The Nedd8-activating enzyme inhibitor MLN4924 suppresses colon cancer cell growth via triggering autophagy

Neddylation is a post-translational protein modification process. MLN4924 is a newly discovered pharmaceutical neddylation inhibitor that suppresses cancer growth with several cancer types. In our study, we first investigated the effect of MLN4924 on colon cancer cells (HCT116 and HT29). MLN4924 significantly inhibited the neddylation of cullin-1 and colon cancer cell growth in a time and dose-dependent manner. MLN4924 induced G2/M cell cycle arrest and apoptosis in HCT116 and HT29 cells. Moreover, MLN4924 also triggered autophagy in HCT116 and HT29 cells via suppressing the PI3K/AKT/mTOR pathway. Inhibiting autophagy by autophagy inhibitor 3-MA or ATG5 knockdown reversed the function of MLN4924 in suppressing colon cancer cell growth and cell death. Interestingly, MLN4924 suppresses colon cell growth in a xenograft model. Together, our finding revealed that blocking neddylation is an attractive colon cancer therapy strategy, and autophagy might act as a novel anti-cancer mechanism for the treatment of colon cancer by MLN4924.

Establishment of an intestinal organoid-based method to investigate the radiation mitigation effect of Epiregulin / 中华放射医学与防护杂志

Objective To establish an intestinal organoid-based assay to investigate the radiation mitigation effect of epiregulin in vitro. Methods Intestinal crypts were released from tissue incubated with EDTA. Intestinal crypts seeded in 3D matrigel were irradiated at 24 h after plating. The radiation mitigation effect of epiregulin was evaluated by measuring the survival rate, size and budding numbers of the organoid after irradiation, and the basic FGF was used as a positive control of epiregulin. Results Radiation-induced lethality and dose-dependent survival curve of the intestinal organoid were consistent with in vivo data. Treatment with epiregulin (400 ng/ml) at 24 h post-radiation significantly increased survival rate of 8 Gy X-ray irradiated intestinal organoid in comparison with non-treated group [(12.56 ± 1.02)％vs. (4.73 ± 0.38)％, t=12.43,P<0.05]. Conclusions Epiregulin has radiation mitigation effect on intestinal organoid and could serve as a potential medical countermeasure to mitigate gastrointestinal toxicity.

Comparative study of clinical efficacy between the Solitaire mechanical thrombectomy and selective arterial thrombolysis in treatment of acute cerebral infarction / 局解手术学杂志

Objective To investigate the clinical safety and efficacy of mechanical thrombectomy(MT) with Solitaire stent compared with the selective intra-arterial thrombolysis (IAT) in the treatment of acute cerebral infarction.Methods Totally 82 cases patients with severe acute ischemic stroke caused by middle cerebral artery stenosis from January 2014 to May 2016 in the stroke treatment center of the first people's Hospital in ZunYi city who were applied with the mechanical thrombectomy or the selective intra-arterial thrombolysis was included,and a comparative analysis was conducted on the mTICI rating to assess the interventional recanalization,the NIHSS score after the treatment,bleeding rate,and conditions of neurological functional recovery 90 days after operation.Results There were 42 cases applied with the mechanical thrombectomy (MT group),40 cases applied with the intra-arterial thrombolysis (IAT group).The total effective rate reached 85.70％ in the MT group and 62.50％ in the IAT group,the difference was significant (P ＜ 0.05).Compared with NIHSS score before operation,the score after the treatment showed a decreased trend.And the NIHSS score of MT group was better than that of the IAT group in a week after the treatment (P ＜ 0.05).The bleeding rate was lower in the MT group with statistically significant difference (P ＜ 0.05).Conclusion Compared to IAT,MT can provide broader time window,higher recanalization rate and better outcome in patients with severe acute ischemic stroke.

Surgical resection combined with single-stage patch repair for huge desmoid tumor of abdominal wall with intestinal tube / 中华消化外科杂志

Desmoid tumor of abdominal wall was also known as fibronmatosis of abdominal wall.It is a rare benign fibroblastic tumor, which is characterized by non-enveloped,strong local infiltration, with high recurrence rate after surgery.In December 2011, one patient with huge desmoid tumor of abdominal wall with intestinal tube received surgical resection with single-stage patch repair for the large defect in abdominal wall at the 210th Hospital of PLA, and the result was satisfactory.

