Objective: To explore and obtain the isolation and culture methods of the highly purified primary rat brain microvascular endothelial cells (BMECs). Methods: The brains of 10 3-week-old Wistar rats were harvested by decapitation, removing the white matter and mincing into approximately 1 mm3. A highly purified brain microvascular fragments were obtained through enzymatic digestion twice, 20% bovine serum albumin (BSA) and 33% continuous Percoll density gradient centrifugation. They were incubated into gelatin-coated 35 mm dish plates and the endothelial medium containing 4 μg/ml puromycins was added. The medium was renewed after 2 days. The endothelial cell growth and its morphology were observed using an inverted microscope. The expression of BMECs markers factor VIII-related antigen (vWF) was detected by immunocytoehemical method and was identified. Results: The endothelial cells grew out around the adherent brain microvascular fragments after culturing for 24 hours, and the cells were spindle-shaped. A fusion was formed after 5 to 6 days. BMECs after the fusion showed a typical "cobblestone-like" appearance. Immunocytochemical analysis showed the vWF expression of BMECs was positive, while the glial fibrillary protein (GFAP) expression was negative. The purity of BMVECs reached more than 96%. Conclusion: This method may successfully obtain the highly purified rat primary BMECs.

Angiopoietins ; metabolism ; physiology ; Animals ; Cell Proliferation ; Endothelial Cells ; cytology ; physiology ; Humans ; Neoplasms ; blood supply ; Neovascularization, Pathologic ; physiopathology ; Neovascularization, Physiologic ; physiology ; Pericytes ; cytology ; metabolism ; physiology ; Proto-Oncogene Proteins c-sis ; metabolism ; physiology ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; physiology ; Receptor, TIE-2 ; metabolism ; physiology ; Signal Transduction

Study the serum level of HGF, Cys C and TGF-beta1 in type 2 diabetic nephropathy (DN), the infect of Pingshen decoction on those index. Selected 69 cases of 2 type DN and randomly divided into therapy group (36 cases) and control group (33 cases). The therapy group were treated with Pingshen decoction 1 dose/d, bid po. The control group were treated with NephritisShu tablet, 6 tablet, tid po. 8 weeks was a course. Before and after treatment, we examine the serum level of HGF, Cys C and TGF-beta1 by ELISA and immunonephelometry, and compare with 30 cases of healthy control group. The study demonstrates that before treatment, the serum level of HGF in both groups were significantly lower than healthy control group (P < 0.01), but Cys C, TGF-beta1 were significantly higher (P < 0.01). After treatment, the serum level of HGF of both groups were increased. The serum level of HGF of therapy group were significantly higher than of control group (P < 0.01), but the serum level of Cys C and TGF-beta1 were significantly lower than control group (P < 0.01). The serum level of HGF was correlated negatively with Cys C,TGF-beta1. In control group, the UAER, urine beta2-MG and quantity of 24-hour urine protein were significantly decreased after treatment (P < 0.01). The index of urine of therapy group were significantly lower than control group (P < 0.01). Results indicate that test of serum level of HGF and Cys C,TGF-beta1 of diabetic nephropathy have important clinical significance. Pingshen decoction can effectively intervene in the serum level of HGF and Cys C, TGF-beta1 and index of urine.
Aged ; Aged, 80 and over ; Case-Control Studies ; Cystatin C ; blood ; Diabetic Nephropathies ; blood ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hepatocyte Growth Factor ; blood ; Humans ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; blood

0.05),but the expression in controls were negative.The expression levels of PRAME gene at remission was decreased obviously,but increased again when the patients relapsed.Conclusions Expression of PRAME gene has a high level in childhood acute leukemia.The dynamic changes are closely related with the prognosis.It can be regarded as a candidate for detecting minimal residual disease in acute leukemia,and may have important implications for estimating the prognosis and guiding the chemical therapy.

Objective To evaluate dendritic cell vaccine immunotherapy in postoperative colorectal cancer patients. Methods 40 colorectal cancer patients were divided into two groups. Group A was not treated with the dendritic cell vaccine after chemotherapy, Group B was treated with four courses of autologous dendritic cell vaccine. The level of cytokines was tested before and after the therapy in both groups. DTH were tested after the last course of treatment. Results The levels of cytokines significantly increased in group B after vaccination compared with that in group B before vaccination and that in group A.8 patients were DTH positive in 15 patients that were tested after the vaccination. The PTS (progression-free survival) was 22 months in group B compared with 17 months in group A. The recurrence of the disease was not observed in patients with DTH positive. Conclusion DC vaccine in postoperative colorectal cancer patients improves the immune status and elicits tumor-specific response.

