Abstract:Objective To clone the gene of B-cell lymphoma 6 (Bcl6), construct a green fluorescent expression vector carrying rat Bcl6 gene, and to evaluate its roles in rat liver cells. Methods Total liver RNA was isolated from rat liver samples and cDNA was synthesized by reverse transcription; the coding sequence of Bcl6 gene was obtained by nested PCR. pEGFP-Bcl6 fusion expression vector was constructed by recombination technique and confirmed by sequencing. Then the recombinant vectors were transfected into rat liver cell line BRL-3A by lipofectamine; the expression levels of Bcl6 mRNA and protein were determined by real-time PCR and Western blotting analysis, respectively. Finally, the apoptosis of BRL-3A cells transfected with recombinant vector was analyzed by flow cytometry and cell proliferation was examined by MTT assay. Results The recombinant vector pEGFP-Bcl6 was successfully constructed as determined by PCR and sequencing, and was effectively delivered to BRL-3A cells by transient transfection as confirmed by PCR and Western blotting analysis. Result of flow cytometry proved that Bcl6 up-regulation suppressed apoptosis of rat liver cells and MTT assay found that Bcl6 up-regulation promoted proliferation of liver cells. Conclusion We have successfully cloned rat Bcl6 gene and constructed its green fluorescent expression vector. Our results initially prove that Bcl6 may inhibit cell apoptosis and promote cell proliferation in rat liver cells.

Abstract:Objective To study the effects of umbilical cord-derived mesenchymal stem cells (MSCs) on the growth and metastasis of lung cancer using Lewis lung carcinoma animal models. Methods Animal models of Lewis lung carcinoma were established in 16 C57BL/6 mice and were randomly divided into normal saline (NS) group and MSC group (n=8). Mice in the MSC group were injected with 1×106 MSCs via the tail vein at day 7, 12, and 17. All mice were sacrificed to observe the lung metastases and the tumor size at day 21. Results The average tumor weight was (4.587 5±1.04) g in the NS group and (4.155±1.13) g in the MSC group (P=0.59). The average number of lung metastasis nodules was 3.75±1.39 in the NS group and 1.13±1.13 in the MSC group (P<0.01), showing an inhibitory rate of 70%. Conclusion Umbilical cord-derived MSCs have no effects on the growth of Lewis lung carcinoma in mice, but they can inhibit tumor metastasis.

Abstract:Objective To investigate the radioprotective effects of genistein (GEN) against radiation-induced DNA damage in human liver cell line L-02．Methods （1）L-02 cells were treated with different concentrations of GEN (1, 5, 10, and 20 μmol/L) for 24 h, and then irradiated with X-ray at the doses of 6, 8, 12,16,and 20 Gy. Forty-eight hours after irradiation, MTT method was applied to examine the proliferation of L-02 cells.（2）L-02 cells were treated with different concentrations of GEN (1, 5, 10, and 20 μmol/L) for 24 h, and then irradiated with X-ray at the doses of 8 Gy (300 cGy/min) . Single cell gel electrophoresis was used to determine the DNA damage after radiation . Results (1)After irradiation with 6, 8 and 12 Gy of X-ray, the cell proliferation rate of 5 μmol/L GEN-pretreated group was significantly increased compared to radiation alone (R) group (P<0.05). But no significant increase was observed in GEN-pretreated groups irradiated with 16 Gy and 20 Gy of X-ray compared with R group. (2)As for the DNA damage, no comet cells were detected in normal control group or all GEN-treated groups without irradiation. After irradiation with 8 Gy of X-ray for 24 h, comet incidences were less than 1% in all GNE-pretreated groups and R group, and comet tail length showed no significant difference between different groups. At 48 h after irradiation, the comet incidence of R group was (24.2±1.2)% and the comet tail length was (283.6±22.3) μm, while both comet incidence and tail length of GEN-pretreated groups were significantly lower than those of R group (P<0.05). The comet incidence and tail length of R group were significantly decreased 72 h after irradiation compared with 48 h after irradiation (P<0.05), and those in 1 μmol/L and 5 μmol/L GEN-pretreated groups were still significantly lower than those of R group (P<0.05), but the comet incidence and tail length of 10 μmol/L and 20 μmol/L GEN-pretreated groups were significantly increased compared to those of R group (P<0.05). Conclusion Low concentration of GEN (1 and 5 μmol/L) can effectively protect the human liver cells L-02 against X-ray (8 Gy)-induced DNA damage.