Diagnosis and treatment of primary appendiceal mucinous adenocarcinoma / 中华消化外科杂志

Appendiceal mucinous adenocarcinoma is a rare disease.The preoperative diagnosis is almostly impossible due to the lack of typical symptoms and inexperienced surgeons.One patient with appendiceal mucinous adenocarcinoma was diagnosed successfully at the 210th Hospital of Chinese PLA,who was misdiagnosed as with periappendiceal abscess by other hospitals.The result of intraoperative frozen pathological section confirmed appendiceal mucinous adenocarcinoma.And then the patient received extended resection and effective recovery.

The effects of fluorine on expression of osteoprotegerin and receptor activator of NF-κB ligand in cultured rat osteoblasts / 中华地方病学杂志

Objective To study the influence of fluorine on signaling pathway of osteoprotegerin(OPG)/ receptor activator of NF-κB ligand(RANKL) in cultured rat osteoblasts.Methods Osteoblasts were isolated from skull of neonatal rats(＜ 24 hours) by enzyme digestion,and fluorine of different concentrations [0 (control),1 × 10-3,1 × 10-4,1 × 10-5,1 × l0-6 and 1 × 10-7 mol/L] were added into the culture medium of second generation of osteoblasts.The expressions of OPG and RANKL mRNA were determined using real-time PCR 24 and 48 hours after culturing.The expressions of OPG and RANKL protein were measured by Western blotting.Results ① After exposed to fluorine for 24 hours,the differences of RANKL and OPG mRNA expression had statistical significance between groups(F =30.95,22.62,all P ＜ 0.01),the expression of RANKL mRNA(5.99 ± 0.39) in the 1 × 10-5 mol/L group and the expressions of OPG mRNA(3.52 ± 0.09,4.81 ± 0.15,3.68 ± 0.04) in the 1 × 10-4,1 × 10-5 and 1 × 10-6 mol/L groups were higher than those of the control group(3.20 ± 0.19,3.09 ± 0.58,all P ＜ 0.05),but in the 1 × 10-3 mol/L group,RANKL mRNA(2.29 ± 0.18) was lower than that of the control group(P ＜ 0.05).After exposed to fluorine for 48 hours,the differences of RANKL and OPG mRNA expression had statistical significance between groups(F =26.62,5.72,all P ＜ 0.01),the expressions of RANKL and OPG mRNA(6.67 ± 0.49 and 5.05 ± 0.51) in the 1 × 10-5 mol/L group were higher than those of the control group(4.29 ± 0.07 and 4.34 ± 0.12,all P ＜ 0.05),and in the 1 × 10-3 mol/L group the expression of OPG mRNA(3.63 ± 0.49) was lower than that of the control group(P ＜ 0.05).② The expression of RANKL protein was not statistically significant between 24 hours and 48 hours groups (F =0.07,0.49,all P ＞ 0.05) ; the differences of OPG protein expression had statistical significance between groups(F =3.26,P ＜ 0.05),the expression of OPG protein in the 1 × 10-5 mol/L group(1.45 ± 0.10) was higher than that of the control group(1.05 ± 0.06,P ＜ 0.05) at the 24 hours.After 48 hours,the expression of OPG protein was not statistically significant(F =0.44,P ＞ 0.05).Conclusions At lower fluorine concentrations,bone formation is the main activity.But when fluorine concentration increased and time prolonged,the osteoclast differentiation and maturation are promoted,and the bone resorption is the main thing.

