Purpose To study the expression of HK2 in human prostate cancer (PCa) tissues and its effect on malignant phenotype of prostate cancer cells.Methods HK2 expression in PCa tissues was determined by microarray database and immunohistochemical staining.Subsequently,the change of cellular phenotype was detected by glycometabolism kit,CCK-8 kit,and flow cytometry after HK2 knockdown.Results HK2 expression was elevated followed by prostate cancer development.HK2 depletion inhibited cellular proliferation and aerobic glycolysis,and increased the ratio of early apoptosis.Conclusion HK2 expression increases in the process of PCa malignant progression.It plays a critical role in cellular proliferation,glycometabolism,and apoptosis,the mechanism of which needs further exploration.

Objective To study the effect of age on the recurrence-free survival rate after hepatic resection for hepatocellular carcinoma(HCC)and the relationship between microvessel density (MVD)and recurrence of HCC in the elderly. Methods　Severty one cases of elderly patients with HCC were analyzed retrospectively with 352 cases of non-elderly HCC patients as control,and the effect of age on the recurrence-free survival rate was studied.The expressions of CD34 and endocan in HCC tissues were detected by immunohistochemistry in 30 elderly and 30 non-elderly patients.Results　The 1-,3- and 5-year recurrence free survival rates were 75.7%,43.0% and 43.0% in the elderly group respectively,which were higher than those in the non-elderly group(53.6%,38.5% and 33.4%,respectively,Log Rank value=10.25,P＜0.05).The positive rate of alpha fetoprotein (AFP)in the elderly group was 47.9%,which was lower than that in the non-elderly group(62.2%)(X2=23.68,P＜0.05).The median survival times in the high CD34-MVD group and high endocan MVD group were shorter than those in the low CD34-MVD group and low endocan-MVD group(260 d vs.850 d,360 d vs.800 d,Log Rank value was 22.18 and 20.56 respectively,both P＜0.05).Conclusions　The long-term prognosis of hepatic resection for HCC is better in elderly patients than in non-elderly patients.The recurrence of HCC in the elderly is closely related with angiogenesis.

Objective To explore the diagnostic value of carcinoembryonic antigen(CEA),adiponectin combined with leptin in colorectal cancer.Methods The case control study was adopted.Forty cases newly diagnosed as colorectal cancer were selected.Forty cases of non-colorectal cancer were selected according to 1 ∶ 1 matching proportion.Serum levels of CEA,adiponectin and leptin were measured in the study subjects.The ROC curve was used to evaluate the diagnostic value of adiponectin combined with leptin for colorectal cancer.Results Serum level of adiponectin in the experimental group was lower than that in the control group,while the CEA and leptin levels were higher than those in the control group,the difference between the two groups was statistically significant(P＜0.05).The area under the curve of adiponectin combined with leptin reached 0.714±-0.058,its specificity and sensitivity were 77.5 ％ and 65.7 ％ respectively.The area under the curve of adiponectin,leptin combined with CEA reached 0.818 ±-0.048,its specificity and sensitivity were 82.5 ％ and 89.5 ％ respectively.The cut-off values of leptin and adiponectin were 11.53 μg/mL and 9.08μg/mL respectively.Conclusion The combined screening of CEA,adiponectin and leptin can significantly improve the detection rate of colorectal cancer.

Objective To determine the clinicopathologic characteristics of mucinous borderline ovarian tumors (MBOT) and evaluate the risk factors for recurrence. Methods A retrospective study included age, the level of Preoperative serum CA125, surgical procedures, surgical-staging and the risk factors for recurrence in 130 patients with MBOT who were treated from Jan. 1994 to Dec. 2008 in Peking Union Medical College Hospital was done. Results Preoperative serum CA125 and CA199 were elevated in 34% (33/96) and 50% (13/26) of patients respectively. Fifty-two radical surgeries included total hysterectomy and bilateral saipingo-oovarectomy (THBSO) and 78 fertility-sparing surgeries included 54salpingo-oovarectomies (SO) and 24 cystectomy were done. Fifty-five cases underwent comprehensive surgical staging. Mean size of the tumors was (16 ± 10)cm and 90. 0% (117/130) were limited to unilateral ovary. There were 59 (45.4%) cases, 62 (47.7%o) cases, 2 (1.5%) cases and 7 (5.4%) cases in stage Ⅰa, Ⅰc, Ⅱ , Ⅲ , respectively. Forty-five(34.6%)concurrent with benign mucinous tumors, 14(10.8%)ovarian intraepithelial carcinoma, 8 (6.2%) micro-invasive carcinoma and 4 (3.1%) pseudomyxoma peritonei were found. Median duration for follow-up was 56.3 months. Sixteen (12.3%) recurrences and 2 tumor related deaths were found. Median duration from surgery to recurrence was 25.6 months. Recurrent rate after THBSO(4%, 2/49)was significantly lower than that of SO(13%, 7/54) and cystectomy (17%,4/24; P ＜ 0.05). The recurrent rate of Ⅰc or Ⅲ was 18% (11/62) or 3/7, which were significantly higher than that of stage Ⅰa (3% ,2/59; P ＜0.05). Three of the 4 pseudomyxoma peritonei appeared recurrence.While,the results showed that these were no effect on recurrent rate whether concurrent intraepithelial,microinvasive carcinoma or not comprehensive staging surgery. Conclusions Majority of MBOT were diagnosed in early stage and have favorable prognosis. Patients who take conservative surgery had higher recurrence rate than those radical surgery, but it doesn't affect survival. Late stage and concurrent pseudomyxoma peritonei are risk factors for recurrence.