Abstract:Objective To investigate the influence of thyroid hormone T3 preconditioning on interleukin-10 (IL-10) and interleukin-1 receptor antagonist (IL-1Ra) modulation and neutrophil infiltration after renal shchemia/reperfusion (IR) in mice, so as to study the protective effect of T3 on IR kidney. Methods Totally 120 male C57BL/6 mice were randomly divided into four groups (n=30), namely, control group (sham operation), IR group (only received renal IR), T3+IR group (T3 preconditioning for 48 h before renal IR), and NaOH+IR group (received equivalent 0.1 mol/L NaOH soltuion 48 h before renal IR). The serum creatinine and blood urea nitrogen (BUN) were determined 24 h after reperfusion in each group; renal histological damages were scored using PAS staining; the levels of neutrophil infiltration were evaluated by MPO staining, and IL-10, IL-1Ra mRNA expression was examined by real-time PCR at 1, 3, 6, 12, 24, and 48 h after reperfusion. Results The serum creatinine and BUN levels of T3+IR group were significantly lower than those of IR group 24 h after reperfusion (P<0.05), which was accompanied by lower histological score and significantly less neutrophil infiltration (P<0.05). Real-time PCR results showed that IL-10 and IL-1Ra mRNA expression in T3+IR group was significantly higher than that in the IR group (P<0.05) 12 h after reperfusion, which lasted for 48 h after reperfusion. The above parameters were similar between IR group and NaOH+IR group. onclusion Thyroid hormone T3 preconditioning can alleviate renal IR injury, partly by increasing expression of IL-10 and IL-1Ra and subsequently reducing neutrophil infiltration at the late phase of renal IR.

Abstract:Objective To evaluate the prevalence of advanced colorectal neoplasm (ACN) in asymptomatic average-risk Chinese Han population, so as to provide a scientific basis for determining the optimal age for colorectal cancer screening. Methods A prospective cross-sectional design was used in this study. We used a self-made questionnaire to survey the average-risk adults receiving colonoscopy; the subjects were from 19 nationwide representative hospitals. The survey items included the demographic characteristics, life style, dietary habits, colonoscopy examination Results, etc. The risk of ACN was evaluated using previously established average-risk evaluation system, which was composed of eight variables: age, sex, smoking, diabetes mellitus, green vegetables, pickled food, fried food and white meat, with a total score of 0 to 14 points. We calculated the ACN prevalence of each risk level. The χ2-test was used to compare the prevalence rates of ACN between different risk levels, age groups and genders.Results A total of 7,541 subjects received complete colonoscopies. The ACN prevalence rose greatly with the increase of risk score. In the population with a risk score of 0-2, the ACN prevalence was 1.3%-4.0% in those younger than 70 years and 8.9%-15.3% in those elder than 70 years(P<0.01). In the population with a risk score of 3-4, the ACN prevalence was 3.6% in those younger than 45 years old and 5.4%-14.8% in those elder than 45 years (P<0.01). In the population with a risk score higher than 4, the ACN prevalence was at a high level (9.8%-22.7%), with the prevalence being 9.8% in the 40-44 years group, which was higher than those in subjects of 70-74 years old with a risk score of 0-2(8.9%). The overall ACN prevalence was 2.9%-4.6% in subjects younger than 55 years old and 6.8%-17.6% in those elder than 55 years (P<0.01). The high-risk age of ACN was elder than 45 in male and elder than 60 in female. Conclusion The optimal starting ages for colorectal cancer screening are different for subjects with different risks and genders. The starting ages for those with risks scores of 0-2, 3-4, and >4 were 70 years, 45 years and 40 years, respectively. The optimal starting age for screening in women can be 15 years later than in men.

Abstract:Objective To investigate the attitude on pre-exposure prophylaxis (PrEP) among drug users from high-risk population of AIDS in western China and its influencing factors. Methods A total of 190 drug users were recruited by snowball sampling from high-risk population of AIDS including those involved in mem having sex with men (MSM), female sex workers (FSW) and the spouse or sex partner (PAR) of HIV carrier in Chongqing, Sichuan, Guangxi and Xinjiang. Self-administered questionnaire survey was conducted with the assistance of investigators. Univariate and multivariate logistic regression was employed for statistical analysis. Results MSM, FSW and PAR accounted for 34.74%(66/190), 48.42%(92/190) and 16.84% (32/190) among the 190 drug users, respectively. The positive attitude rate for PrEP among drug users reached 70% in the premise of drug safety and effectiveness, which increased with favorable condition provided. The Results of multivariate logistic regression analysis indicated that the factors significantly associated with the positive attitude for PrEP included awareness of AIDS seriousness (OR: 2.66, 95% CI: 1.14-6.25, P=0.024 2), attitudes towards HIV patients(OR: 4.41, 95% CI: 1.68-11.58, P=0.002 6； OR: 2.99, 95% CI: 1.05-8.54, P=0.040 3) and virus detection of AIDS (OR: 1.94, 95% CI: 0.98-3.87, P=0.058 1). Conclusion The attitude for PrEP among drug users from AIDS high-risk population is mainly related to the attitude for AIDS, AIDS-related knowledge and behavior, and preventive measures for AIDS, indicating that PrEP should be implemented and promoted with a sound social background, and education on HIV/AIDS prevention should be reinforced. Positive attitude towards AIDS prevention need to be developed among drug users by various behavioral therapies, so as to improve the attitude for PrEP among drug users with high HIV risks.

