Objective To investigate the inhibitory effects and the mechanism of autophagy gene beclin 1 on cervical cancer HeLa cells. Methods The eukaryotic expression vector and short hairpin RNA (shRNA) expression vector of beclin 1 were transfected via lipofectamine into HeLa cells. Experimental cells were classified into 5 groups: pcDNA3. 1 ( + )-beclin 1 group, pSUPER-beclin 1 group, pcDNA3.1 ( + )group, pSUPER group and HeLa group. Real time-PCR and western blot were used for detecting expression of mRNA and protein of beclin 1 and caspase-9 in transfected cells. Flow cytometry was employed to observe the effect of transfection on the apoptosis, and autophagy of HeLa, while proliferation was analyzed by methyl thiazolyl tetrazolium (MTT) assay. The ultrastructural analysis of autophagic vacuoles was under the electron microscope. Five groups cells were seeded subcutaneously on nude mice. The carcinogenic and growth activities of cancer cells in vivo were observed, and immunohistochemistry was used to detect the protein expression of beclin 1 in tumor tissue. Results ( 1 ) The mRNA expression of beclin 1 and caspase-9: pcDNA3. 1 ( + )-beclin 1 group were 994.72 ±468.76 and 12. 88 ±2. 71, pSUPER-beclin 1 group were 0. 18 ± 0. 63 and 0. 11 ± 0. 08, pcDNA3. 1 ( + ) group were 0. 57 ± 0. 12 and 4. 28 ± 3. 25,pSUPER group were 0. 67 ± 0. 29 and 2. 77 ± 1.27, and HeLa group were 0. 74 ± 0. 25 and 3.67 ± 3.78,respectively. The eukaryotic expression vector pcDNA3. 1 ( + ) -beclin 1 significantly improved the expression of mRNA of beclin 1 and caspase-9 in HeLa cells( P ＜0. 05 ), and the shRNA expression vector inhibited the expression of mRNA of beclin 1 and caspase-9 ( P ＜ 0.05 ). (2) The cell proliferations: pcDNA3.1 ( + ) -beclin 1 vector significantly inhibited the growth of HeLa cells, while pSUPER-beclin 1 vector significantly improved the growth of HeLa cells(P ＜0. 05). (3) The rate of apoptosis: pcDNA3.1 ( + )-beclin 1 group was (28.2 ±2.3)%, pcDNA3. 1( + ) group was(14.6 ±4.6)%,pSUPER-beclin 1 group was(5.7 ±2. 0) %, pSUPER group was( 16. 2 ± 3.1 ) %, and HeLa group was( 11.2 ± 3. 0) %. The pcDNA3. 1 ( + ) -beclin 1 vector significantly increased the apoptosis rate, while the pSUPER-beclin 1 vector significantly decreased the apoptosis rate(P ＜0. 05 ). (4)The activity of autophagy: more autophagy cells were identified in pcDNA3.1( + )-beclin 1 group; the rate of autophagy of five group were( 10. 3 ± 1.5)% in pcDNA3. 1 ( + ) -beclin 1 group, ( 3.6 ± 0. 8 ) % in pcDNA3. 1 ( + ) group, ( 1.2 ± 0. 3 ) % in pSUPER-beclin 1 group, (3.2 ± 1.2)% in pSUPER group and (2.2 ± 1. 1)% in HeLa group, there was statistical significances between test groups and control groups( P ＜ . 05 ). (5)Carcinogenic activity of HeLa cells in nude mice: the duration of tumorigenesis was the longest in pcDNA3.1 ( + )-beclin 1 group and the shortest in pSUPER-beclin 1 group among all groups. The tumor size began to grow larger from 7th day after injection in pSUPER-beclin 1 group than in control groups( P ＜ 0. 05 ). The tumor size was smaller from 21st day after injection in pcDNA3.1( + )-beclin 1 group than in control groups(P ＜0. 05). From 28th day after injection,the tumor weigh was (0. 52 ± 0. 08 )g in pSUPER-beclin 1 group, apparently more than HeLa group (0. 37 ±0. 12) g and pSUPER group (0. 34 ± 0. 24 ) g ( P ＜ 0. 05 ). While in pcDNA3. 1 ( + )-beclin 1 group the tumor weighed (0. 18 ±0. 12) g, which was lower than HeLa group and pcDNA3. 1 ( + ) group (0. 34 ± 0. 18 ) g ( P ＜ 0. 05 ) . Conclusions Autophagy gene beclin 1 overexpression can inhibit proliferation and growth of HeLa cells in vitro and in vivo. Beclin 1 not noly participate in the regulation of autophagy signaling, but also play an important role in the regulation of endogenous apoptosis signaling through caspase-9. So it might be one of the new strategies for gene therapy of cervical carcinoma.

