OBJECTIVE:To investigate the role of interferon to increase the sensibilization of MGMT positive glioma stem cel s to temozolomide in vitro. METHODS:Glioma cel lines, U251 and SKMG-4, were induced by suspended cloning bal formation method to harvest MGMT positive glioma stem cel s, U251G and SKMG-4G. Cel counting kit-8 assay was used to detect the kil ing effect of interferonα/βcombined with temozolomide on MGMT positive glioma stem cel s. RT-PCR and western blot assay were employed to determine the expression of MGMT and nuclear factorκB in MGMT positive glioma stem cel s. RESULTS AND CONCLUSION:Western blot results showed positive expression of MGMT in U251G and SKMG-4G cel s at protein levels. After intervention with interferonα/β, the mRNA expression of MGMT and nuclear factorκB in SKMG-4G and U251G cel s was reduced significantly, and then further decreased after temozolomide treatment. These findings indicate that interferonα/βcan remarkably strengthen the kil ing effect of temozolomide on MGMT positive glioma stem cel s.

Objective To analyze the clinical characteristics and the endocrine changes in children with craniopharyngioma,and to improve the pediatrician understanding of the disease.Methods The study subjects consisted of 14 children with craniopharyngioma admitted to the Department of Endocrinology,Beijing Children's Hospital Affiliated to Capital Medical University from Jan.2004 to Dec.2012.All the patients were followed up to analyze the clinical symptoms improvement,endocrine test results and medication,et al.Results The main clinical manifestations were headache (7/14 cases,50.0％),growth retardation(4/14 cases,28.6％),vomiting (4/14 cases,28.6％),polydipsia/ polyuria (3/14 cases,21.4％) and vision diminution (3/14 cases,21.4％).Three patients didn' t undergo the surgery,and 3 cases with diabetes insipidus and 2 cases with growth hormone deficiency,and 1 case with central hypothyroidism by laboratory test.The rest 11 children received surgery and all patients had changes in endocrine after it.Five cases got polydipsia and polyuria,other 5 cases had electrolyte disturbances,and 2 cases had epilepsy.Nine patients were followed up,and the follow-up duration ranged from 5 months to 10 years [(3.29 ± 3.52) years] after surgery.Seven patients got better and 2 patients got worse.Conclusions For clinical symptoms of increased intracranial pressure,changes in endocrine,the vision and visual field,the possibility of craniopharyngioma should be taken into account.Surgery is the main treatment,but it can lead to the damage of hypothalamus and pituitary gland.Changes in endocrine,electrolyte disturbances and epilepsy are the common complications.According to the level of endocrine,longterm hormone replacement therapy for some postoperative patients should be continued.

Objective To observe the effect of acute retrograde gastric electrical stimulation (RGES) on gastric accommodation,emptying and gastrointestinal hormones releasing in obese patients. Methods Sixteen obese patients were examined. On the first day,a pair of mucosal gastric electrodes was placed under endoscope. The liquid meal load test and the standard solid meal gastric emptying test were carried out on the second day. RGES was performed starting at 30 minutes before each test and through the whole testing process. The serum leptin,ghrelin,resistin and peptide YY were examined before and after the standard solid meal gastric emptying test. On the third day,sham stimulation was given. The effect of acute RGES on related index was compared by self-control.Results BMI of the 16 patients was (32. 90±2. 99) kg/m2. Acute RGES significantly reduced the liquid meal volume of fullness [(460±148) ml and (630±219) ml,t=-7. 200,P＜0. 01] and the maximal tolerable meal volume [(699±215) ml and (926±295) ml,t=- 5. 390,P＜0. 01]. The effects of RGES and sham RGES on half-emptying time of standard solid meal was (109±26) min and (103±31) min (t=1. 009,P= 0. 329);on the retention rate of standard solid meal at one hour and two hour was (63. 37±9. 75)% and (59. 73±12.87)% (t=1. 834,P= 0. 087),(42.22±13.97)%and (38. 33±16. 87)% (t= 1.780,P= 0. 095),respectively. The ratio of gastrointestinal hormones after and before the stimulation also of the sham stimulation,leptin was 1. 03±0. 34 and 1. 08±0. 38(t=-0.386,P=0. 705),ghrelin was 0. 99±0. 11 and 0. 98±0. 12 (t= 0. 413,P=0.685),resistin was 1. 11±0. 25 and 0. 99±0. 24 (t= 1. 753,P= 0. 100),and peptide YY was 1. 56±0. 71 and 1. 33±0. 61 (t=1. 402,P= 0. 181). Conclusions In obese patients,acute RGES significantly reduce the liquid meal volume by lower gastric accommodation,to certain extent which will delay gastric emptying. There is no significant influence on gastrointestinal hormones releasing.