Abstract:Objective To study the effect of triptolide (TP) on the proliferation of breast carcinoma cell line MCF-7 and its association with P53/P73 gene expression and methylation. Methods MCF-7 cells were treated with different concentrations of TP (10 ng/ml, 20 ng/ml, and 40 ng/ml), and the proliferation of MCF-7 cells was measured by MTT method. The expressions of methyltransferase DNMT1, DNMT3a and DNMT3b mRNA were measured by RT-PCR in MCF-7 cells, P53/P73 gene methylation was analyzed by methylation specific PCR, and the protein expression of p53/P73 in MCF-7 cells was examined by Western blotting assay. Results TP inhibited the proliferation of MCF-7 cells in a dose-dependent manner (P<0.05, P<0.01), with the inhibitory rate being (70.1±3.52)% at 40 ng/ml TP, and the IC50 of TP was 20 ng/ml. TP significantly inhibited DNMT1, DNMT3a, and DNMT3b mRNA expression in MCF-7 cells (P<0.05, P<0.01), and it also significantly inhibited methylation of P53 promoter region. TP increased P53 gene expression at 20 ng/ml and the increase was significant at 40 ng/ml (P<0.01). TP reversed the hypermethylation of P73 gene in MCF-7 cells; it also significantly increased P73 mRNA expression at 10 ng/ml (P<0.05), and the increase was in a dose-dependent anner. Western blotting analysis showed that TP (20 ng/ml) increased the protein expression of P53 and P73 in MCF-7 cells. Conclusion TP can promote the expression of P53 and P73 genes through inhibiting methyltransferase-dependent gene methylation, and further inhibit the proliferation of MCF-7 cells.

Abstract:Objective To investigate the protective effect of Jinlida granules on islet β cells in diabetes mellitus rats. Methods Diabetic models were induced by high-fat diet and intraperitoneal injection of streptozotocin (STZ ) in SD rats. The study was divided into model group, Jinlida granule group (0.75, 1.5, 3.0 g/kg), α-lipoic acid group, insulin group, and metformin group. Each group was given corresponding drugs, with insulin given by subcutaneous injection and other drugs given by intragastric administration. The body weight, fasting blood glucose(FBG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), the activities of superoxide dismutase (SOD) and glutathione(GSH), and the contents of malondialdehyde(MDA), IL-1β and TNF-α were examined 2 months later. The pancreatic tissues were immunostained for insulin. Results Compared with the model group, the levels of FBG and HbA1c were decreased in other groups(P<0.05, P<0.01). The activities of SOD and GSH were significantly increased (P<0.01) and the levels of MDA, IL-1β and TNF-α were significantly decreased (P<0.01) in high-dose (3.0 g/kg) Jinlida granule group, with increased islet area as shown by pathological staining. Conclusion Jinlida granule has protective effect against STZ-induced islet β cell injury in diabetic rats.

Abstract:Objective To evaluate the clinical value of postoperative adjuvant transcatheter arterial chemoembolization (TACE) on the prognosis of hepatocellular carcinoma (HCC) patients after radical hepatectomy using a prospective cohort study. Methods A total of 220 HCC patients underwent radical hepatectomy from Jan. 2008 to Dec. 2008 and 104 were recruited in the present study. Fifty-sixpatients (TACE group) received adjuvant TACE one month after hepatectomy, and 48 (control group) did not receive any adjuvant therapies. Follow-up was done regularly. The disease-free survival (DFS) time and total survival (TS) were statistically analyzed. Results The median DFS time in TACE group was significantly shorter than that in the control group (11[2-38] months vs 13 [2-59] months, P=0.005). The 1-year, 2-year and 3-year recurrence rates were 50%, 85.7%, and 89.3% in TACE group and 46.8%,58.3%, and 62.5% in the control group(P=0.005), respectively. Multivariate COX regression analysis indicated that TACE, AFP, intact tumor peplos, liver cirrhosis, vascular invasion and tumor Edmondson-Steiner grade were the risk factors for recurrence (P<0.05). The median TS time periods in TACE group and control group were 29 (4-41) months and 24 (5-59) months (P=0.789),respectively. The 1-year, 2-year and 3-year survival rates were 85.6%, 59.5%, and 36.5% in TACE group and 75%, 50%, and 41.7% in the control group (P=0.789), respectively. Multivariate COX regression analysis indicated that AFP, intact tumor peplos and tumor Edmondson-Steiner grade were correlated with the total survival of patients (P<0.05). Conclusion Adjuvant TACE can not improve DFS and TS of HCC patients after radical hepatectomy, and it may even contribute to a poor prognosis. Therefore more attention should be paid in choosing treatment strategy.

