BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.

Objective To investigate the imaging features of papillary tumor of the pineal region(PTPR).Methods The ima-ging data of 10 patients with PTPR confirmed by operation and pathology were analyzed retrospectively.Results All lesions were located in the posterior commissure area of the posterior inferior wall of the third ventricle.All lesions were heterogeneously hyper-intensity on T1WI and hyperintensity on T2WI.Multiple small cysts or microcapsules signal intensity were observed within the tumor.Of all 10 lesions,there were 9 lesions with high signal intensity within or at the edge of the lesion on T1WI.All lesions showed restricted diffusion.All 10 cases showed uneven and obvious enhancement patterns.Midbrain tectum was compressed and moved backward in 5 cases,moved downward in 2 cases,moved forward and downward in 1 case,and was not clearly displayed in other 2 cases.One case was disseminated and 1 case was hyperperfusion.There were all 10 cases with obstructive hydrocephalus and equal or slightly high density on CT imaging,and 4 cases with calcification.Conclusion Imaging characteristics of PTPR included the lesions centered on the posterior commissure,compressed tectum with backward and downward,multiple small cysts or micro-capsules components within the tumor,hyperintensity on T1WI,and uneven and obvious enhancement patterns.

Following the principles of evidence-based medicine,in accordance with the structure and drafting rules of standardized documents,based on literature research,according to the characteristics of chronic cough in children and issues that need to form a consensus,the TCM Guidelines for Diagnosis and Treatment of Chronic Cough in Children was formulated based on the Delphi method,expert discussion meetings,and public solicitation of opinions.The guideline includes scope of application,terms and definitions,eti-ology and diagnosis,auxiliary examination,treatment,prevention and care.The aim is to clarify the optimal treatment plan of Chinese medicine in the diagnosis and treatment of this disease,and to provide guidance for improving the clinical diagnosis and treatment of chronic cough in children with Chinese medicine.

Objective To investigate the MRI classifications and imaging findings of dysembryoplastic neuroepithelial tumor(DNET). Methods MR images of 34 patients with pathologic confirmed DNET of Beijing Sanbo Brain Hospital were retrospectively reviewed in this study. The classification was made according to the number of pseudocysts, scope of involvement, morphology and location. Results MRI appearances of DNET were divided into three subtypes: cystic‐like, polycystic‐like and diffuse type. Twelve cases had cystic cortical, including front lobe (5 cases), temporal lobe (5 cases), parietal lobe (2 cases). These cases presented quasi‐circular or oval shape, with hypointense on T1WI and strongly hyperintense on T2WI. T2‐FLAIR was observed hyperintense ring sign in the tumor periphery and the cystic content was close to CSF but having the largest difference to that of CSF, which signal was higher than CSF. Twenty cases were polycystic‐like, front lobe (7 cases), temporal lobe (7 cases), parietal lobe (5 cases), occipital lobe (1 case). In these 20 cases, they had slightly hypointense on T1WI and strongly hyperintense on T2WI. Located in the cortex and subcortical matter, with wedge shape, gyriform or triangle shape.On T2‐FLAIR, internal septation and hyperintense"ring sign"were observed. Two cases were diffuse type, bilateral (1 case), unilateral (1 case). In these 2 cases, diffuse lesions involving multiple areas with hyperintense ring and internal septation on FLAIR, including subcortical white matter, deep nucleus and periventricular area. Conclusions The MR appearances of DNET are variable. Understanding the MR imaging type of DNET might improve the MR diagnosis of DNET.

