Objective To explore the effect and its mechism of osteoprotegerin (OPG) on human umbilical vein endothelial cells(HUVECs) induced by high glucose.Methods (1) The cultured HUVECs were treated with normal glucose(5.5 mmol/L),high glucose(33 mmol/L),high glucose + OPG(0.5,1,and 2 μg/ml)as well as mannitol(5.5 mmol/L glucose+27.5 mmol/L mannitol) for 48 h,respectively.Flow cytometry assay and Hoechst 33258 staining were used to detect the cell apoptosis.The expression levels of Bcl-2 and Bax protein were measured by western blot analysis.(2) The cultured HUVECs were treated with normal glucose,high glucose,high glucose + OPG (2 μg/ml OPG),high glucose + rapamycin(10 ng/ml rapamycin) as well as high glucose + rapamycin + OPG for 24h,respectively.The expression levels of S6K,p-S6K,4EBP1 and p-4EBP1 protein were measured by western blot analysis.(3)The cultured HUVECs were treated with normal glucose,high glucose,high glucose + OPG(2 μg/ml)for 24 h,respectively.The expression levels of tuberin and p-tuberin protein were measured by western blot analysis.Results (1) Compared with normal glucose group,the apoptosis of HUVECs and the expression level of Bax was dramatically increased,and the expression of Bcl-2 was decreased significantly in high glucose group (P＜0.05).The apoptosis of HUVECs and the expression level of Bax in high glucose + OPG group was significantly lower than that in high glucose group,which was still higher than that in normal glucose group (P＜ 0.05).and the expression of Bcl-2 in high glucose + OPG group was significantly higher than that in high glucose group,which was still lower than that in normal glucose group (P＜0.05).There was no statistically difference between hyperosmolar control group and normal glucose group.(2) Compared with normal glucose group,the expression levels of p-S6K and p-4EBP1 were increased markedly in high glucose group(P＜0.05).The expression levels of p-S6K and p-4EBP1 in high glucose + OPG group were significantly lower than that in high glucose group,which were still higher than that in normal glucose group(P＜0.05).No significant difference was found between high glucose + OPG group and high glucose + rapamycin group.(3) Compared with normal glucose group,the expression level of p-tuberin was increased markedly in high glucose group (P ＜ 0.05).The expression level of p-tuberin in high glucose + OPG group was significantly lower than that in high glucose group,which was still higher than that in normal glucose group (P ＜0.05).Conclusions It suggests that the protective effect and mechanism of OPG on HUVECs cultured with high glucose may be association with tuberin/mTORC1 pathway.

ObjectiveTo evaluate ultrasound diagnosis of occult inguinal hernia in children. MethodsThe contralateral site of 132 established unilateral inguinal hernia cases was checked by ultrasound and 33 outpatient suspected occult inguinal hernia cases underwent B-mode ultrasonography. [WT5”HZ]Results Occult inguinal hernia was diagnosed in 15 cases (11.4%) in the 132 children,and diagnosis of occult inguinal hernia was founded by ultrasound in 17 out of the 33 outpatient cases. No opposite inguinal hernia developed in the ultrasound negative 117 patients during follow up of 2 to 12 months,and hernia developed in one out of the 16 outpatient cases initially judged as negative by ultrasound. ConclusionUltrasonography is useful for the diagnosis of occult inguinal hernia in children.