Objective:This study aimed to compare rectal cancer tumor volume parameters measured by MRI sequences (T1WI, T2WI, and DWI) and/or CT with those by pathological specimen. Methods:Twenty-two patients with rectal cancer were prospectively enrolled. MRI sequences including T1WI, T2WI, and DWI, and/or CT of the pelvis were performed before operation. Volume parameters, such as tumor length along the rectal axis, maximum tumor width perpendicular to rectal axis, and tumor actual area in that perpendicular plane, were measured on T1WI, T2WI, DWI, and CT, respectively, for each patient. The respective pathological parameters were further measured in surgical specimen after total mesorectal excision. The two kinds of parameter values measured in imaging and pathology were statistically compared and accuracy appraisal was performed. Results:The mean Lpath-L was 4.06±1.14 cm. The mean LT1-L, LT2-L, LDWI-L, and LCT-L were 3.91± 1.51, 4.62±1.41, 3.39±1.05, and 3.94±1.23 cm, respectively. Correlation coefficients were 0.688, 0.635, 0.688, and 0.720 (P<0.05). An average 6 mm overestimation was found in T2WI, and 1 to 6 mm underestimation in T1WI, DWI, and CT in length values compared with those measured in surgical specimen. The mean Lpath-W was 2.56 ±0.94 cm. The mean LT1-W, LT2-W, LDWI-W, and LCT-W were 3.62±0.99, 3.66±0.76, 3.23±0.58, and 3.64±1.04 cm, respectively. The magnitude of mean overestimation ranged from 5.1 to 11.1 mm. The Apath was 4.30 ±2.83 cm2. AT1, AT2, ADWI, and ACT were 8.98±3.90, 8.99±3.43, 8.41±3.09, and 9.63±4.40 cm2, respectively, which double overestimated the tumor area in the perpendicular rectal plane. Conclusion:The difference in longitudinal length between MRI sequences/CT and pathological specimen was in the range of?6 mm to 6 mm. The mean maximum tumor width and areas in the maximum tumor perpendicular plane were overestimated. This study indicated that gross tumor volume delineation based on CT or MRI for rectal cancer irradiation should be conservative in the axial images of rectum, and meticulous consideration is required along the rectum.

Objective: To understand the effect of dobutamine on the integrated backscatter of myocardium. Methods: Nineteen open-chest dogs were detected by HP Sonos 5500 with S8 transducer, and the WT, AIBS and CVIB were measured before and after the dobutamine stress. Results: During the dobutamine stress, following the increase of WT, the transmural, subepicardial and subendocardial CVIB all increased. And the increase of subepicardial CVIB was greater than that of subendocardial CVIB, leading to the decrease of TGI. But the AIBS didn't change significantly in the whole process. Conclusion: Dobutamine can enhance myocardium CVIB but has no obvious infleunce on AIBS.

AIM: Combine disproportionality analysis with dynamically interactive graphics to understand spontaneously-reported adverse events in pharmacovigilance. METHODS: Four statistical methods, including Reporting Odds Ratio, Proportional Reporting Ratio, Multi-Item Gamma Poisson Shrinker and Bayesian Confidence Propagation Neural Network that are used for computing disproportionality are described. Tree maps and other graphical techniques are used to display the disproportionality results. RESULTS: Spontaneously-reported adverse events in pharmacovigilance are collected from physicians, patients, or the medical literature by regulatory agencies, pharmaceutical companies and device manufacturers to monitor the safety of a product once it reaches the market. In order to identify potential safety-signals, disproportionality analysis methods compare the rate at which a particular event of interest co-occurs with a given drug with the rate this event occurs without the drug in the event database. Tree maps are employed to interactively display the adverse events for particular drugs and compare the adverse events among the drugs. CONCLUSION Interactive graphical displays of disproportionality allow the analyst to quickly identify safety signals and perform additional follow-up analyses. Combining statistical methods with dynamically interactive graphics affords insights into the data inaccessible by traditional analysis methods.
Adverse Drug Reaction Reporting Systems ; statistics & numerical data ; Bayes Theorem ; Data Interpretation, Statistical ; Databases, Factual ; Drug-Related Side Effects and Adverse Reactions ; Humans ; Odds Ratio ; Pharmacovigilance ; Product Surveillance, Postmarketing ; statistics & numerical data

Background:Q fever endocarditis,a chronic illness caused by Coxiella burnetii,can be fatal if misdiagnosed or left untreated.Despite a relatively high positive rate of Q fever serology in healthy individuals in the mainland of China,very few cases of Q fever endocarditis have been reported.This study summarized cases of Q fever endocarditis among blood culture negative endocarditis (BCNE) patients and discussed factors attributing to the low diagnostic rate.Methods:We identified confirmed cases of Q fever endocarditis among 637 consecutive patients with infective endocarditis (IE) in the Peking Union Medical College Hospital between 2006 and 2016.The clinical findings for each confirmed case were recorded.BCNE patients were also examined and each BCNE patient's Q fever risk factors were identified.The risk factors and presence of Q fever serologic testing between BCNE patients suspected and unsuspected of Q fever were compared using the Chi-squared or Chi-squared with Yates' correction for continuity.Results:Among the IE patients examined,there were 147 BCNE patients,of whom only 11 patients (7.5％) were suspected of Q fever and undergone serological testing for C.burnetii.Six out of 11 suspected cases were diagnosed as Q fever endocarditis.For the remaining136 BCNE patients,none of them was suspected of Q fever nor underwent relevant testing.Risk factors for Q fever endocarditis were comparable between suspected and unsuspected patients,with the most common risk factors being valvulopathy in both groups.However,significantly more patients had consulted the Infectious Diseases Division and undergone comprehensive diagnostic tests in the suspected group than the unsuspected group (100％ vs.63％,P =0.03).Conclusions:Q fever endocarditis is a serious yet treatable condition.Lacking awareness of the disease may prevent BCNE patients from being identified,despite having Q fever risk factors.Increasing awareness and guideline adherence are crucial in avoiding misdiagnosing and missed diagnosing of the disease.

OBJECTIVETo observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α) in IL-1β induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages.

METHODSNaringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1β-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1β-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1β and TNF-α in IL-1β-induced CDs was detected by RT-PCR.

RESULTSThe appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1β on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α in IL-1β induced CDs (P < 0.05), which was approximate to the level of the normal group.

CONCLUSIONSNaringin could not only promote the proliferation of CDs, but also protect IL-1β-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1β and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

Animals ; Blotting, Western ; Cartilage, Articular ; Caveolins ; Chondrocytes ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; pharmacology ; Interleukin-1beta ; metabolism ; Polypodiaceae ; Rabbits ; Rhizome ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism