Objective To investigate the regulatory effects of PTEN-PI3K/AKT pathway on the apoptosis of gastric cancer MKN28 cells and the possible mechanisms.Methods The specific sequence of PTEN was transfected to MKN28 cells by eukaryotic expression vector (transfection group),and then vacant vector (negative control group) and PBS (blank control group) were transfected to the MKN28 cells,respectively.The effects of PTEN-PI3K/AKT pathway on the apoptosis of MKN28 cells and the expressions of PI3K,AKT,Caspase-3 and Caspase-9 were investigated.The growth curve and apoptosis of the MKN28 cells were detected by MTT assay and TUNEL staining,respectively,and the protein expression was detected by the Western blot.MKN28 cells which did not transfected by the PTEN were processed by inhibitor of PI3K (LY294002) (treated group),and MKN28 cells in the control group were processed by PBS.The expressions of apoptosis protein and apoptosis-related protein were detected after inhibition of PI3 K.All data were analyzed using the t test.Results The model of over-expression of PTEN was obtained and transfected into MKN28 cells.The survival of MKN28 cells in the transfection group was significantly inhibited in a time-dependant manner ( r =0.938,P ＜ 0.05 ).The mean apoptotic rate of the transfection rate was 27.86％ ± 4.78％,which was significantly higher than 0.01％ ± 0.01％ of the negative control group ( t =9.527,P ＜ 0.05 ).The protein expression of PI3K in the transfection group was 0.25 ± 0.03,which was significantly lower than 0.93 ± 0.16 of the blank control group and 0.96 ± 0.15 of the negative control group (t =7.235,8.883,P ＜ 0.05 ).The protein expression of P-AKT in the transfection group was 0.21 ± 0.03,which was significantly lower than 0.93 ± 0.13 of the blank control group and 0.91 ± 0.12 of the negative control group (t =9.347,9.802,P ＜ 0.05 ).The expressions of Caspase-3 and Caspase-9 of the transfection group were 0.86 ± 0.11 and 0.87 ± 0.12,which were significantly higher than 0.16 ± 0.03 and 0.18 ± 0.04 of the negative control group,and 0.15 ± 0.02 and 0.16 ± 0.03 of the blank control group ( t =10.634,10.999,9.448,9.942,P ＜0.05).The apoptotic rate of the MKN28 cells of the treated group was 28.60％ ± 4.50％,which was significantly higher than 0.12％ ± 0.06％ of the control group (t =10.961,P ＜ 0.05 ).The protein expression of PI3K and P-AKT of the treated group were 0.18 ± 0.02 and 0.11 ± 0.01,which were significantly lower than 0.93 ± 0.14 and 0.90 ± 0.12 of the control group (t =9.186,11.363,P ＜ 0.05 ).The protein expression of PTEN of the treated group was 1.15 ± 0.15,which was significantly higher than 0.21 ± 0.08 of the control group (t =9.577,P ＜ 0.05 ).The relative expressions of Caspase-3 and Caspase-9 of the treated group were 0.86 ± 0.12 and 0.88 ± 0.11,which were significantly higher than 0.25 ± 0.02 and 0.21 ± 0.03 of the control group (t =8.685,10.178,P ＜ 0.05).Conclusions Over-expression of PTEN may enhance the apoptosis of gastric cancer cells through inhibition of PI3K/AKT pathway.Inhibition of PI3K can enhance the expression of PTEN.PI3K and PTEN regulate the apoptosis of gastric cancer cells.

Dendritic Cell Sarcoma, Interdigitating ; Humans ; Lymph Nodes ; Male ; Middle Aged

Cardiomyopathy, Dilated ; pathology ; Cardiomyopathy, Hypertrophic ; pathology ; Diagnosis, Differential ; Fetal Diseases ; pathology ; Fetus ; Heart Neoplasms ; pathology ; Humans ; Myocardium ; pathology ; Ventricular Dysfunction, Left ; pathology