Strategies of surgical approaches of local resection for mid-lower rectal tumors and pelvic floor neoplasia / 国际外科学杂志

Objective To investigate the strategies of surgical approaches,indications and surgical techniques of local resection for mid-lower rectal tumors and pelvic floor neoplasia.Methods Clinical data of 122 patients underwent local resection for mid-lower rectal tumors pelvic floor neoplasia between July 2004 and July 2008 were analyzed retrospectively.Results Transanal,transsacral,transsphincteric local resection was respectively performed in 45,and 32,and 45 patients.Pathological examination proved that benign tumors were account for 81 cases,pelvic floor neoplasia 16 cases,malignant tumors 25 cases.The masses were 5.6 cm(0 to 12 cm) apart from the anal border,and the mean tumors' diameter was 4.2 cm (0.5 to 11 cm).No case was diagnosed with positive margins upon final pathology of resected specimens.The rate of postoperative complications of transanal,transsacral,transsphincteric approaches was 8.9％ (4/45),18.8％ (6/32),20.0％ (9/45),respectively.The recurrence of transanal,transsacral,transsphincteric approaches was 6.7％ (3/45),9.4％ (3/32),4.4％ (2/45),respectively.Conclusions The three approaches for patients suffering from mid-lower rectal tumors and pelvic floor neoplasia have respectively advantages.Transsphincteric approach is the most useful methods,but with more postoperative complications,so it need more surgical techniques.

Estrogen modulates the inhibited estrogen receptor (ER) expression and the stimulated peroxisome proliferator-activated receptor γ (PPARγ) expression by daidzein (DA) in cultured osteoblasts / 复旦学报（医学版）

Objective To explore the effects of daidzein (DA) on the expressions of estrogen receptors (ER) and peroxisome proliferator-activated recepor γ (PPARγ) in osteoblasts and the influence of estrogen on these effects.Methods A mouse osteoblastic cell line MC3T3-E1 cultured in α-MEM containing 2％ FBS was treated by 0.1 and 10 μmol/L DA.ER antagonist ICI182780 and PPARγ antagonist GW9662 in 0.1 μmol/L was added as required,and an equivalent amount of phosphate buffer solution (PBS) was used as control.For the study on estrogen effect,the cells were treated by DA in the serum-free medium with or without 10 nmol/L 17β-estradiol (E2).The expressions of ERa,ERβ and PPARγ were determined by real-time RT-PCR and Western blot analysis,respectively.Results DA inhibited ER,expression but stimulated PPARγ expression in the cells at the concentration of 0.1 and 10 μmol/L.The down-regulation of ERα by DA could be blocked by ICI182780,whereas the up-regulation of PPARγ could be repressed by GW9662 in transcription levels.Furthermore,the inhibitory effect of DA on ERβ expression was markedly enhanced,while its stimulatory effect on PPARγ expression was almost lost in serum-free medium with 10 nmol/L 17βestradiol as determined by real-time RT-PCR.Conclusions Besides its direct roles in ERs and PPARγ mediated gene transcriptions,DA could exert indirect effect on cellular pharmacological responses by altering ER and PPARγ expressions.The predominant influence on receptors expression probably involved in the time-related biphasic effects of DA on osteogenesis,which was supposedly influenced by estrogen level.

Effects of daidzein on steroid receptor coactivator-1 expression in MC3T3-E1 cells and the mechanism / 中西医结合学报

To investigate the roles of daidzein in the expressions of steroid receptor coactivator-1 (SRC-1) and nuclear receptor corepressor (NcoR) in MC3T3-E1 osteoblastic cells.

Detecting optimal cut-off value of international normalized ration by receiver operator characteristic curve in diagnosis of hemorrhage in patients with nonvalvular atrial fibrillation receiving wafarin / 中华老年医学杂志

Objective To evaluate the value of international normalized ratio (INR) in predicting and diagnosing hemorrhagic events in patients with nonvalvular atrial fibrillation (NVAF)receiving wafarin, to determine the optimal cut-off value of INR for predicting hemorrhagic events by receiver operator characteristic (ROC) curve. Methods The data of 231 patients with NVAF receiving wafarin were retrospectively analyzed, including 93 patients with hemorrhagic events and 138cases without hemorrhagic events as control group. The PT and INR were detected by Sysmex CA-500 with Medcalc software plotting ROC curve. Results The area under the ROC curve for INR was 0. 822 (95%CI: 0. 717-0. 900), the analysis of ROC curve revealed the optimal cut-off value of INR was 2.71, which presented a sensitivity of 77.40% and a specificity of 78.30%. Conclusions Through evaluation by ROC curve, the new cut-off value provides substantial improvement in sensitivity, with an acceptable loss of specificity. The value on predicting hemorrhagic events is better, the discriminative power of INR between hemorrhagic events and nonhemorrhagic events is satisfactory. The cut-off value of INR can guide the clinical physicians to predict the risks of hemorrhagic and promote proper use of clinical medications.