Objective To evaluate the relationship between sternum protection and bone marrow suppression in postoperative radiotherapy for esophageal carcinoma. Methods Total of 98 postoperative patients with thoracic esophageal carcinoma were randomly divided into experimental group (52 cases) and control group (46 cases). All patients were given intensity-modulated radiotherapy (IMRT), with the dose of 50-50.4 Gy. The patients in experimental group were irradiated by 6 fields (4-fields in front, 2-fields behind) which were crossed to avoid direct exposure to the sternum. The patients in control group were irradiated by 5 fields (3-fields in front, 2-fields behind) with front-middle of the field passing through the sternum. Concurrently all patients received 2 cycles of cisplatin chemotherapy. Results Dmean, V20 and V30 of the sternum in the experimental group were (20.21 ±3.60) Gy, (40.78 ±7.19) % and (33.78 ±9.44) %, which were lower than those in the control group [(30.91±5.21) Gy, (81.01±4.81) %, (51.60±6.84) %], respectively (P<0.05). However, the volume and dose distribution of lung, spinal cord and heart were similar between the two groups (P> 0.05). Both the incidence rates of bone marrow suppression at 14th day and 35th day after radiotherapy were significantly higher in the control group (52.2%, 73.9%) than those in the experimental group (28.8 %, 50.0 %) (P< 0.05), and the incidence rate of bone marrow suppression at 7th day after radiotherapy was similar between the two groups. Conclusion Protecting and sketching for sternum in postoperative radiotherapy for esophageal carcinoma can reduce the incidence of bone marrow suppression effectively, which would not increase the radiation dose in the lung, heart and spinal cord.

Objective:To explore the effects of Bailing capsules combined with salvianolate on inflammatory cytokines, renal vas-cular endothelia and renal function in diabetic nephropathy inⅣphase. Methods:Totally 60 patients with diabetic nephropathy in IV phase were randomly divided into the control group (30 cases) and the treatment group (30 cases). The control group was given the conventional and salvianolate therapy, and the treatment group was treated with Bailing capsules additionally. The treatment course was 3 months. The changes of serum creatine(SCr), urean itrogen (BUN), creatinine clearance rate (Ccr), ethelin (ET-1), interleukin-6 (IL-6), tumor necrosis factor (TNF-a) and C-reactive protein (CRP) were detected before and after the treatment. Results:After the 3-month treatment, SCr, BUN, IL-6, TNF-a, CRP and ET-1 were significantly decreased in the two groups with statistical signifi-cance, and Ccr was significantly increased in the treatment group (P<0. 05), and the changes in the treatment group were significant-ly better than those in the control group (P<0. 05). After the treatment, the blood glucose, blood lipids, glycosylated hemoglobin and the other indicators had no significant changes in the two groups (P>0. 05). There was no significant difference in the incidence of adverse drug reactions between the two groups (P>0. 05). Conclusion:The combination of Bailing capsules and salvianolate can re-duce the inflammatory reaction and improve renal vascular endothelial function and renal function in the patients with diabetic nephrop-athy in Ⅳ phase with promising safety and few adverse reactions.