BACKGROUND: Previous studies have confirmed that in the animal models of articular cartilage defects and osteoarthritis, the chondrocytes can overexpress the matrix metal oproteinases. Various abnormal stimuli are likely to break the balance between matrix metal oproteinase and tissue inhibitor of metal oproteinase, thus leading to degeneration of extracel ular matrix of articular cartilage, as wel as the decline and offset of cartilage chondrocytes. OBJECTIVE: To observe the effect of cyclic tensile strain on the expression of matrix metal oproteinases during the repairing process of rabbit articular cartilage defects. METHODS: The animal models of articular cartilage defects were established, and chondrocytes were separated for culture at 10 weeks after operation. The chondrocytes on the non-surgical side were considered as the normal group, and the chondrocytes on the surgical side were randomly divided into high cyclic tensile strain group, low cyclic tensile strains group and control group, and the load amplitude was sin10%. Then 0.1, 1.0 and 0 Hz cyclic tensile strains were loaded respectively. The expressions of matrix metal oproteinases 2, 3, 9 and 13 in each group were detected with reverse transcription-PCR at 24, 48 hours, 1, 2 and 4 weeks after loading cyclic tensile strain. RESULTS AND CONCLUSION: There were significant differences in the expressions of matrix metal oproteinases 2, 3, 9 and 13 at 24 hours after loading cyclic tensile strain between the normal group and the control group (P < 0.05); and there were significant differences in the expressions between the high cyclic tensile strain group and the low cyclic tensile strain group at 1, 2 and 4 weeks after loading cyclic tensile strain (P < 0.05).At the same time, the expressions of matrix metal oproteinases 2, 3, 9 and 13 in the low cyclic tensile strain group were continued to decline, and there were significant differences in the expressions after loading cyclic tensile strain for 24 hours and 4 weeks (P < 0.05). The results indicate that mechanical load can affect the expression of matrix metal oproteinases in the healing process of rabbit articular cartilage defects. In the cel ular and molecular level, the incidence and development of pathological articular cartilage defect and stress should affect each other.

Objective To evaluate the effect of electroacupuncture (EA) pretreatment on NODlike receptor pyrin domain-containing 3 (NLRP3) in neurons during cerebral ischemia-reperfusion (I/R) in rats.Methods Fifty-four adult male Sprage-Dawley rats,aged 7 weeks,weighing 250-280 g,were randomly divided into 3 groups (n =18 each) using a random number table:sham operation group (group S),group I/R,and EA pretreatment group (group E).Cerebral I/R was induced by occlusion of the right middle cerebral artery for 90 min using a nylon thread inserted into the internal carotid artery and advanced intracranially to block the blood flow,followed by reperfusion.In group E,the acupoint Baihui was stimulated with an electric stimulator (sparse-dense wave,frequency 2 Hz/15 Hz,intensity ≤ 1 mA) for 30 min once a day for 5 consecutive days,and the model of cerebral I/R was established at 24 h after the last stimulation.At 72 h of reperfusion,neurological function was assessed and scored.The rats were then sacrificed,and their brains were removed for determination of cerebral infarct volume (using TTC staining),expression of NLRP3,caspase-1 and interleukin-1beta (IL-1β) in brain tissues (by Western blot),and expression of NLRP3 protein in neurons (by immunofluorescence histochemistry).The percentage of cerebral infarct volume was calculated.Results Compared with group S,the percentage of cerebral infarct volume and neurological scores were significantly decreased,and the expression of NLRP3,caspase-1 and IL-1β in brain tissues was significantly up-regulated in group I/R (P＜0.05).Compared with group I/R,the percentage of cerebral infarct volume and neurological scores were significantly increased,and the expression of NLRP3,caspase-1 and IL-1β in brain tissues was significantly down-regulated ingroup E (P＜0.05).Conclusion The mechanism by which EA pretreatment reduces inflammatory responses during cerebral I/R injury may be related to down-regulation of NLRP3 expression in neurons in rats.