Abstract:Objective To investigate the effect of radiofrequency catheter ablation (RFCA) on the function and structure of left ventricle during treatment for frequent premature ventricular beats (PVBs). Methods Totally 56 patients with PVBs, who had been successfully treated with RFCA from Jan. 2006 to Dec. 2010, were included in the present study (ablation group). The other 42 patients with frequent PVBs, who received no RFCA treatment, served as controls (non-ablation group). The changes of total number of PVBs, left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), interventricular septal diameter (IVSd), left ventricular posterior wall diameter (LVPWd) were observed before and after different treatments. Results (1)Compared with the non-ablation group, the ablation group had significantly reduced total number of PVBs ([125±113]/24 h vs [17 813±1 542]/24 h, P<0.01), increased LVEF ([0.72±0.06] vs [0.64±0.08], P<0.05), and significantly decreased LVEDD ([44.1±4.0] mm vs [48.9±3.8] mm), LVESD ([27.0±3.7] mm vs [30.1±4.2] mm), IVSd ([7.5±0.8] mm vs [8.5±0.8] mm), and LVPWd ([7.5±0.8] mm vs [8.5±0.9] mm) (P<0.05).(2)Compared with the baseline levels, the total number of PVBs in the ablation group decreased from (23 662±12 559)/24 h before RFCA to (125±113)/24 h(P<0.01) after RFCA,and patients had relieved chest tightness and heart palpitations. Meanwhile, the LVEF was significantly increased after RFCA ([0.72±0.06] vs [0.64±0.09], P<0.05); LVEDD ([44.1±4.0] mm vs [50.6±5.4] mm), LVESD ([27.0±3.7] mm vs [32.1±5.3] mm), IVSd ([7.5±0.8] mm vs [8.8±1.1] mm), and LVPWd ([7.5±0.8] mm vs [8.7±1.1] mm) were significantly decreased after RCFA (P<0.05). Conclusion RFCA can effectively improve the structural remodeling and cardiac dysfunction induced by PVBs, relieving the patient symptoms.

Abstract:Objective To discuss the diagnosis and surgical treatment of symptomatic Rathke cleft cysts (RCCs). Methods Twenty-three symptomatic RCCs, who were verified by microsurgeries and pathological examination from Jan. 2001 to Oct. 2010, were retrospectively analyzed. Nine of them were males and fourteen of them were females. The main clinical manifestations included headache (65.2%), visual disturbance (34.8%) and pituitary dysfunctions (47.8%). The magnetic resonance imaging (MRI) showed 10 pure intrasellar lesions, 10 lesions of intrasellar with a suprasellar extension, and 3 pure suprasellar lesions. Sixteen cases were treated by transsphenoidal microsurgery and 7 were treated by craniotomy. The contents of the cysts were drained totally during the operation, and part of or the total cyst wall were safely moved and kept open. Results Postoperative follow-up period ranged from 14 months to 10 years. All the 8 patients were released from visual disturbance. Headache was improved in 13 of the 15 patients and pituitary dysfunction was improved in 7 of the 11 patients. Recurrence was noted in 1 patient. Conclusion Preoperative definite diagnosis for the symptomatic RCCs is difficult due to a variety of clinical manifestations and MRI signals. Most of the lesions can be treated via transsphenoidal approach. Transcranial approach is necessary for the pure suprasellar RCCs. Microsurgery is an effective strategy for the treatment of symptomatic RCCs, because it can safely relieve the symptoms and make definite diagnosis.

Abstract:Objective To compare the values of cone beam CT (CBCT) with panoramic radiograph in preoperative planning for extraction of mandibular impacted wisdom teeth. Methods Totally 97 patients with 135 wisdom teeth were assessed with panoramic radiograph, and 41 cases presenting 52 impacted wisdom teeth underwent additional CBCT scan were enrolled in this study. Preoperative planning was made for removal of bone, decapitating the crown, sectioning the root and direction of tooth luxation according to the information provided by panoramic radiograph and CBCT images. The Results were processed statistically with SAS 9.3.1 software package for χ2 test. Results Compared with the panoramic radiograph, CBCT provided a more precise and quantitative guide for bone removal, root sectioning and direction of tooth luxation (P＜0.001). The two Methods showed no significant difference in making decision of crown decapitating. Conclusion CBCT can provide comprehensive three-dimensional information during preoperative planning for the extraction of mandibular impacted wisdom teeth. An appropriate surgical approach, with correct tactics and Methods may facilitate the operation, prevent impairment to adjacent structures and reduce surgical complications.

Abstract:Objective To analyze the advantages and disadvantages of two surgical Methods for treatment of lung cancer complicated with coronary heart diseases (CHD), so as to provide evidence for clinical practice. Methods Nine patients with lung cancer complicated with CHD received radical resection of lung cancer shortly after percutaneous coronary intervention (PCI) with stent implantation from Dec. 2010 to Dec. 2011 in the General Hospital of PLA. During the same period eight patients with lung cancer and severe coronary artery stenosis underwent off-pump coronary artery bypass grafting (OPCABG) and radical operation for lung cancer in the Third Hospital of Peking University. The clinical data of the two groups (PCI group and OPCABG group) were analyzed and compared. Results There were no perioperative myocardial infarction, pulmonary infection, incision infection, reoperation due to bleeding, or perioperative death in both groups. Compared with the PCI group, OPCABG group had significantly longer overall operation time ([428±22] min vs [149±32] min), more blood loss ([367±19] ml vs [171±19] ml), and more chest drainage in the first postoperative 24 hours ([527±17] ml vs [250±14] ml, P<0.01). The operation time for lobectomy ([158±27] min vs [149±32] min) and postoperative hospital stay ([10±0.5] d vs [11±0.5] d) were not statistically different between the two groups. Conclusion Coronary intervention and coronary artery bypass grafting have their own advantages in treating lung cancer complicated with severe ischemic heart disease. Appropriate revascularization techniques should be chosen according to indications before radical operation for lung cancer, while considering the patients’ physical condition and specific site of coronary lesions.