BACKGROUND:Establishment of in vitro culture system of human placental microvascular endothelial cel s with high purity is very important. In recent studies, some scholars have successful y obtained a large number of placental microvascular endothelial cel s by three-stepenzyme digestion and magnetic separation method, but the procedures were extremely complex and it had great damage to the cel s. Therefore, how to separate human placental microvascular endothelial cel s easily and obtain high-purified cel s has become a research hotspot.
OBJECTIVE:To investigate an efficient method to isolate and purify human placental microvascular endothelial cel s from early vil us microvessels, observe the cel growth and identify the cel s.
METHODS:The vil i from normal early pregnancies (6-8 weeks) after artificial abortion were col ected aseptical y. Using two-step digestion procedure and discontinuous Percol density gradient centrifugation method, human placental microvascular endothelial cel s were obtained. Then the cel s were identified by trypsin digestion method and repeated adherence method.
RESULTS AND CONCLUSION:Human placental microvascular endothelial cel s were isolated successful y from early vil i. The primary cel s adhered to the wal s after inoculated for 24 hours and entered logarithmic phase at 10 days. 80%of the cel s achieved a confluence at 12-13 days after inoculating. The subculture cel s grew swiftly with the typical cobblestone appearance. Immunofluorescence staining showed that, cultured human placental microvascular endothelial cel s demonstrated a strong positive reaction to von Wil ebrand factor antigen and CD31, accounting for 100%. MTT assay results showed that, human placental microvascular endothelial cel s at passage 5 exhibited an S-shaped growth curve. High-purity human placental microvascular endothelial cel s can be obtained by proteolytic enzymes digestion and discontinuous Percol density gradient centrifugation method, and the purity is detected by trypsin digestion method and repeated adherence method.

Objective To study the outcomes of selective reduction of triplet pregnancy by assisted reproductive technology. Methods The clinical data of 31 women who succeeded in conception by vitro fertilization-embryo transfer in the third affiliated hospital of Guangzhou Medical University were retrospectively investigated to analyze and compare the rates of abortion from triplet pregnancy, twin pregnancy and single pregnancy after selective reduction of triplet pregnancy, the incidence of pregnant complications, outcomes of perinatal period. Results There were no significant differences between triplet pregnancy and the twin and single pregnancy after selective reduction of triplet pregnancy in terms of pregnancy and parity time, fetal disease, premature rupture of membrane, severe eclampsism, gestational diabetes and postpartum hemorrhage (P>0.05). There were significant differences in gestational weeks, birth weight, rate of premature birth, rate of neonatal transfer to NICU and neonatal RDS: The gestational time in the triplet group longer than the groups of twin pregnancy and single pregnancy after selective reduction of triplet pregnancy (P<0.05). (37.3 ± 1.9) vs. (35.2 ± 0.9), (32.6 ± 2.3), respectively), the rate of premature birth dropped (100%vs. 100%, 33.3%, respectively), the body weight was increased (1 707 ± 360.4)g vs. (2 066.1 ± 307.5)g, (2 712.5 ± 514.1)g, respectively and the neonatal complication rate was reduced (P<0.05). Conclusion The selective reduction of multiple pregnancy may decrease the risk of premature birth and reduce the rate of lower body weight of neonates, improving the pregnancy outcomes, but the higher rate of abortion at the early or middle course of pregnancy after selective reduction is worth our attention.

