI, and among different nerve cells in the injured spinal cord. These findings can help us better understand the roles of RhoA in SCI.

Objective To verify the methodology for auditing dosimetric parameters in reference and non-reference conditions with thermoluminescent dosimeters (TLDs).Methods Under reference and non-reference conditions,the established TLD methods were used to observe the absorbed dose variations with depth,SSD,field size and 45 wedges for 10 photon beams at 5 hospitals.Dosimetric parameters,including doses at Dmax points in axis,on 5 electron beams of 9 MeV were measured.The measurement results were compared between the TLDs and plane parallel ionization chambers.Results For 6 MV photon beams,the relative deviation of between finger ionization chamber method and TLD chips was in the range of-1.7％ to 5.4％ under on-axis non-reference conditions,and-6.3％ to-0.6％ under off-axis non-reference conditions,respectively,all within the range of ≤ ± 7％ as required by the IAEA.The relative deviation between plane parallel chamber and TLD method was-2.3％ to 3.7％,within ± 5％ as required by the IAEA.Conclusions It is convenient and feasible to use TLD method for quality audits of dosimetric parameters in radiotherapy.