Abstract:Objective To investigate the clinical effect of low-dose cyclophosphamide plus prednisone (CP regimen) in treatment of multiple myeloma (MM) and its influence on serum levels of vascular endothelial growth factor (VEGF) and platelet-derived growth factor BB (PDGF-BB). Methods Totally 54 patients with refractory or relapse MM were continuously treated with CP regimen: oral cyclophosphamide (CTX, 50 mg/d) and prednisone (Pred, 15 mg/d). The peripheral blood samples were collected from each group 0, 2, 4, and 6 months after treatment. Serum VEGF and PDGF-BB levels were analyzed by ELISA. Results The overall effective rate was up to 69%(36/52) in our study. The treatment was effective in 30 patients, including 2 complete response (CR) cases, 4 very good partial response (VGPR) cases and 24 partial response (PR) cases, and their serum samples were examined for 4 times.The serum levels of VEGF and PDGF-BB were significantly decreased in the 30 patients at 2, 4, and 6 months after treatment (P<0.01); while their levels were not significantly change in the 7 invalid patients 2 months after treatment compared with before treatment. Conclusion Low-dose cyclophosphamide plus prednisone has noticeable effect in treatment of MM patients; it can also greatly down-regulate serum VEGF and PDGF-BB levels. The mechanisms of CP regimen in MM patients may be associated with the inhibition of new blood vessel generation.

Abstract:Objective To establish a regression equation for drug revenues by analyzing the factors influencing drug revenues in seven military hospitals from 2007 to 2010. Methods The data were obtained from the financial and performance-effect data in 7 hospitals from 2007 to 2010; the database was created by Excel 2003 software. Principal component regression analysis was performed for variables including the hospital scale, amount of service, local economic level, drug add rate using SPSS 18.0 software. Results and Conclusion (1) Hospital scale and amount of service were the critical influencing factors of drug revenues in military hospitals; (2) Per capita cost of outpatient and inpatient also contributed to drug revenues; and (3) drug add rate and local economic level were negative influencing factors of drug revenues.

Abstract:Objective To observe the follicle growth of vitrified rabbit ovarian tissues after autologous implantation in mesometrium in rabbits. Methods Fifteen female New Zealand rabbits were randomly divided into two groups.(1)Vitrified ovarian tissue group (n=10): rabbits were bilaterally ovariectomized and the ovary was made into small tissues for vitrified freezing; two weeks later the frozen-thawed ovarian tissues were autologously transplanted into the mesometrium. (2)Control group included 5 animals. The long-term survival of ovarian tissues, survival and development of follicles, and endocrine function after gonadotropin treatment were evaluated by light microscope and vaginal exfoliocytology. The response of cryopreserved ovarian tissues to gonadotrophin was observed after autologous implantation. The oocytes of MⅡ stage were subjected to intracytoplasmic sperm injection(ICSI)to further observe the development potency of oocytes after autologous implantation. Results The ratios of follicles of each stage 2 months and 6 months after transplantation were similar to those of control group (P＞0.05). Six months after follicle-stimulating hormone (FSH) stimulation, the ratio of mature follicles in ovarian tissues was significantly higher than that in the group untreated with FSH (P＜0.05). The oocytes derived from ovarian tissues after superovulation could obtain normal embryos by ICSI. Conclusion Vitrified rabbit ovarian tissues can achieve ovarian function after autologous implantation in mesometrium, and the function can maintain for a long period; furthermore, normal embryos can be obtained by ICSI technique.

Abstract:Hepatitis C virus (HCV) cell entry is a multistep process mediated by various receptors. Among these receptors, the scavenger receptor class B type Ⅰ (SR-BⅠ) is the one considered to be the first to interact with HCV. SR-BⅠ can bind to HCV envelope glycoprotein E2, in which the hypervariable region 1 (HVR1) segment locating at the N-terminus of E2 protein plays a critical role. The interaction of SR-BⅠ with HCV can not only mediate HCV cell entry, but also attenuate neutralization activity of antibodies against E2 protein, contributing to the HCV immune evasion. Therefore, studying the mechanism of HCV/SR-BⅠinteraction during cell entry can help to identify important targets in the initial step of viral infection, contributing to prevention and treatment of HCV infection. This paper reviews the current knowledge on the biological characteristics of SR-BⅠ and mechanisms of HCV cell entry mediated by virus/SR-BⅠinteraction.