Objective To investigate the prevalence of orthostatic hypotension in hypertensive patients aged 80 or over and associated risk factors at level A tertiary hospitals.Methods A multicenter cross-sectional study was conducted among very old hypertensive patients (≥ 80 years) at 28level A tertiary hospitals by using questionnaire-based surveys.A total of 1298 hypertensive subjects aged between 80 and 101 years were enrolled in this study.Blood pressures was measured in the supine position and the upright position within three minutes of standing,and 316 patients were assigned to the orthostatic hypotension(OH) group and the other 982 patients to the non-orthostatic hypotension(NOH) group.Additional information was acquired through patients' medical records and the questionnaire.Results The prevalence of OH was 24.3％ (316 cases) There were no significan different betueen OH patienl with NOHones in gender and age.OH patients had higher supine systolic and diastolic blood pressure(P=0.003 and 0.000,respectively),lower standing systolic and diastolic blood pressure(P=0.000 and 0.000,respectively),and higher rates of past coronary heart disease,stroke and renal abnormalities (P =0.037,0.001 and 0.014,respectively) than NOH patients.Logistic regression analysis showed that the prevalence of OH in the patients was positively correlated with supine systolic blood pressure(OR=1.196,CI:1.153-1.242,P=0.000) and supine diastolic blood pressure (OR =1.740,CI:1.602-1.885,P =0.000) and was negatively correlated with standing systolic blood pressure(OR =0.824,CI:0.795-0.855,P=0.000) and standing diastolic blood pressure (OR =0.584,CI:0.539-0.634,P =0.000).Conclusions The prevalence of orthostatic hypotension is high in hypertensive patients aged 80 years or over at level A tertiary hospitals.Poor blood pressure control may increase the risk of orthostatic hypotension.

Objective To evaluate the feasibility of the porous titanium/chitosan/hydroxyapatite (Ti/Ch/HA) composite scaffold as a bone repair substitute.Methods Additive manufacturing (3D printing) technology was used to fabricate porous Ti scaffolds as supporting structures.Chitosan/hydroxyapatite (Ch/HA) sponge was prepared within the macro-pores of Ti scaffolds using freeze drying technology.Thus,a kind of composite porous Ti/Ch/HA scaffold with good cell affinity was obtained.Osteoblastic cells were seeded and cultured in pure porous Ti scaffolds and composite Ti/Ch/HA scaffolds for 7 days.The cellular morphology,seeding efficiency and proliferation were examined and compared between the 2 kinds of scaffolds using scanning electron microscopy (SEM) and MTT assay.Results The SEM examination showed that the macro-pores of Ti/Ch/HA scaffolds were full of the composite sponge structure of Ch/HA,with a micropore size of 50 to 200 μm.Like the pure porous Ti scaffolds,composite Ti/Ch/HA ones have a compressive strength of 168.2 to 192.6 MPa,a yielding strength of 137.1 to 154.1 MPa,and a Young's modulus of 3.21 to 4.51 GPa.After culture for 7 days,a large number of flat cells adhered onto the surface of Ti scaffolds while the cells adhering onto the Ti/Ch/HA composite scaffolds were fusiform.The seeding efficiency of osteoblastic cells in the composite Ti/Ch/HA scaffolds (73.218％ ± 3.748％) was significantly higher than that in the pure porous Ti scaffolds (21.352％ ±4.365％) (P ＜0.05);the OD value of the composite Ti/Ch/HA scaffolds (0.783 ±0.043) was significantly higher than that of the pure porous Ti scaffolds (0.382 ± 0.036) (P ＜ 0.05).Conclusions Ti/Ch/HA composite scaffolds can match human bone in mechanical properties.Compared with pure porous Ti scaffolds,the Ti/Ch/HA composite ones are more suitable for adhesion and proliferation of osteoblasts,making them an ideal kind of bone repair substitute.

ObjectiveTo observe the effects of XinShu parenteral solution (XSPS) on the activation of platelet, the activity of fibrinolysis system after intima denudation of rabbits.Methods20 male Japanese white rabbits (2.5±0.5)kg were randomly divided into the control group and XSPS group .The celiac arterial endothelium of all rabbits were denuded with balloon. Before the operation and 3d, 7d, after balloon denudation, vein blood samples were obtained from each group rabbits for measurement of α granule membrane protein of platelets(GMP-140), tissue-type plasminogen activator(t-PA) and plasminogen activator inhibitor-type Ⅰ(PAI-1).ResultsPlasma GMP-140 and PAI-1 activity obviously elevated after balloon injury, and there was a little elevation in plasma t-PA activity in control group. Activity of plasma GMP-140 in XSPS group remained bottom level after balloon injury, and there was a significant increase in plasma t-PA activity and a marked reduce in PAI-1 activity in XSPS group. There was a notable difference between group B and group C (P<0.05). ConclusionXSPS obviously inhibits platelet activation, and improves fibrinolysis activity after balloon injury.

