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.

Adolescent ; Child ; Humans ; Male ; Neuroectodermal Tumors, Primitive, Peripheral ; pathology ; Sarcoma, Ewing ; pathology

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] 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 investigate the immunoregulatory ability of human lung-derived mesenchymal stem cells (MSCs) on T cells. Methods MSCs were isolated and cultured from human fetal lung. The immune phenotype was tested by flow cytometry and T lymphocyte proliferation assessed by thymidine incorporation. Results HLA-DR, CD86, and CD80 were not expressed in human lung-derived mesenchymal stem cells. The proliferation of peripherial blood-derived T cells was suppressed and this suppression seemed dependent on the concentration of MSCs. Conclusion Human lung-derived MSCs have been proved to possess immunomodulatory ability.

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 protection mechanism of ulinastatin on bacterial endotoxin-induced acute lung injury. Methods Acute lung injury was induced by Escherichia colilipo-polysaccharide(LPS)5 mg·kg-1·d-1,intratracheally. Twenty SD rats were randomly divided into control group(n=10)and ulinastatin group(n=10). Ulinastatid group received ulinastatin 50 kU·kg-1 ,the control groups received the same amount of 0. 9% sodium chloride solution. Then the expression changes of rat AQP-1 and AQP-5,alveolar wall thickness change and the degree of pulmonary edema were detected. Results After the injection of LPS into the rat,the expression of AQP-1 and AQP-5 in control group were continuously decreased,but those in ulinastatin group decreased were not obvious. The lung wet/dry weight ratio in the control group increased significantly,the not obvious changes in the ulinastain group. The thickness of the alveolar in 24,48,72 h of the control group were(3. 84±0. 68),(6. 32±1. 08),(11. 03±2. 47)μm, respectively,and those in the ulinastian groups were(2. 31±0. 44)(,3. 76±0. 82)(,2. 94±0. 67)μm,respectively. Conclusion The AQP-1 and AQP-5 induced the occurrence of pulmonary edema by changing the cell permeability. Ulinastatin can slow down the process so as to reduce the degree of endotoxin-induced lung injury.