The regulation mechanism of arecoline on rat hepatic CYP2E1 was studied in vivo. After oral administration of arecoline hydrobromide (AH; 4, 20 and 100 mg x kg(-1) x d(-1)) to rats for one week, the hepatic CYP2E1 mRNA level remained unchanged, but the hepatic CYP2E1 protein content was dose-dependently increased. Additionally, although the hepatic CYP2E1 activity was induced by AH treatment, the induction was attenuated with the increase in dosage. The results indicate that the effect of arecoline on rat hepaticdoes not involve transcriptional activation of the gene, but largely involves the stabilization of CYP2E1 protein against degradation or increased efficiency of CYP2E1 mRNA translation, and additionally involve the post- ranslational modification of CYP2E1 protein. Furthermore, the CYP2E1 response is fairly equal among the different species, the induction of rat hepatic CYP2E1 by arecoline suggests that there is a risk of metabolic interaction among the substrate drugs of CYP2E1 in betel-quid use human.
Animals ; Arecoline ; pharmacology ; Cytochrome P-450 CYP2E1 ; metabolism ; Cytochrome P-450 CYP2E1 Inducers ; pharmacology ; Humans ; Liver ; drug effects ; metabolism ; RNA, Messenger ; Rats

Objective To construct a recombinant expressing plasmid of the hy1 gene of Enterococcus faecium and to express the recombinant Hy1 protein in E.coil.To explore the immune response in mice fed orally with Hyl protein.Methods hy1 gene was amplified by polymerase chain reaction(PCR)and inserted into a prokaryotic expression vector pQE-30.The recomhinant plasmids were transfected into DH5_?to express Hy1 fusion proteins,which were purified by Ni~--column. Western blot was employed to confirm the immunogenicity of the purified protein.Mice were immu- nized by feeding with the fusion protein.The concentrations of antigen-specific antibody in the serum, mucosal fluid and faces were detected by enzyme-linked immunosorbent assay(ELISA).The role of these antibodies in the anti-infection response was evaluated after the mice were challenged with TX0016.Results hy1 gene was sequenced as 1662 bp,the fusion protein encoding polypeptides of 553 amino acid residues.The relative molecular weight was 60 000 when it was determined by sodium dodecylsulfatepo-lyacry-lamide gel electropboresis(SDS-PAGE).The dissolvable expression protein accounted for 38% of total cell protein.After processed by affinity chromatography,the purity of fusion protein was above 92%.Western blot analysis confirmed that fusion protein could be specifically recognized by the anti-TX0016 serum.The concentrations of serum IgA,serum IgG,faeces sIgA and intestmucosal fluid sIgA was 0.365?0.048,0.431?0.064,0.743?0.056 and 1.112?0.113 respectively in hy1 groups and 0.051?0.013,0.098?0.019,0.102?0.032 and 0.187?0.051 respectively in control group.The differences were statistically significant.The mice survival rate after TX0016 challenge was 70% in hyl group and 50% in control group.There was significant difference between these two groups.Conclusion The results indicate that oral immunization with hyl can induce effective mueosal immune response and produce high level sIgA.

Recent studies have demonstrated that five subtypes (M1-M5) of muscarinic acetylcholine receptor (mAChR) are expressed in the vestibular periphery. However, the exact cellular location of the mAChRs is not clear. In this study, we investigated whether there is the expression of M1-M5 muscarinic receptor mRNA in isolated type II vestibular hair cells of guinea pig by using single-cell RT-PCR. In vestibular end-organ, cDNA of the expected size was obtained by RT-PCR. Moreover, mRNA was identified by RT-PCR from individually isolated type II vestibular hair cells (single-cell RT-PCR). Sequence analysis confirmed that the products were M1-M5 mAChR. These results demonstrated that M1-M5 mAChR was expressed in the type II vestibular hair cells of the guinea pig, which lends further support for the role of M1-M5 mAChR as a mediator of efferent cholinergic signalling pathway in vestibular hair cells.

Objective To investigate the adiponectin(ADIPOQ)gene polymorphisms and which association with colorectal cancer(CRC).Methods Genotyping of blood samples were performed for 250 case-control pairs.Taqman real-time PCR was used to test the three single nucleotide polymorphisms(SNPs),namely rs266729,rs2441 766,rs1 501299.Logistic regression was applied to assess the effects of three SNPs,the gene-gene,and gene-environment interactions on CRC risk.Results The ADIPOQ rs266729 GG and CG+GG genotype had a higher CRC risk than those carrying the CC genotype[OR (95%CI ):1.87 (1.01 - 3.47),1.63 (1.14-2.32),respectively].The same results was observed in cases who carried TG+GG genotype vs .TT genotype in rs2441 766 [OR(95%CI ):1.45(1.02-2.06)].The rs1 501299 GT+TT genotype had a lower CRC risk than those carrying the GG genotype [OR(95%CI ):0.61(0.43-0.88)].Furthermore,in two-factor gene-environment interaction analyses,rs266729 presented signifi-cant interactions with Body mass index (BMI),with OR of 1.1 6 (95%CI :1.03-1.30).Conclusion The results suggest that vari-ants in ADIPOQ may contribute to increased colorectal cancer risk and this contribution may be modified by BMI.