[Objective] The aim of the present study was to investigate the role of membrane estrogen receptor (mER) mediated pathway in the proliferation and apoptosis of endothelial progenitor cells (EPCs). [Methods] Bone marrow (BM)-derived EPCs were cultured. The cells were divided into different groups, plus or not plus estrogen receptor blocker (ICI 182,780), PI3K inhibitors (LY294002), and NOS inhibitor (L-NAME) to show the effect of E_2-BSA on EPCs. The proliferation of EPCs was determined by MTT and nitric oxide (NO) release was measured by chromatometry. Apoptotic cell death was determined using the Hochest 33258 staining. The expression of phosphorylated eNOS (p-eNOS) were detected by Western blot. [Results] E_2-BSA could increase EPCs proliferation, and this effect was inhibited by estrogen receptor blocker ICI 182,780, thus indicated that mER-initiated membrane signaling pathways were involved in the action of estrogen on EPCs. E_2-BSA increased nitric oxide production and inhibited apoptosis induced by serum withdrawal, and this effect also inhibited by PI3K inhibitor (LY294002), NOS inhibitor (L-NAME)and estrogen receptor blocker(ICI 182,780), thus indicated that PI3K/Akt/NO pathway was involved the effect of estrogen on EPCs apoptosis. Moreover, E_2-BSA treatment increased phosphorylation of eNOS (p-eNOS). PI3K inhibitors (LY294002) also blocked these effects. [Conclusions] The results of present study suggested that mER mediated EPCs proliferation and apoptosis were related to the PI3K/Akt/eNOS pathway.

Objective To explore the reasons and countermeasure of the complications after proce-dure for prolapse and hemorrhoids(PPH).Methods From March 2006 to November 2007,a total of 78 patients with Ⅲ-and Ⅳ-degree hemorrhoids underwent PPH.The reasons and countermeasure of the complications were analyzed retrospectively.Results The average operation time time was(16.8±3.4)rain,the average blood loss was(12.0±2.1)ml,and the average hospitalization was(4.0±2.7)days.Patients were followed up for 1-20 months and(13.4±1.7) months for average,follow-up rate time was 92.3% (72/78).Comptications included urinary retention in 24 cases(33.3%),anastomotic bleedings in 5 cases (6.9%),reopelative hemostasis in 1 case(1.4%),postoperative pain of anal in 7 cases(9.7%),severe pain in 1 case(1.4%),slight encopresis in 1 case(1.4%),recurrence in 1 case(1.4%),2 cases(2.8%)were found stricture in rectum at 2,4 months after the procedure and no case appeared rectovaginal fistula.Con-clusion PPH forⅢ-and Ⅳ degree hemorrhoids is effective with fewer complications,which can be pre-vented by standard and skillful operation.

Objective To prepare Exosomes secreted by mouse hepatoma cell (H_(22)) and heat stressed Exosomes (HS-Exo) derived from heat stress-treated mouse hepatoma cell (H_(22)), in order to study the possible anti-tumor immune mechanism. Methods Exosomes and HS-Exo were purified by serial ultracentrifugation and sucrose density gradiant centrifugation, and were observed and identified by electron microscope. The components and production of the protein and the effects of the host immune response against hepatocellular carcinoma of HS-Exo were observed by using Exosomes as the control. Their immunological factors were detected by Western blot. Lymphocyte proliferation and specific cytotoxic activity of mouse splenic cells were determined by MTT. CD4~+ and CD8~+ lymphocytes infiltration in mouse tumor tissues immunized by both were analysed by immunohistochemical staining. Results HS-Exo was similar in morphology to the Exosomes, the important immune-ralated protein expressed in HS-Exo was increased (P<0.05). HS-Exo immunized mouse group showed more effective inhibition of tumor growth, better-induced lymphocyte proliferation,more significantly enhanced the cytotoxic activity of spleen lymphocytes, as well as a more prominent role in tumor therapy than Exosomes immunized mouse control group (P<0.05). Conclusions Heat stress treatment method for the preparation of HS-Exo was feasible. HS-Exo had a stronger role in the immuneactivity and tumor treatment than control Exosomes.