Fibroblast growth factor 23 expression in rat calvarial osteoblastic cells and its responses to hormones in vitro / 复旦学报（医学版）

Objective To investigate the expression patterns of fibroblast growth factor 23 (FGF23) in osteoblast and its responses to calcium, phosphate, exogenous PTH and 1,25(OH)_2D~3. Methods The primary rat calvarial osteoblasts were cultured in MEM medium which containing 10% FBS, then were harvested when cells were in half-confluence, confluence, osteoid deposition and osteoid mineralization stages respectively. The procedure was monitored under microscopy. Total RNA was extracted from cells according to the Trizol procedure. FGF23 mRNA levels were determined by Real-time PCR. Further, the confluent osteoblasts were treated with 3.2 mmol/L CaCl_2, 4.4 mmol/L β-glycerophosphate, 10~(-9) mol/L rhPTH(1-34) and 10~(-8) mol/L 1,25(OH)_2D_3 respectively for 3 days, and same volume of the medium was added as the control. The gene expressions were determined by Real-time PCR. Results FGF23 expression was transiently up-regulated at cell confluent stage and down-regulated after that. The FGF23 mRNA levels were 7.5-fold higher in confluent cells compared with that in half-confluent cells (P<0.001). The markedlly stimulating effect (about 16 times) on FGF23 expression was stimulated by exogenous 1,25(OH)_2D_3 treatment while no significant effect was found on FGF23 mRNA levels by CaCl_2,β-glycerophosphate, and rhPTH(1-34) treatments when compared with the control. Conclusions The FGF23 expression in osteoblast is developmental stage-related and its powerful stimulator is 1,25(OH)_2D.

Earlier pathological character and mechanisms of irradiation necrosis on femoral head / 中华放射医学与防护杂志

Objective To observe the earlier pathological character and mechanism of radiation osteonecrosis in femoral head, in order to provide evidences for the earlier diagnosis and prevention of radiation osteonecrosis of femoral head. Methods Single femoral head of rats were irradiated singly with 30 Gy of 137 Cs γ-ray. The rats were executed after 2, 6 and 12 weeks, then the femurs were stained with HE and histopatholngical changes were observed by light microscope. The bone marrow mesenchymal stem cells (BMSCs) were cultured after 2 weeks and its proliferation and the colony formation were observed. The rats were endo-perfused with microfili contrast medium 12 weeks later, and the 3-dimensional structure of capillaries by Micro-CT was re-estabhshed to detect the pathological changes of capillaries after irradiation. Result The irradiated femur showed deranged cbondrocyte, decreased osteocyte, shrinking nucleus, increased empty bone lacuna and reduced bone trabocnla (P < 0.05). Micro-CT showed the discontinued small vessels and absence(6.65 %) capillaries in irradiated femur were obviously less than those of the unirradiated (12.3 %)(P < 0.001). The proliferation of BMSCs was slowed, the number of colony in irradiated group (10 %) was less than that of control (21 %) (P < 0.001). Conclusions The preliminary histopathological changes of osteoradionecrosis on femoral head could be increased the empty bone lacuna, and the bone lacuna above 30 % was the sign of the earlier period of osteoradionecrosis. The osteonecrosis of femoral head induced by radiation is not only correlated to the damages to the bone, but also to the damages to BMSCs and capillaries.