Abstract:Traumatic optic neuropathy (TON) is caused by blunt trauma and has poor prognosis due to lack of effective treatments, and it is a main reason for high rate of disability after traumatic brain injury. Progression in neurobiology and molecular biology has made it a research focus to study optic nerve damage and regeneration. This article reviews the advantages and disadvantages of various animal TON models, paving a way for establishing ideal TON models and for future prospective researches.

Abstract:The continuous expanding of targets is a distinct character of systematic evolution of ligands by exponential enrichment (SELEX) technique and other random library technologys (such as phage display technology). Over the past 20 years the targets of SELEX have expanded to various metal ions, organic molecules, drugs, proteins and biomacromolecules, pathogenic bacteria, live cells and tissue slide. The SELEX screenings using live cells and tissue slide as targets have been preliminarily applied for studying targeted diagnosis and treatment of cancers. This paper reviews the expansion of the targets of SELEX technique.

Abstract:Objective To investigate the expression of endothelin-1 (ET-1), ET-2, ET-3,endothelin receptor A (ETRA), ETRB and endothelin coverting enzyme (ECE) in the retinas of 8-week-old diabetic SD rats. Methods The retinal gene expression profiles of healthy and 8-week-old diabetic rats were constructed with restriction fragment differential display polymerase chained reaction (RFDD-PCR), and the differential expression of ET-1, ET-2, ET-3, ETRA, ETRB and ECE was verified using semi-quantitative RT-PCR and Western blotting analysis.Results The Results of RFDD-PCR showed that the expression of ET-1, ET-2, ET-3, ETRA, ETRB and ECE was up-regulated in diabetic retina. The Results of semi-quantitative RT-PCR and Western blotting analysis showed that the expression levels of the six genes and proteins (relative D ratio) in diabetic group were significantly higher than those in the normal retinas (P<0.05 and P<0.01, respectively). Conclusion The expression of ET-1, ET-2, ET-3, ETRA, ETRB and ECE is up-regulated in diabetic retina, suggesting that the six genes may be involved in the pathgenesis of diabetic retina.

Abstract:Objective To investigate the distribution of HBV genotypes and the mutations in basic core gene promoter (BCP), pre C/C region in clustering family patients infected with chronic hepatitis B (CHB), and to discuss the related clinical significance. Methods A total of 98 CHB patients from 38 family with family clustering features were included in the experimental group, and 110 CHB patients without family clustering features were taken as controls. HBV genotypes were detected in CHB patients by nested PCR with genotype-specific primers. The mutations in BCP and pre C/C region were detected using PCR. Serum ALT, TBIL, ALB, HBV-DNA levels, and hepatitis B antigen and antibody were all examined. Results HBV genotype C was the predominant genotype (52.9%, 36/68) in group of parents and their children, and genotype B was the predominant genotype in group of other infected family members (73.3%, 22/30) and control group (67.3%, 74/110); the frequency of HBV genotype C in group of parents and their children was significantly higher than other infected family members and control group (P<0.01). There were only a few B+C and B+D mixed types. On the whole, HBV genotypes from the same family were basically the same. The frequency of BCP A1762T/G1764A mutations in clustering family CHB patients with genotype C (61.9%, 26/42) was significantly higher than that in CHB patients with genotype C without family clustering features (35.7%, 10/28) (P<0.05), so was the HBV-DNA level (P＜0.01). Conclusion HBV genotype in clustering family is probably associated with the transmission route of virus. HBV genotype C may be easier to transmit from parents to their children. Genotype C and BCP A1762T/G1764A mutations may be associated with the development of CHB in clustering family.

Abstract:Objective To investigate the mutation of low-density lipoprotein receptor (LDL-R) gene in 3 patients with familial hypercholesterolemia (FH) in two generations, so as to discuss the pathogenesis of FH. Methods A 17-year-old patients was selected to undergo physical examination, lipid level test, electrocardiography, cardiac ultrasound and coronary artery angiography. Pedigree analysis was carried out based on family investigation. The promoter and the 18 exons of the LDL-R gene and the flank sequence were amplified by PCR; DNA sequencing was used to detect point mutation. Ninety normal subjects from the native place of the proband and 190 subjects from random population were taken as controls. Results Totally 28 members of 4 generations were examined. The proband, her elder sister and grandaunt had FH with xanthoma, and their total cholesterol (TC) levels were 18.89 mmol/L, 15.23 mmol/L, and 12.89 mmol/L, respectively. Pedigree analysis showed that the genetic pattern of this family was consistent to autosomal dominant inheritance trait. DNA sequencing demonstrated that a G1448A substitution caused a nonsense mutation TGG to TAG in exon 10 of LDL-R gene, a Trp→462 stop mutation. The mutation of the proband, her older sister and grandaunt were homozygous, heterozygous and heterozygous, respectively. The same mutation was not detected in the family members from the proband’s father and people from control group. Conclusion The proband, her elder sister and grandaunt have the same mutation, the Trp→462 stop mutation in exon 10 of LDL-R gene, which might be the key mutation that causes FH in this pedigree.