Thyroidectomy is an effective treatment for thyroid diseases,especially thyroid malignancies.The postoperative phonological disturbances in patients with recurrent laryngeal nerve injury caused by vocal cord paralysis have been recognized.However,in clinical work,many patients in the absence of recurrent laryngeal nerve injury in the case of postoperative speech changes.With the development of modern electronic acoustics detection technology,the quality of postoperative recurrent laryngeal nerve injury patients with quantitative research,to explore and found that recurrent laryngeal nerve injury after thyroid surgery is not the voice of the factors.

Colorectal neoplasms always present with thickness of the intestinal wall or a soft tissue mass in the enteric cavity. Multi-slice computed tomography (MSCT) with high spatial resolution and advanced post-processing techniques can demonstrate the above signs of the tumor, and the invasive signs of adjacent structures and lymph node metastasis. Combined with three dimensional reformation images, MSCT shows a higher sensitivity than that of double contrast barium enema and electronic colonoscope. MSCT is promising in the diagnosis and preoperative evaluation of colorectal neoplasms.

Less consciousness for science research,lower level of research work,shortage of research team,lack of research institution and so on are the mean problems in clinical hospitals.In recent years,we take the measures of carrying out research training,perfecting research equipment,strengthening the research team,establishing encourage and punish rules.these measures promote the research work of our hospital,change the research sense of hospital staff therefor we get notable progress in research work.

Objective Previously, there was an argument on the surgery way of unilateral papillary thyroid microcarcinoma(PTMC).The purpose of this study was to investigate the predictive factors of occult thyroid carcinoma in unilateral thyroid papillary microcarcinoma, and to compare the surgical methods of unilateral thyroid papillary microcarcinoma.Methods We retrospective analyzed 327 cases of unilateral PTMC patients confirmed by preoperative color Doppler ultrasound examination from January 2014 to January 2016 in Department of Breast Surgery, First Affiliated Hospital of Harbin Medical University.According to the presence of occult cancer, patients were divided into unilateral glandular foci (n=278) and bilateral glandular foci(n=49);according to whether carry out the prevention of central lymph node dissection, patients were divided into lymph node dissection(n=117) and without lymph node dissection(n=210) to further analyze the predictive factors of contralateral gland occult cancer and postoperative complications of preventive central lymph node dissection.Results 55 cases had central lymph node metastasis in 117 cases with lymph node dissection, the metastasis rate was 47.0%.The multiple foci in the ipsilateral gland is a risk factor for the presence of occult foci in the contralateral gland (OR=4.924, 95%CI: 1.605~15.107).There was no significant difference in postoperative complications between lymph node dissection and without lymph node dissection(P>0.05).Conclusion multiple foci in the unilateral thyroid papillary thyroid microcarcinoma can help us to predict the contralateral gland occult cancer, and the central lymph node metastasis rate is higher, therefore, for the unilateral glands of multiple tumors in the papillary thyroid microcarcinoma, we recommended the total thyroidectomy+prophylactic central lymph node dissection surgery way.