Interventional effect of ibandronate on osteoporosis in ovariectomized rats / 中国组织工程研究

BACKGROUND: Diphosphonate has a predominant therapeutic effect in the prevention and treatment of postmenopausal osteoporosis. Ibandronate, as a new-type diphosphonate preparation, is gradually becoming a study hotspot.OBJECTIVE: This study is to investigate the efficiency of ibandronate in interfering postmenopausal osteoposis by observing bone mass loss related indexes in ovariectomized rats, and made a comparison with nilestriol.DESIGN: A completely randomized grouping, and controlled animal experiment.SETTING: Study Room for Bone metabolism, Fudan University Medical College.MATERIALS: Forty SD female rats, aged 10-12 months, were involved in this study. Ibandronate was provided by the State Key Laboratory of Jiangsu Institute of Atomic Medicine. Nilestriol was produced in the Shanghai 12th Pharmaceutical Factory.METHODS: This experiment was carried out in the Study Room for Bone Metabolism, Institute of Radiation Medicine,Shanghai Medical University between August 1996 and June 1998. The rats were divided into 4 groups by a lot, 10 rats in each: sham-operation group, ovariectomized group, ovariectomized+ibandronate group and ovariectomized +nilestriol group. In the sham-operation group, only small pieces of adipose tissue around the ovary were resected from the rats.Three months after operation, each rat was intragastrically administrated with 1 mL normal saline; In the ovariectomized group, ovariectomized+ibandronate group and ovariectomized +nilestriol group, each rat was subjected to bilateral ovariectomy, and 3 months later, they were intragastrically administrated with normal saline, ibandronate water solution [0.5 mg/( kg·d)] and nilestriol suspension [1 mg/(kg· time)] respectively. Each rat in the latter three groups was administrated for 90 days, twice in the first week, and then once a week.MAIN OUTCOME MEASURES: Left femur was taken out, and its dry weight and ash weight were measured. Calcium content of bone was determined with an atomic absorption spectrophotometer, bone density of the whole body with a bone density apparatus, the bone density at the juncture of 1/2 right femoral bone length with a single photon bone density apparatus, and femoral anti-bending force was determined with a universal testing machine. Alkaline phosphatase were determined by dynamical method with an automatic biochemistry analyzer, urine calcium by EDTA titration method, urine creatinine by picric kinetic method, and urinary hydroxyproline by modified proline assay.Trabecular area was calculated.RESULTS: Forty rats were involved in the final analysis. ① Bone dry weight, bone ash weight and bone calcium content in the ovariectomized group were significantly lower than those in the other 3 groups, respectively (t =13.58-52.98, P ＜0.05). ② Femoral bone density and bone density of the whole body of rats in the ovariectomized group were significantly lower than those in the other 3 groups (t =3.31-5.61, P＜0.05), while anti-bending force was close between ovariectomized group and the other 3 groups (P＞0.05). ③ The ratio of urine calcium to urine creatinine was significantly lower in the ovariectomized+ibandronate group and ovariectomized +nilestriol group than in the ovariectomized group (t =4.04, 3.30, P＜0.05). No significant difference in the alkaline phosphatase and ratio of urinary hydroxyproline to urinary creatinine existed among the groups (P ＞ 0.05). ④Trabecular area of vertebrae in the ovariectomized group was significantly smaller than that in the other 3 groups (t =2.22,2.41,3.45,P ＜ 0.05), while the trabecular area of tibia in the ovariectomized group was only smaller than that in the ovariectomized +nilestriol group (t =2.45, P＜ 0.05).CONCLUSION: Osteoporosis apPears obviously in the SD rats 3 months after ovariectomy. Ibandronate has obviously inhibitory effects on the bone mass loss of rats with postmenopausal osteoposis, and it is equivalent to nilestriol in inhibitory effect.

Low Temperature Steam Formaldehyde Sterilization and Its Application / 中华医院感染学杂志

OBJECTIVE To apply and practice the process and effect of low temperature steam formaldehyde(LTSF) sterilization for heat-sensitive goods.METHODS The heat-sensitive medical instruments were sterilized in LTSF sterilizer under the appropriate media conditions,all procedures,including preparation,packing,sterilization,unloading,storage etc were strictly in the right way.RESULTS Sterile result has been guaranteed.CONCLUSIONS LTSF is a kind of effective sterilization method for low temperature sterilization application.By this way,the quality of sterile goods has been reached,the rate of nosocomial infection has been lowered,requirements of hospital clinical departments for sterile goods also have been met.