Abstract:Objective To investigate the expression of claudin-3 and β-catenin in borderline ovarian tumors (BOTs) and its clinicopathological significance. Methods We examined the expressions of claudin-3 and β-catenin and their relationship with the clinicopathological features in 17 normal ovarian tissues, 31 epithelial benign ovarian tumors, 39 epithelial malignant ovarian tumors and 69 BOTs by SP immunohistochemistry method. Results The positive expression rate of claudin-3 protein in BOTs (59.4%, 41/69) was significantly lower than that in the malignant ovarian tumors (89.7%, 35/39), and higher than that in normal ovaries (5.9%, 1/17) and benign ovarian tumors (22.6%, 7/31)(P<0.01); the expressions were not significantly different between the normal ovaries and benign ovarian tumors (P>0.05). The positive expression rate of β-catenin protein in BOTs (52.2%, 36/39) was significantly lower than that in the malignant ovarian tumors (84.7%, 33/39), and higher than that in normal ovaries (11.8%, 2/17) and benign ovarian tumors (29.0%, 9/31; P<0.01 or P<0.05); the expressions were not significantly different between the normal ovaries and benign ovarian tumors (P>0.05).The expression of claudin-3 and β-catenin was correlated with the clinciopathological stage and intraperitoneal metastases (P<0.05), but not with the histological type or age of BOTs patients (P>0.05). Expression of claudin-3 and β-catenin was positively correlated in BOTs (r=0.439, P=0.000). Conclusion Claudin-3 and β-catenin may play a synergistic role in the oncogenesis and progression of BOTs. Simutaneous detection of claudin-3 and β-catenin is valuable for diagnosis and treatment of BOTs.

Abstract:Objective To introduce the clinical experience and effects of retroperitoneal laparoscopic partial nephrectomy in treatment of patients with T1b renal cell carcinoma. Methods Fifteen patients (10 males and 5 females) with T1b renal cell carcinoma who were treated from Nov. 2007 to Jun. 2011 were retrospectively analyzed. The patients had a mean age of (56.2±9.8) years old and a mean tumor size of (5.5±1.4) cm in diameter, with 8 cases on the left side and 7 cases on the right side. All of the 15 patients received retroperitoneal laparoscopic partial nephrectomy. Results All the 15 cases were operated successfully without conversion to open surgery, and there were no severe perioperative complications. The mean operation time was (100±23.6) min, mean warm ischemia time (WIT) was (21.3±8.6) min, and the mean blood loss was (35.3±11.6) ml. Postoperative urinary leakage was found in one case and serum creatinine transient increase was found in one case. The mean postoperative hospital stay was (5.7±1.8) d. All patients had normal renal function and had no tumor recurrence or metastasis during a mean follow-up of (21.2±10.1) months. Conclusion Retroperitoneal laparoscopic partial nephrectomy is safe and effective for treatment of patients with T1b renal cell carcinoma, with less trauma and faster recovery; however, the long-term effects still need large sample studies and long-term follow-up.

Abstract:鼻咽癌是中国南方地区的高发肿瘤,是目前放疗有效率较高的肿瘤之一,但复发和转移仍是导致鼻咽癌患者治疗失败的主要原因。如何完善鼻咽癌的综合治疗,提高其总体生存率是目前鼻咽癌治疗的重点。细胞因子诱导的杀伤(cytokine-induced killer,CIK)细胞过继免疫疗法已经成为肿瘤患者放、化疗后辅助治疗的重要手段之一,对于促进患者免疫系统的重建、消除残留病灶及骨髓保护等都有良好的效果[1-2]。本研究观察了鼻咽癌患者自体CIK细胞的体外增殖能力及杀瘤活性,并与CD3AK和LAK细胞进行比较,旨在为鼻咽癌自体CIK细胞治疗应用于临床提供实验依据。

Abstract:心血管系统疾病是导致终末期肾病和肾移植患者死亡的一个重要原因[1],血管内皮细胞功能损伤是动脉硬化的早期改变,应用高频超声探测肱动脉内皮依赖性舒张与非内皮依赖性舒张,通过对比可无创性地判断血管内皮细胞功能[2]。研究发现尿毒症患者和同种异体肾移植患者的动脉内皮细胞功能均有损伤[3-5],但这些研究的受试者大多伴有高血压、高脂血症、糖尿病等已知心血管危险因素,这些危险因素本身也可以引起动脉内皮细胞功能损伤。本研究排除了所有伴有这些危险因素的受试者,对血透患者和肾移植患者的动脉内皮细胞功能进行比较。

