72 cases of treatments on insomnia were recorded in the medical books of Ming and Qing dynasties(A.D.1368～1911 years).The compatibility regularity of treating insomnia in these 72 cases were statistically analyzed with frequency and the R type cluster analysis methods. Combining the data and modern recognition, compatibility and medication regularity of treating insomnia was further analyzed.

Objective To observe the effects of propylene glycol mannate sulfate(PGMS) on the expression of ICAM-1 and VCAM-1 in kidney of diabetic rats, and explore the protective effect of PGMS on kidney. Methods Diabetes was induced by injecting STZ. Then the rats were randomly divided into four groups: normal control group, diabetic modal group, treatment group with high-dose and low-dose of PGMS. 8 weeks later, the blood glucose concentrations, HbA1c, the ratio of kidney weight to body weight, urinary albumin excretion rate (UAE ) in 24 hours were examined. The ICAM-1 and VCAM-1 located were detected by immunohistochemistry, while the expression of ICAM-1 and VCAM-1 were detected by Western blot method and the levels of ICAM-1 and VCAM-1 mRNA by RT-PCR.Results PGMS could reduce kidney hypertrophy and lower urinary albumin excretion rate (UAE)of 24-hour significantly. Immunohistochemical study with light microscopy demonstrated that the levels, both ICAM-1 and VCAM-1 in the kidney of test groups decreased obviously. PGMS down-regulated the mRNA and protein levels of ICAM-1 and VCAM-1 significantly . A positive correlation between expression of ICAM-1,VCAM-1 and kidney lesion was observed. Conclusion: PGMS decreased the expression of ICAM-1 and VCAM-1 in diabetic rats.

Objective To investigate the correlation between pathologic features and contrast?enhanced ultrasound in patients with breast ductal papilloma. Methods From January 2014 to January 2016,80 patients with breast intraductal tumor treated in our hospital were selected,including 20 cases of breast ductal carcinoma (ductal carcinoma group) and 60 cases of benign tumor (benign group). Both groups received conventional ultrasound, ultrasonic angiography,and pathological feature analysis. Results Breast surrounding hyperecho,internal micro calcification,border edge burr and posterior echo attenuation were more frequently seen onconventional ultrasound in ductal carcinoma groupthan inbenign group(P<0.05). The average scores of contrast?enhanced ultrasound were significantly higher in ductal carcinoma group than in the control group(P<0.05). In the ductal carcinoma group, there were statistical significant differences betweenpatientswith different lymph node metastasis dukes staging ,and differentiation types (P < 0.05). Spearman rank correlation coefficient analysis showed that breast intraductal carcinoma lymph node metastasis and dukes staging,differentiation types were markedly correlated with pathological features and ultrasound imaging (r=0.341,0.368,and-0.289;P<0.05). Conclusions Conventional ultrasound and contrast?enhanced ultrasound have very good imaging featuresin the diagnosis of breast intraductal cancer ,and there is correlation between pathologic features and contrast?enhanced ultrasound in patients with breast ductal papilloma,which can provide values for early diagnosis of breast ductal carcinoma.

This study is to investigate the inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide(LPS)-stimulated HMC-1 mast cells. The cytotoxicity of kaempferol to HMC-1 mast cells were analyzed by using MTT assay and then the administration concentrations of kaempferol were established. Histamine, IL-6, IL-8, IL-1β and TNF-α were measured using ELISA assay in activated HMC-1 mast cells after incubation with various concentrations of kaempferol (10, 20 and 40 µmol.L-1). Western blot was used to test the protein expression of p-IKKβ, IκBα, p-IκBα and nucleus NF-κB of LPS-induced HMC-1 mast cells after incubation with different concentrations of kaempferol. The optimal concentrations of kaempferol were defined as the range from 5 µmol.L-1 to 40 µmol.L-1. Kaempferol significantly decreased the release of histamine, IL-6, IL-8, IL-1β and TNF-α of activated HMC-1 mast cells (P<0.01). After incubation with kaempferol, the protein expression of p-IKKβ, p-IKBa and nucleus NF-κB (p65) markedly reduced in LPS-stimulated HMC-1 mast cells (P<0.01). Taken together, we concluded that kaempferol markedly inhibit mast cell-mediated inflammatory response. At the same time, kaempferol can inhibit the activation of IKKβ, block the phosphorylation of IκBα, prevent NF-KB entering into the nucleus, and then decrease the release of inflammatory mediators.