Objective To observe the effect of mild hypothermia on myocardial β-adrenergic receptor (β-AR) signal pathway after cardiopulmonary resuscitation (CPR) in pigs with cardiac arrest (CA) and explore the mechanism of myocardial protection. Methods Healthy male Landraces were collected for reproducing the CA-CPR model (after 8-minute untreated ventricular fibrillation, CPR was implemented). The animals were divided into two groups according to random number table (n = 8). In the mild hypothermia group, the blood temperature of the animals was induced to 33 ℃ and maintained for 6 hours within 20 minutes after return of spontaneous circulation (ROSC) by using a hypothermia therapeutic apparatus. In the control group, the body temperature of the animals was maintained at (38.0±0.5)℃ with cold and warm blankets. The heart rate (HR), mean arterial pressure (MAP), the maximum rate of increase or decrease in left rentricular pressure (+dp/dt max)were measured during the course of the experiment. The cardiac output (CO) was measured by heat dilution methods before CA (baseline), and 0.5, 1, 3, 6 hours after ROSC respectively, the venous blood was collected to detect the concentration of cTnI. Left ventricular ejection fraction (LVEF) was measured with cardiac ultrasound before CA and 6 hours after ROSC. Animals were sacrificed at 6 hours after ROSC and the myocardial tissue was harvested quickly, the mRNA expression of β1-AR in myocardium was detected by reverse transcription-polymerase chain reaction (RT-PCR), the contents of adenylate cyclase (AC) and cyclic adenosine monophosphate (cAMP) were detected by enzyme linked immunosorbent assay (ELISA), the protein content of G protein-coupled receptor kinase 2 (GRK2) was detected by Western Blot. Results After successful resuscitation, the HR of both groups were significantly higher than the baseline values, CO, ±dp/dt max were significantly decreased, MAP were not significantly changed, serum cTnI levels were significantly increased. Compared with the control group, HR at 0.5, 1, 3 hours after ROSC were significantly decreased in mild hypothermia group (bpm: 142.80±12.83 vs. 176.88±15.14, 115.80±11.48 vs. 147.88±18.53, 112.60±7.40 vs. 138.50±12.02, all 1 < 0.01), CO was significantly increased at 1 hours and 3 hours after ROSC (L/min: 3.97±0.40 vs. 3.02±0.32, 4.00±0.11 vs. 3.11±0.59, both 1 < 0.01), +dp/dt max at 3 hours and 6 hours was also significantly increased after ROSC [+dp/dt max (mmHg/s): 3 402.5±612.7 vs. 2 130.0±450.6, 3 857.5±510.4 vs. 2 562.5±633.9; -dp/dt max (mmHg/s): 2 935.0±753.2 vs. 1 732.5±513.6, 3 520.0±563.6 vs. 2 510.0±554.3, all 1 < 0.05], the cTnI was significantly decreased at 3 hours and 6 hours afher ROSC (μg/L: 1.39±0.40 vs. 3.24±0.78, 1.46±0.35 vs. 3.78±0.93, both 1 < 0.01). The left at 6 hours after ROSC in both groups was decreased as compared with that before CA. The LVEF in the mild hypothermia group was higher than that in the control group (0.52±0.04 vs. 0.40±0.05, 1 < 0.05). The mRNA expression of β1-AR, and concentrations of AC and cAMP in hypothermia group were significantly higher than those in control group [β1-AR mRNA (2-ΔΔCT): 1.18±0.39 vs. 0.55±0.17, AC (ng/L):197.0±10.5 vs. 162.0±6.3, cAMP (nmol/L): 1 310.58±48.82 vs. 891.25±64.95, all 1 < 0.05], GRK2 was lower than that in the control group (GRK2/GAPDH: 0.45±0.05 vs. 0.80±0.08, 1 < 0.05). Conclusion Mild hypothermia can reduce the degree of cardiac function injury after CPR, and its mechanism may be related to the reduction of impaired myocardial β-AR signaling after CPR.

Objective To investigate the significance of glomerular deposition of C4d in accessing the severity and prognosis of IgA nephropathy. Methods A total of 131 patients were recruited for the study. Immunofluorescence was used to detect the deposition of C4d in renal tissue of pa?tients with IgA nephropathy,and the relationship between C4d deposition and clinical and pathological parameters and renal remission was ana?lyzed. Results Totally 30 patients had glomerular deposition of C4d. Compared with the patients without C4d deposition,the patients with C4d deposition had significantly higher levels of serum creatinine,urinary protein excretion and C4d and higher prevalence of hypertension,but had sig?nificantly decreased levels of glomerular filtration rates. With the histopathological phenotypes segregated by Lee 's classification,the ratios of C4d deposition presented an increase(P=0.005). The patients with C4d deposition had more severe mesangial proliferation,endocapillary hypercellu?larity,segmental glomerulosclerosis and tubular?interstitial injury. The rates of renal remission were significantly lower in IgA nephropathy patients with C4d deposition than those without C4d deposition(P<0.001). Conclusion IgA nephropathy patients with C4d deposition have more se?vere clinical and pathological manifestations and lower rate of renal remission. Glomerular C4d deposition is expected to be an important pathologi?cal prognostic factor for predicting the prognosis of IgA nephropathy.

Objective To evaluate in vitro antifungal activities of five clinically common antifungal agents against Geotrichum silvicola,and to observe morphological changes of Geotrichum silvicola after treatment with terbinafine.Methods Five antifungal agents commonly used in clinic,including terbinafine (TER),amphotericin B (AmB),flucytosine (FC),fluconazol (FCZ) and itraconazole (ICZ),were tested in this study.According to the National Committee for Clinical Laboratory Standards (NCCLS) M27-A2 document and relevant literatures,a susceptibility test was performed to determine the minimal inhibitory concentration (MIC) values of the five antifungal agents against G eotrichum silvicola.Scanning electron microscopy (SEM) was used to observe morphological changes of Geotrichum silvicola treated with different concentrations of TER.Results The MIC values against Geotrichum silvicola were as follows:TER,0.01 μg/ml;AmB,0.4 μg/ml;FC,2 μg/ml;FCZ,2.69 μg/ml;ICZ,0.25 μg/ml.Geotrichum silvicola was sensitive to TER,AmB,FC and FCZ,and sensitive to ICZ in a dose-dependent manner.SEM revealed that after TER treatment,the surface of Geotrichum silvicola became rough and shrinking with irregular defects and holes,and even gave a fragment-like appearance.Conclusions Among the five tested antifungal agents,TER shows the strongest activity against Geotrichum silvicola.Within a certain range,the higher the concentration of TER,the more serious the damage to Geotrichum silvicola.