Clinical Efficacy on Epinephrine and Vasopressin Combined with Naloxone in the Treatment of Cardiopulmonary Resuscitation / 中国药房

OBJECTIVE:To observe clinical efficacy of epinephrine combined with vasopressin and naloxone for cardiopulmonary resuscitation(CPR)and to investigate its mechanism. METHODS:64 patients with sudden cardiac and respiratory arrest were randomly divided into control group(epinephrine group),treatment group Ⅰ(epinephrine+vasopressin group)and treatment group Ⅱ(vasopressin+epinephrine+naloxone group). Spontaneous circulation,breathing recovery time,restoration of spontaneous circulation and respiration rate,24 h survival rate and discharge survival rate were observed and compared among different groups. RESULTS:As compared with control group,spontaneous circulation,breathing recovery time 24 h survival rate and discharge survival rate in treatment group Ⅰ and Ⅱwere increased significantly(P

Evaluative significance of the expression of Fas antigen and P53 protein in colorectal cancer for the biological characters and prognosis / 中国组织工程研究

BACKGROUND: The expression of Fas antigen and P53 protein in colorectal cancer could reflect the biological character and the prognosis of the tumor.OBJECTIVE: To investigate the relationship between the expression of Fas antigen and P53 protein in coloreetal cancer and the tumor-related biological characters.DESIGN: A controlled experiment by introducing pathological specimens as subjects.SETTING: The study was conducted in the Department of Pathology of Xijing Hospital Affiliated with the Fourth Military Medical University of Chinese PLA.PARTICIPANT S: Totally 32 pathological specimens of colorectal cancer were randomly selected from the Department of Surgery of Xijing Hospital between January 1995 and October 1998. Sixteen normal large intestine tissues were selected form the cadaver of pathology department.METHODS: The expression of Fas antigen and P53 protein in 32 colorectal cancer tissue and 16 normal intestine tissue were tested and compared with immunohistochemical method.MAIN OUTCOME MEASURES: Fas antigen and P53 protein expression in coloreetal cancer and normal intestine, and the clinical indicators of patients with colorectal cancer.RESULTS: The expression of Fas antigen was positive in normal large intestine tissue while the expression of P53 protein was negative. The positive rate of Fas antigen expression in colorectal cancer was 41% (13/32), while the positive rate of P53 expression in colorectal caner was 53% (17/32). Fas antigen significantly decreased in colorectal cancer tissues. The expression of Fas antigen was correlated with the histological type of colorectal cancer and its liver metastasis( P ＜ 0.05) . And the expression of P53 protein was correlated with the liver metastasis of colorectal cancer( P ＜ 0. 05).CONCLUSION: The expression of Fas antigen significantly decreases in colorectal cancer. The higher the malignance is, the more significant less the expression is. Fas antigen does not express in liver metastasis patients. P53protein had positive expression in colorectal cancer tissues, especially in patients with liver metastasis. The reduction and deletion of Fas antigen expression and the excessive expression of P53 protein indicate poor prognosis.

Inhibitory Effect and Cell Mechanism of BM210955 on Bone Resorption / 复旦学报（医学版）

PurposeTo investigate the mechanism of BM210955 (ibandronate, a new bisphosphonate drug manufactured in China)in the prevention of bone loss in vitro. MethodsThe osteoclasts isolated from the long bone of 10-day-old Rabbit were cultured on glass and bone slices in different concentration of BM210955. TRAP(tartrate resistant acid phosphonase) staining for osteoclast and TB( toluidine blue)staining for resorption lacunae were used on bone slices and Fluorescence (orange acridine) staining for apoptosis cell on glass slices was used;Multinucleated( three and more nuclei) TRAP positive cell and Apoptosis cell and Pits were counted. ResultsThe BM210955 decreased the multinucleated cell number by 73% in 10-8 mol/ L;The inhibitory rate of pit formed was correlate to concentration as 31.58%, 76.32% and 87.99% to 10-12, 10-10 and 10-8 mol/L; The apoptosis induction was shown in above 10-8 mol/L, and the apoptosis rate was 62% in 10-4 mol/L. Conclusions Induction of osteoclast apoptosis and decrease of the cell number and inhibition of resorptive ability were the major mechanism for bone loss prevention effect of BM210955 in vitro.