AIM The proofs of evidence-based medicine of Qingkailing Injection's effect on viral hepatitis are lack in spite of the wide use of Qingkailing Injection in medical practice.This study aims to use Meta-analysis to evaluate its clinical effect and safety.METHODS Such databases as CNKI,VIP and Wanfang were searched for randomized controlled trials related to Qingkailing Injection in treatment of viral hepatitis,which were published officially between 1996--2016.Revman 5.3 software was used to conduct Meta-analysis for the effective rate,rate of adverse reaction,the decrease rate of ALT,AST and TBIL of Qingkailing Injection.Meanwhile,subgroup analysis was conducted to the clinical effect of doses [＜ 0.5 mL/(kg · d),0.5-0.67 mL/(kg · d),＞0.67 mL/(kg · d)],treatment courses (≤ 14 d,＞ 14 d),manufacturers (Beijing University of Chinese Medicine Pharmaceutical Factory,Shenwei Pharmaceutical Company,Shanxi Taihang Pharmaceutical Company),year (1996 to 2002,2003 to 2009,2010 to 2016).RESULTS Fourteen studies were included according to the criterion.The results of Meta-analysis showed that the total effective rate,the decrease rate of ALT,AST and TBIL in Qingkailing Injection group were significantly higher than those in the control group (P ＜ 0.05).Whereas,no significiant differences were found between Qingkailing Injection group and the control group of its rate of adverse reaction (P ＞ 0.05).Subgroup-analysis showed that the effective rate of Qingkailing in doses subgroup,treatment courses subgroup,manufacturers subgroup and year subgroup were significantly higher than those of the control group.CONCLUSION Qingkailing Injection has obvious benefit in the treatment of viral hepatitis,which can reduce the activity of ALT and AST,also can reduce the content of TBIL.But we should pay attention to the prevention of allergic reaction in the clinical use.

Objective To investigate the anti-tumor immune response induced by human pancreatic cancer mucin 4 mRNA and human telomerase reverse transcriptase (hTERT) mRNA cotransfected dendritic cells (DC),and to provide the experimental evidences for the treatment of pancreatic cancer with multi-epitope loaded DC vaccine.Methods DC were isolated from peripheral blood mononuclear cells (PBMC) of six patients with HLA-A2+ pancreatic cancer and cultured.Mucin 4 mRNA and hTERT mRNA were transcripted and amplified in vitro,which were transfected into DC separately or in order by eleetroporation.DC were cultured for 48 hours.The expressions of mucin 4 and hTERT in DC were detected by quantitative real-time polymerase chain reaction (PCR) and Western blot.The survival rates of transfected DC were determined by methylthiazolyl tetrazolium (MTT) method.The cytotoxic T lymphocyte (CTL) activation induced by mucin 4 mRNA and hTERT mRNA transfected DC were evaluated by interferon (IFN)-γ release assays (enzyme linked immunosorbent assay) method.The cytotoxicity of CTL induced by mucin 4 mRNA and hTERT mRNA transfected DC in pancreatic cancer cell lines MiaPaCa-2,Capan-2,AsPC-1 and Pane-1 was measured by 51Cr standard cytotoxicity test.Student t test was performed for statistical analysis.Results After in order transfection of mucin 4 mRNA and hTERT mRNA for 48 h,the relative quantity of the expression of mucin 4 and hTERT in DC were 30.09±5.24 and 12.87±3.36,which were lower than the relative quantity of the expression in DC transfected separately (38.54±6.21 and 36.35±5.03,t=3.469,6.721,both P＜0.05).After transfected in order for 96 hours,the survival rate of DC decreased to 52.17％,which was lower than that of DC transfected separately (around 80％).The quantity of IFN-γ releasing of specific CTL induced by mucin 4 mRNA and hTERT mRNA cotransfected DC was (32.57±2.01) U/mL in 24 hours,which was higher than that of CTL induced by DC transfected with mucin 4 mRNA ((23.06±4.74) U/mL) or hTERT mRNA ((16.82±3.67) U/mL) separately (1=5.092,7.141,both P＜0.05).After co-transfected with mucin4 mRNA and hTERT mRNA,DC could effectivly induce HLA-A2+/mucin 4+/hTERT+ specific CTL immune responses,however there was no significant cytotoxicity in HLA-A2+ pancreatic cancer cells.Conclusion The induction of CTL anti-tumor immune response by DC co-transfected with mucin4 mRNA and hTERT mRNA is more significant compared with that by single antigen loaded DC.