In order to enhance the students’knowledge of clinical medicine and lay the foundation for using clinical medicine,we adjusted the content and compiled pharmacognosy teaching materials,aiming at its professional features of clinical pharmacy of Xuzhou Medical College.The pharmacognosy teaching material has been applied in several rounds.Feedback from students suggested that using this teaching material was well-targeted,clear and practical.

Medical research is the core-competitiveness of hospitals.Research management in hospitals with weak scientific research capacity is different from others and has its own demands.People of the management neet to develop themselves to meet the special demands,and can then help develop the hospital.

Objective:To detect anti-LKM-1 antibody with enzyme-linked immunosorbent assay (ELISA) using a recombinant fusion peptide which comprises 257-351 amino acid fragment of CYP2D6 as antigen.Methods:We obtained CYP2D6 cDNA fragment by means of PCR,using total liver cDNA library as the template.The PCR products were ligated into pEGH expressing vector to construct the recombinant expressing vector with high efficiency in Saccharomyces Cerevisiae Y258.The positive clones were identified by PCR reaction and then induced by galactose.Glutathione-Sepharose 4B was used for purification of recombinant CYP2D6 protein.After affinity purification,the antigenicity was identified with Western blot.Serum samples from 26 patients who were positive for anti-LKM-1 antibody,20 patients with other connective tissue disorder(CTD) and 30 normal controls were retrospectively tested with ELISA.Results:A fusion peptide was expressed and purified.The antigenicity was confirmed with Western blot using standard of anti-LKM-1 antibody-positive serum.Of the 26 serum samples which are positive for anti-LKM-1 antibody,5 of 6 samples positive for anti-HCV antibody also recognized the recombinant fusion peptide with ELISA,only one serum sample which was showed positive anti-HCV antibody displayed a negative result in ELISA assay.All other 20 patients with positiv anti-LKM-1 antibody were shown positive in ELISA assay using this recombinant peptide.All the serum samples from patients with other CTD were negative in ELISA assay.Conclusion:The recombinant antigen fragment contains major epitope regions in natural CYP2D6 antigen.Detection of anti-LKM-1 antibody with ELISA using the recombinant peptide can improve the sensitivity and has a potential role in determining its clinical association.

Objective To exlplore the adverse effect of formaldehyde on DNA in isolated mice testicular cells. Methods The testicular cells were exposed to formaldehyde of different concentrations(0, 25, 50, 75, 100 ?mol/L), comet assay was applied to detect DNA breaks. Results Formaldehyde induced DNA breaks at 10 ?mol/L, 25 ?mol/L and 50 ?mol/L (P0.05). The critical concentration at which formaldehyde could induce DNA strand breaks of testicular cells was about 10 ?mol/L, peak point was near 50 ?mol/L. Conclusion Formaldehyde may cause DNA damage in isolated mice testicular cells in various degree.

Objective To study the influence of ion concentration change out of cells on calcium transportation and the relationship between the rising of calcium concentration in the Caco-2 cells and its apoptosis to offer the theoretical and experimental bases for clinical study and digestive tract physiology and patholoogy. Methods Confocal laser scanning microscope(CLSM) and flow cytometry(FCM) were used in this study. Results (1) Diversion of Ca 2+ into cells was increased with the decrease of Fe 3+ concentration out of Caco-2 cells(the final consistence of DFO was 100-300??mol/L),but was hindered with the increase of Fe 3+ concentration out of them(the final consistence fo FAC was 10-100??mol/L) as observed under CLSM; (2) The cell state was fine and its viability was more than 90% as observed under CLSM after treated by A 23187 and Fluo-3/AM(examined with FDA);(3) The Ca 2+ concentration in the Caco-2 cells was increased by A 23187 and this function was dose depended.The Caco-2 cell apoptosis was induced by the increase of Ca 2+ in cells,which was examined with FCM.Conclusion The Ca 2+ transportation was increased with the decrease of Fe 3+ concentration out of Caco-2 cells but was hindered with the increase of Fe 2+ concentration out of them.The Caco-2 cell apoptosis was induced by the increase of Ca 2+ concentration in them.