Objective To explore the correlations of the MRI findings and its pathological typing in the focal cortical dysplasia (FCD) .Methods MR images of 74 patients with FCD confirmed by operation and histopathologic examination were analysed retro‐spectively .MRI findings with FCD were divided into three subtypes including radial band type ,hyperintensity type and mild type . The correlation of the FCD MRI findings and pathological typing is analysed .Results In 74 patients with FCD ,there were radial band type in 12 cases ,hyperintensity type in 32 cases ,and mild type in 30 cases respectively .M RI finding of radial band type FCD showed a tail of increased T2WI/FLAIR signal tapering down to the lateral ventricle .Hyperintensity type FCD showed increased T2 WI/FLAIR signal in the cortex and subcortical white matter ,accompanied with focal cortical thickening .Mild type FCD showed T2 WI/FLAIR subtle hyperintense signal in cortex with or without focal cortical thickening ,but there was no hyperintense signal in subcortical white matter .Most of radial band type FCD were ⅡB in pathology .Most of hyperintensity FCD were ⅡA and ⅡB .Mild type FCD was more found to beⅠA orⅠB .Conclusion Analysing MRI features would improve the accurate diagnosis of FCD and help to infer the pathological type .

ObjectiveTo investigate the effect of recombinant interleukin-2 (rIL-2) combined with allicin in the treatment of pancreatic cancer and related mechanisms. MethodsA nude mouse xenograft model was established. A total of 60 nude mice were randomized into control group, rIL-2 treatment group, allicin treatment group, and combined treatment group. At 4 weeks after treatment, peripheral blood was collected, tumor volume, tumor weight, and survival time were recorded, and survival rates were calculated. Flow cytometry was used to analyze the apoptosis of tumor cells and measure the percentages of CD4+ T, CD8+ T, and natural killer (NK) cells, ELISA was used to measure the level of interferonγ (IFNγ), and Western blot was used to measure the expression of Bcl-2 protein in tumor tissue. An analysis of variance was used for comparison of continuous data between groups, and SNK-q test was used for further comparison between any two groups. ResultsAt 4 weeks after treatment, the rIL-2 treatment group, allicin treatment group, and combined treatment group showed significant reductions in tumor volume compared with the control group (all P＜0.05), and the combined treatment group showed the greatest reduction (P＜0.01). The combined treatment group had a tumor inhibition rate of 90.5% and a prolonged survival time (60% of the mice survived at 55 days). Compared with the control group, the combined treatment group showed significant increases in the apoptosis rate of tumor cells (233%±4.3% vs 90%±3.7%, P＜0.05) and the expression of pro-apoptotic protein Bcl-2. The treatment groups showed significant increases in the percentages of CD4+ T, CD8+ T, and NK cells compared with the control group (F=23.74, 26.38, and 1972, all P＜0.001). Compared with the other three groups, the combined treatment group showed a significant increase in the IFNγ level (F=9.84, P=0.026). ConclusionCombined treatment with allicin and rIL-2 can enhance the innate immunity and adaptive immunity and thus improve the survival of pancreatic cancer.

Objective To determine MR manifestations and pathologic types of benign meningiomas and their relationship with tumor recurrence.Methods There were 218 patients (160 females,58 males; age range 4-79 years) with benign meningiomas in the study,including 31 recurrent meningiomas (recurrence group)and 187 primary meningiomas (primary group).All patients were proved by postoperative pathology.Differences of pathological types and MRI manifestations between the recurrence group and the primary group were evaluated by using x2 test and rank sum test.Logistic regression analysis was performed by taking tumor recurrence as the dependent variable,and age,gender,vital structures involvement and pathologic types as independent variables.The recurrent time intervals were compared by rank sum test.Results There were 30 patients with intracranial vital structures involvement or extreintracranial communication tumors in the recurrent group,which was obviously higher than that of the primary group (61 patients).The difference was statistically significant (x2 =57.672,P =0.001).The tumors located in the skull-base and juxtasinus in the recurrent group were obviously more than those in the primary group,and difference was statistically significant (x2 =10.990,P =0.001).Multi-logistic regression analysis showed that the recurrent risk of benign meningiomas was elevated significantly only with vital structure involvement or extre-intracranial communication tumors (wald x2 =31.863,OR =3.820,P =0.001).The recurrent risk of dural sinus involvement was 3.820 times of cerebral artery trunk and cranial nerves involvement,and the risk of the latter was 3.820 times of the non-involved.There was no statistical difference between the two groups in pathology type,location,peritumoral edema,tumor morphology and tumor size.The relapse time of dural sinus involvement and cerebral artery trunk involvement in the recurrent group was 24(13 to 180) and 126(12 to 187) months,respectively.There was significant difference (Z =2.197,P =0.028).Conclusions It is more common that the recurrent benign meningiomas located in the skull base and juxtasinus.The recurrent risk significantly increases when benign meningiomas with vital intracranial structure involved or with extra-intracranial communication tumor.The relapse time of dural sinus involvement is possibly shorter than that of cerebral artery trunk involvement.MRI plays an important role in predicting tumor recurrence and prognosis of benign meningiomas.