To investigate the effect of naringenin on ovariectomy-induced postmenopausal osteoporosis comprehensively and systemically, thirty-two virgin Sprague-Dawley rats about 3-month-old were used and randomly divided into 4 groups: sham control group (Sham), OVX control group (OVX), naringenin treatment group and 17β-estradiol (E2) treatment group. After 12 weeks treatment with different drugs, 24 h urine were collected, organs were weighed and the organ indies were computed. Uterine pathological changes were observed by making paraffin section. Biochemical parameters and bone turnover markers: serum osteocalcin (BGP) and urine deoxypyridinoline (DPD) were analyzed with automatic biochemical analyzer or ELISA assay. Bone mineral density (BMD) and bone mineral content (BMC) were analyzed by DEXA, bone biomechanical properties was measured by three point bending test and the trabecular bone microarchitecture was evaluated by Micro CT. From the results, we can see that: the gaining of weight and the increasing of bone turnover markers such as serum BGP and urinary DPD could be inhibited by naringenin. The treatment could also enhance the bone strength and prevent the deterioration of trabecular microarchitecture, increase the bone volume, trabecular number and thickness, and decrease the trabecular space. The effects mentioned above were not accompanied with stimulating effects on uterus. Long-term using of naringenin had no obvious influence on other organs and the liver and kidney functions. The study suggests that naringenin had obvious antiosteoporotic effect on ovariectomized rats and it had the potential value for the treatment of postmenopausal osteoporosis.
Amino Acids ; urine ; Animals ; Bone Density ; Disease Models, Animal ; Estradiol ; pharmacology ; Estrogen Antagonists ; pharmacology ; Female ; Flavanones ; pharmacology ; Osteocalcin ; blood ; Osteoporosis ; drug therapy ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Uterus ; pathology

This paper described the results of the optimal method for preparation and extraction of lectin from Phaeoporus obliquus(Pers ex Fr).(J.Schroet).We determined the optimal buffer solution using the ag-glutination test with 2% hare blood cells.The optimal extraction process included the following parameters that were conformed by optimization analysis: material-water ratio,extraction time,concentration of sodium chloride and the pH value.The results are that the agglutination indexes of POL using TBS and PBS extrac-tion buffer were 64 and 16,respectively.The optimal extraction process is that,the material-water ratio was 1:50,the extraction time was 20 h,while the concentration of sodium chloride was 0 mol/L and pH was 8.0.The agglutinability of POL was 256 examined by this method.The optimized extraction process is stable and practicable,which may supply some basis for the application of the lectin from Phaeoporus obliquus in immunoregulation.

Objective To investigate the therapeutic effects of recombinant human endostatin (RHES) combined with radiotherapy on brain metastases (BM) of non-small cell lung cancer (NSCLC) and the patients suitable for this therapy.Methods Eighty patients with BM of NSCLC were randomly divided into RHES combined with radiotherapy group (combination group) and radiotherapy alone group (each group with 40 patients).The short-term effective rate,overall survival time,cerebral edema index and adverse reactions were observed and the expressions of vascular endothelial growth factor receptor 2 (VEGFR2) protein in primary lesions were detected with immunohistochemical method in all patients.Results Compared with radiotherapy alone group,brain edema was significantly relieved (t=4.9,P=0.000) and there were no marked adverse reactions in combination group.In short-term effective rate,there was no statistical significance in total population (n=80,90％ vs.75％,x2=3.11,P=0.07),but there was statistical significance in the patients with positive VEGFR2 (93％ vs.67.7％,x2=6.31,P=0.012).In overall survival time,there was no statistical significance in total population (n=80,P=0.35,95％ CI:0.25-1.30) or in the patients with positive VEGFR2 (P=0.109,95％ CI:0.40-1.34).Conclusion Compared with radiotherapy alone,RHES combined with radiotherapy can relieve brain edema in the patients with BM of NSCLC and obtain better short-term effective rate in the patients with positive VEGFR2.

Objective To report treatment of infectious bone and soft tissue defects caused by tibial shaft fracture of Gustilo type Ⅲ B using free flap and Ilizarov bone transport.Methods Nineteen patients who had suffered from infectious bone and soft tissue defects following tibial shaft fracture of Gustilo type Ⅲ11 B were treated from May 2010 to February 2015.They were 15 men and 4 women,aged from 21 to 58 years (average,45.3 years).Their course of disease ranged from 16 to 21 months,averaging 17.9 months.The area of their infectious defects ranged from 10 cm × 6 cm to 21 cm × 12 cm,and the length of their bone defects from 5 to 11 cm (average,7.4 cm).They were treated with debridement,simple external fixation to reconstruct bony support,coverage of wounds with free flap,and stuffing the dead space with antibiotic concrete beads,followed by Ilizarov bone transport and bone graft after control of infection to reconstruct the defective tibia and function of the affected limb.Results All the flaps survived.Necrosis occurred at the distal margin of one flap but responded to dressing.Pin tract infection occurred in 4 cases but also responded to dressing and antibiotic therapy.Autografts of iliac cancellous bone were implanted into the gliding and traction ends of the bone fragments one month after bone transport had come to rest.All the patients achieved direct bony union.The patients were followed up for an average of 25 months(range,from 19 to 36 months).No secondary fractures or angular deformity was observed.The total treatment time averaged 17.9 months,with no recurrence of infection.According to the Puno score system for functional evaluation at the last follow-up,7 cases were rated as excellent,6 as good and 6 as fair.Conclusion The infectious bone and soft tissue defects caused by tibial shaft fracture of Gustilo type Ⅲ B can be treated by free flap and Ilizarov bone transport,resulting in definitely positive outcomes.

Purpose To evaluate the application value of volume CT perfusion (CTP) protocol with 128-slice CT scanner in patients with unilateral middle cerebral artery (MCA) stenosis or occlusion.Materials and Methods Thirty-five patients with severe unilateral MCA stenosis or occlusion who underwent cerebral volume CTP examination in Wuhan No.1 Hospital between July 2013 and December 2014 were evaluated.The volume CTP parameter maps of cerebral blood flow (CBF),cerebral blood volume (CBV),mean transit time (MTT) and time to peak (TTP) were analyzed.Meanwhile,the dynamic CT angiography (4D-CTA) images were obtained.Results In three patients,CBF,CBV and MTT were observed normal but TTP was delayed.In the other thirty-two patients,CBF in the affected side was lower than that in the contralateral side (but difference without significance:P＞0.05).The increased CBV,prolonged TTP and MTT were also detected in the affected side compared with the contralateral side (difference with significance:P＜0.05).McNemar test showed 4D-CTA and DSA diagnosis of middle cerebral artery occlusion were not significantly different (P＞0.05),and they had better consistency (Kappa=0.861,P＜0.05).Conclusion Brain volume CTP protocol with 128-slice CT scanner provides valuable information about cerebral artery abnormalities and hemodynamic changes in patients with severe stenosis or occlusion of MCA.It has great value for clinical application.

Objective To discuss Bushen Huoxue Prescription mechanism of action on follicular granulosa cell apoptosis of premature ovarian failure (POF) mice. Methods The mouse zona pellucida 3 as the antigen, multi-point injection of subcutaneous immunization BALB/c to female mice was used to establish autoimmune POF model. Forty clean female mice were randomly divided into blank group, model group, Estradiol Valerate group and Bushen Huoxue Prescription group. All medicine groups were given relevant medicine for gavage, while model and blank groups were given normal saline with the same amount for gavage for 15 days. After the treatment, ovary, uterus, and thymus were weighed, and the viscera index was calculated. An ovarian slice was taken and HE staining was used to observe granulosa cell apoptosis. The levels of E2, LH, and FSH were detected by using the radioimmunoassay method. RT-PCR was used to analyze Fas and Fas-L mRNA expressions in ovarian granulosa cells. Results Compared with blank group, the weight of ovary, thymus, and uterus decreased significantly (P<0.05, P<0.01);Serum E2 level dropped (P<0.01);levels of FSH and LH, and expressions of Fas and Fas-L mRNA increased significantly (P<0.01). Compared with model group, granulosa cell apoptosis significantly decreased, and ovary index, uterus index, and thymus index increased (P<0.05);serum E2 level increased (P<0.01);levels of FSH and LH, and expressions of Fas and Fas-L mRNA significantly decreased (P<0.05). Conclusion Bushen Huoxue Prescription can decrease expressions of Fas and Fas-L mRNA to inhibit cell apoptosis by adjusting the hormone level of hypothalamus-pituitary-ovarianaxis.

Osteoclasts are multinucleated giant cell, which derived from mononuclear myeloid hematopoietic stem cells with the function of bone absorption. Osteoclasts plays a key role in bone metabolism, therefore the body is very strict to regulation of osteoclastogenesis. Mobilization and differentiation of osteoclast maturation is a complex and sophisticated multi-level regulatory processes. In the relevant regulatory mechanisms, OPG/RANKL/RANK system plays a pivotal role in the process of osteoclast differentiation and maturation. Recent studies revealed that immune cells and osteoclasts were closely connect with each other in the field of bone metabolism, also provide a new therapeutic target for the treatment of bone diseases. The apoptosis of osteoclasts in bone metabolism have been payed more attention,while its mechanism is still not clear, which need further research.
Animals ; Cell Differentiation ; Gene Expression Regulation ; Humans ; Osteoclasts ; cytology ; metabolism ; Osteoprotegerin ; genetics ; metabolism ; RANK Ligand ; metabolism ; Receptor Activator of Nuclear Factor-kappa B ; genetics ; metabolism