Objective To investigate the apoptosis inducing effects of oridonin on leukemic THP-1 cells and its mechanisms of action. Methods THP-1 cells in culture medium in vitro were given different concentrations of oridonin (16～56 μmol/L) for 24, 48 and 72 h. The inhibitory rate of the cells were measured by MTT assay, apoptotic morphology was observed by Hoechst 33258 staining, and Annexin V/PI staining was used to detect cell apoptosis by flow cytometry (FCM). Caspase-3 and poly (ADP-ribose) polymerase (PARP) expression were detected by Western blotting. Results Oridonin (over 32 μmol/L) could inhibit the growth of THP-1 cells and cause apoptosis remarkably, the suppression was both in time-and dose-dependentmanner. Marked morphological changes of cell apoptosis such as condensation of chromatin was clearly observed by Hoechst 33258 staining, and Annexin V/PI staining showed that apoptotic cells gradually increased after the cells treated with oridinon. Western blotting showed cleavage of the caspase-3 zymogen protein (32×103), with the appearance of its 20×103 subunit, and a cleaved 89×103 fragment of 116×103 PARP was also found. Conclusion Oridonin can inhibit cell growth and induce apoptosis in THP-1 cells via activation of caspase-3. The results indicated that oridonin might be an important potential anti-leukemia reagent.

Objective To explore the impact of hypoxia/reoxygenation stimulation on phenotype and immune activity of dendritic cells(DCs) cultured from murine bone marrow. Methods Mouse DCs were generated from bone marrow cells and were divided into control group and hypoxia/reoxygenation group. DC in control group was cultured at normal condition, and in hypoxia/reoxygenation group was cultured at hypoxic condition for 4 h followed by cultured at normal condition for 24 h. Flow cytometry and mixed lym-phocyte reaction(MLR) was used to detect the phenotype and functional properties of DCs. ELISA was used to detect the concentration of TNF-α, IFN-γ and IL-12 in the supernalant, Imrounochemistry was used to de-tect the concentration of NF-κB. Results Hypoxia/reoxygen stimulation increased the CD80, CD86, MHC Ⅱ in the cytomembrane of DCs and TNF-α, IFN-γ, IL-12 concentration in the supernalant. Hypoxia/reoxy-gen stimulation also promoted the shift of NF-κB to karyon. Conclusion Under hypoxia/reoxygen stimula-tion, DCs express high level of surface molecules, and possess strong immune activity.

Objective To investigate the apoptosis inducing effect of peroxisome proliferator-activated receptor r(PPARr)agomst on leukemic K562 cells and its mechanisms of action.Methods K562 cells in culture medium in vitro were given different concentrations of PPARragonist rosiglitazone(RGZ)(20-80 umol/L)for O,24,48 and 72h The inhibitory rate of the cells were measured by MTT assay,cell apoptosis was detected by Annexin V/PI staining,and the expression of P53 protein as well as the activity of caspase-3 were also detected.Results RGZ(over 40umoL/L)could inhibit the growth of K562 cells and cause apopto-sis remarkably,the suppression Was both in time-and dose-dependent manner.The expression of P53 pmtein was upregulated and the activity of caspase-3 Was increased concomitantly after the cells werle treated by RGZ.Conclusion PPARr agonist RGZ(over 40 umol/L)can induce apoptosis on K562 cells signifieantly,upregu-lation the expression of P53 protein as well as increasing caspase-3 activity may be one of its most important mechamisms.

Objective To evaluate the relationship between stenosis of intra- or extra-cranial cerebral large artery and capsular warning syndrome(CWS). Methods Eleven consecutive CWS patients hospitalized during period of time from November 2008 to December 2009 were retrospectively analyzed.Result In these 11 patients with CWS, 5 patients had motor symptoms only, 4 patients had pure sensory symptoms, and 2 patients had sensorimotor symptoms. Ten patients underwent cervical contrast-enhanced MRA and intracranial MRA examination. The results showed no sign of arterial stenosis. Seven CWS patients eventually had strokes, 1 progressed to stroke despite receiving the therapy of antiplatelet and anticoagulation. All stroke lesions were located in the capsula interna. All the CWS patients had vascular risks: 7 were smoker, 8 had hypertension, 1 had diabetes mellitus, and 5 had hyperlipidemia. One patient had a history of previous stroke; no patient had a history of ischemic heart disease or atrial fibrillation. At follow-up(10. 2 ±3.4)mouths, the average modified Rankin scale score for all patients was 0. 73 ± 1.20.Conclusion CWS was not associated with stenosis of the intra and extra-cranial large cerebral arteries.CWS may be associated with small-vessel single-penetrator disease.