Objective To investigate the CT features of primary hepatic neuroendocrine carcinoma (PHNEC).Methods The CT findings of 8 patients with PHNEC which was confirmed by surgery and histopathology were analyzed retrospectively.Tumor location,size,shape,margin,density,enhancement patterns and degree,portal vein tumor thrombus,hilar and retroperitoneal lymph node enlargement were studied retrospectively.Results Of 8 patients,there were 5 tumors which were located in the right liver,1 in the left liver,and the remaining 2 in both the left and right livers.The CT value of the mass on plain scanning ranged from 20 to 35 HU.The mean value of CT was (29.6 ± 8.4) HU.Tumors showed low density in 5 patients and slightly lower density in 3.In 5 tumors the boundary was clear and in 3 it was not clear.Uneven mass density and different sizes of necrotic cysts were found in 8 tumors.In one of these tumors,the inside showed honey-comb appearance while small areas of haemorrhage were found in the tumor of another 2 patients.For all the eight patients,there were dilatation of peripheral bile ducts,portal vein tumor thrombus,enlarged lymph nodes or peritoneal effusion in hepatic portal area and peritoneal cavity.In the arterial phase,there was mild to moderate enhancement of the mass with CT value of 38 ～ 65 HU.The average value of CT was (54.8 ± 6.9) HU.In the venous phase,there was mild to moderate continuous enhancement of liver parenchyma,with CT value of 40 ～67 HU.The average value of CT was (61.4 ± 11.6) HU.In the delay phase,a substantial part of the mass was continuously enhanced in 2 patients while there was slow fading of enhancement in 6 patients.The CT value were 36 ～ 57 HU,the average value of CT was (46.6 ± 9.2) HU.In 3 patients,the separations inside the tumor showed enhancement.Conclusion CT features combined with clinical findings might give a hint to the diagnosis of primary neuroendocrine carcinoma of liver.

Objective To investigate the influence of bushenhuoxue drug-containing serum on PI3K and Akt signaling pathway in purified retinal ganglion cell (RGCs) in vitro of Sprague-Dawley (SD) rats,and to explore the protective mechanisms of bushenhuoxue recipe on RGCs.Methods At first,bushenhuoxue drug-containing serum was prepared,and the RGCs of SD rats were purified;after the apoptotic model of pressurized and purified RGCs was established successfully in vitro using open pressure control system,RGCs were dealt with 50 g · L-1,100 g · L-1,200 g · L-1 concentration gradient of bushenhuoxue drug-containing serum.Then the subjected cells were divided into normal culture group (N group),control group (C group),50 g · L-1 bushenhuoxue group (50 g · L-1 BSHX group),100 g · L-1 bushenhuoxue group (100 g · L-1 BSHX group),200 g · L-1 bushenhuoxue group (200 g · L-1 BSHX group).Finally,cell apoptotic rate was detected by Annexin V-FITC/PI staining,while real-time quantitative PCR (qRT-PCR) and Western blot were used to detect the mRNA and protein expression of PI3K and Akt in each group respectively.Results The results of qRT-PCR detection showed that PI3K,Akt mRNA expression level in C group (0.04 ±0.01) was decreased compared with N group (1.00 ± 0.04),and the difference was statistically significant (all P＜0.05),while PI3K,Akt mRNA levels in 50 g · L-1,100 g · L-1 and 200 g · L-1 BXHX group (0.18 ±0.01,0.21 ±0.02,0.22 ±0.01,0.36 ±0.01,0.84 ±0.10,1.07 ± 0.17) were increased compared with the C group,and the difference was statistically significant (all P ＜0.05).The Western blot results of each group showed that PI3K,Akt protein expression level in C group was decreased compared with N group,with statistical difference (all P ＜ 0.05),while PI3 K,Akt protein expression levels in 50 g · L-1,100 g · L-1 and 200 g · L-1 BSHX group were increased compared with C group,with staffstical difference (all P ＜ 0.05).Conclusion Bushenhuoxue drug-containing serum may inhibit the RGCs apoptosis induced by pressure,which may be related to the activation of PBK/Akt signal transduction pathway.