Objective:To explore the effect of hydrogen sulfide (H 2S) on modulating the subunit Kir6.2 of adenosine triphosphate sensitive potassium channels via the cyclic guanosine monophosphate-dependent protein kinase (cGMP/PKG) signaling pathway in epileptic rat models. Methods:Sixty adult male SD rats were randomly divided into the following six groups (10 rats in each group) by random number table method: control, epileptic, H 2S donor, H 2S donor+epileptic, KT5823 (one inhibitor of the cyclic guanosine monophosphate-dependent protein kinase)+H 2S donor+epileptic, and glibenclamide (one inhibitor of the adenosine triphosphate sensitive potassium channels)+H 2S donor+epileptic groups. Except the control group, SD rats were intraperitoneally injected with plentylenetetrazole to make the kindling models and their behaviours were recorded including the latency period, the grade, and the duration of the first epileptic seizure according to the Racine′s standard. The waveforms of electroencephalogram (EEG) in hippocampus were also recorded during the seizure. The mRNA and protein levels of PKG and Kir6.2 in hippocampus were evaluated by Western blotting and quantitative real-time polymerase chain reaction, and the hippocampal concentrations of cGMP and phosphorylation of cyclic guanosine monophosphate-dependent protein kinase (p-PKG) were detected by enzyme linked immunosorbent assay. Results:Rats in the epileptic group showed Ⅳ-Ⅴ grade of epileptic seizure [4.500 (4.000, 4.875)], short latency period [(10.37±8.21) min] but long duration [(69.50±24.37) s] of seizure. Compared to the epileptic group, rats in the H 2S donor group showed Ⅱ-Ⅲ grade of epileptic seizure ( P=0.004), significantly longer latency period ( P<0.001), and shorter duration of seizure ( P<0.001). Compared to the H 2S donor+epileptic group, rats in the KT5823+H 2S donor+epileptic group showed Ⅲ-Ⅳ grade of epileptic seizures, significantly shorter latency period ( P<0.001), and longer duration of seizure ( P<0.001). The results of EEG showed that the wave patterns in the epileptic group were spike or sharp waves and the amplitudes were largest [(190.570±23.590) μV]. Compared with the epileptic group, amplitudes were reduced ( P<0.001) in the H 2S donor+epileptic group. PKG mRNA and PKG protein were expressed differently among all groups (PKG mRNA: n=5, H=26.714, P<0.001; PKG protein: n=5, F=30.597, P<0.001). Compared with the control group, the expression of both PKG mRNA and PKG protein was decreased (PKG mRNA: 1.000±0.001 vs 0.782±0.064, P=0.023; PKG protein: 0.550±0.037 vs 0.145±0.020, P=0.042) in the epileptic group. Besides, Kir6.2 mRNA and Kir6.2 protein were expressed differently among all groups (Kir6.2 mRNA: n=5, H=27.761, P<0.001; Kir6.2 protein: n=5, F=60.659, P<0.001). Compared with the control group, the expression of both Kir6.2 mRNA and Kir6.2 protein was decreased (Kir6.2 mRNA: 1.000±0.001 vs 0.897±0.033, P=0.004; Kir6.2 protein: 0.384±0.035 vs 0.215±0.016, P=0.024) in the epileptic group. And the concentrations of cGMP and p-PKG were decreased (cGMP: P<0.001; p-PKG: P<0.001) in the epileptic group. The results in the H 2S donor+epileptic group were up-regulated (PKG mRNA: P=0.047; PKG protein: P<0.001; Kir6.2 mRNA: P=0.011; Kir6.2 protein: P<0.001; cGMP: P<0.001; p-PKG: P<0.001) compared with the epileptic group. However, the results in the KT5823+H 2S donor+epileptic group were down-regulated (PKG mRNA: P=0.015; PKG protein: P=0.027; Kir6.2 mRNA: P=0.013; Kir6.2 protein: P=0.017; cGMP: P=0.005; p-PKG: P<0.001) compared with the H 2S donor+epileptic group. Conclusion:A possible mechanism is that H 2S prevents the epileptic seizure from modulating the subunit Kir6.2 of ATP sensitive potassium channels via the cGMP/PKG signaling pathway.

Objectives To explore the expression, biological function, and mechanism of MKI67 in pancreatic cancer and its clinical significance. Methods The expression level, diagnosis, and prognostic value of MKI67 in pancreatic cancer were analyzed using public databases. We also investigated the association between the MKI67 with immune cell infiltration and immune checkpoint molecules. We analyzed the functional pathway enrichment to uncover the possible molecular mechanisms. qRT-PCR and Western blot assay were used to verify the expression of MKI67 mRNA and protein. Immunohistochemistry staining was used to detect the expression of MKI67 in tissue protein. Results The high expression of MKI67 was significantly associated with high histological grades and poor outcomes in pancreatic cancer. High MKI67 expression was correlated with poor prognosis of pancreatic cancer patients (P=0.009). MKI67 was an independent risk factor for the patient outcome (95%CI: 1.084-1.743, P<0.05). The MKI67 expression was positively correlated with the helper T cell 2 levels but negatively correlated with plasmacytoid DC, NK cells, mast cells, the T follicular helper, immune DC, and CD8 T cells. Conclusion MKI67 may serve as a biomarker for the diagnosis and prognosis of pancreatic cancer and the mechanism might be associated with immune escape or immunosuppression.

Objective To investigate the effects of fall efficiency on activities of daily living for community elderly people so as to prevent them from falling.Methods A total of 169 community elderly residents in the city were investigated for the fall efficiency and activities of daily living using questionnaires. Results In the past one year, there were 32 people (18.9%) fall in the community environment with 41 falls (0.22 times per person per year);the differences of fall efficiency and activities of daily living between the fallers and non-fallers were statistically significant (P<0.05);a positive correlation was found between fall efficiency and activities of daily living in both fallers and non-fallers (P<0.05).Conclusions Fall rate in the investigated elderly people is high. Fall efficiency may not only contribute to the fall but also affect activities of daily living in community elderly people. The activities of daily living of elderly people can be reduced by the fear of falling.