Abstract:1 病例资料患者女性,49岁,以“口齿不清2 d”为主诉于2011年10月27日收入我院神经内科。患者于2011年10月25日晨起出现口齿不清、舌头僵硬,无法发“斯”等平舌音,无发热,无头痛、头晕、恶心、呕吐,无视物旋转、复视、耳鸣,无语言理解障碍,无吞咽困难、饮水呛咳,无肢体麻木、肢体活动障碍,无意识障碍、抽搐、二便失禁。当日16:00症状稍减轻,18:00症状基本缓解。2011年10月26日晨再次出现类似症状。门诊头颅CT检查未见明显异常,为进一步诊治以“构音障碍待查”收入院。发病以来,患者神志清楚,精神、进食、二便正常。既往有“心绞痛”病史20余年,间断服药。无高血压、糖尿病、手术外伤史。自诉2011年发现“甲状腺肿块”,未正规诊治。体检:T 36℃,P 74次/min,R 18次/min,BP 130/90 mmHg(1 mmHg=0.133 kPa)；神清,言语略含糊,口齿欠流利,查体合作,对答切题,定位定向力正常,近记忆力减退,计算力正常。颅神经检查未见异常。运动系统:未见四肢肌萎缩,四肢肌张力正常,肌力Ⅴ级,未见静止性震颤。感觉系统:四肢深浅感觉对称。生理反射存在,病理反射未引出。共济运动正常。脑膜刺激征:颈软,Brudzinski征(-),Kernig征(-)。GCS评分15分。内科体检:甲状腺未及明显肿大及肿块,心肺听诊无异常,眼睑及双下肢无水肿。入院后实验室检查:血、尿、粪常规正常,血糖、肝肾功能、电解质、自身免疫抗体、肌钙蛋白、肌红蛋白、肌酸磷酸激酶、凝血功能正常,抗链球菌溶血素O、类风湿因子阴性。心电图:窦性心律,T波异常；颈动脉超声:左侧椎动脉血流峰值流速减低。甲状腺B超:甲状腺弥漫性病变。脑电图检查:轻度异常脑电图,各区可见少量6～7 Hz θ波,颞部可见少量5～6 Hz θ波；睡眠脑电图:不正常脑地形图,背景活动中,各区可见少量5～7 Hz θ波,进入睡眠二期中,左侧半球可见一次尖波发放,双侧颞部可见一次尖波发放。心脏彩超:心内结构及血流未见明显异常,左室收缩功能正常。肌电图检查:未见明显异常。头部MRI检查:未见明显异常。腹部B超:肝囊肿。简易精神科智能量表(MMSE):28/30,蒙特利尔认知评估量表(MOCA):16/30,提示中度认知功能减退。2011年10月28日甲状腺功能:三碘甲状原氨酸 0.33(0.58～1.59) ng/ml,甲状腺素16.3(48.7～117.2) μg /L,促甲状腺素>100.00 (0.35～4.94) μIU/ml,游离甲状腺素<4.0(7.0～14.8) ng/L,游离三碘甲状腺原氨酸1.07(1.71～3.71) pg/ml；甲状腺相关抗体: 甲状腺球蛋白抗体 >1 000.00(0～4.11) IU/ml,甲状腺过氧化物抗体 > 1 000.00(0～5.61) IU/ml; 反三碘甲状腺原氨酸 0.67 (0.6～1.81) ng/ml。2011年10月28日脑脊液常规检查:无色透明,比重1.010,细胞计数4.00×106/L,红细胞0/HP, Pandy试验微量。葡萄糖 4.61(2.2～3.9) mmol/L,脑脊液蛋白 545(150～450) mg/L,氯 126.3(120～132) mmol/L。血脑屏障破坏,但未见伴鞘内IgG合成增加。入院后给予吡拉西坦静滴改善脑功能,请内分泌科会诊,确诊为原发性甲状腺功能减退,予左甲状腺素钠片50 μg 每日一次口服,患者临床症状缓解,口齿不清明显改善。考虑为桥本脑病。患者拒绝服用皮质类固醇激素。2011年11月7日复查甲状腺功能,各项指标较前好转,甲状腺球蛋白抗体滴度水平较前有所下降,好转后出院。随访截至发稿前口齿不清未再发作。

Abstract:1 病例资料患者,女,45岁,2008年7月开始出现口干、多饮、多尿现象,半年内体质量减轻9 kg。2009年1月无诱因出现畏寒、发热,最高体温达39℃,伴纳差、乏力、恶心及双膝关节疼痛；空腹血糖为31.2 mmol/L,尿酮体阳性,白细胞(WBC) 12.6×109/L。于外院接受胰岛素静滴及头孢替安抗感染治疗7 d,但膝关节肿痛进行性加重,双下肢逐渐出现凹陷性水肿,右足背第3、4趾处发生皮肤剥脱,局部红肿破溃(图1)。于2009年1月21日转入我院,入院时测空腹血糖17 mmol/L,外周血WBC 23.1×109/L,中性粒细胞0.88,血红蛋白(Hb) 85 g/L,白蛋白21 g/L,血钠133 mmol/L,血钾3.4 mmol/L,诊断为2型糖尿病。予口服二甲双胍治疗,症状无明显好转。