Background and purpose:Protein kinase C (PKC) is a potentially important target for cancer therapy due to its potential role in carcinogenesis. Abnormal expression and increasing activity of PKC-?are present in non-small cell lung cancer (NSCLC).The aim of this study is to investigate PKC-? protein expressions in lung cancer tissues by tissue microarray and their significance. Methods:Using tissue microarray, PKC-? protein expression in lung cancer patients was examined by immunohistochemistry. 182 samples of umor tissues and 18 samples of adjacent tissues were taken;of the 182 samples, 160 were NSCLC tissues and 22 were SCLC;102 samples from stage I cases and 58 from stageⅡ-Ⅲ cases.Results:Expression of PKC-? protein was observed in 48.3% (88/182) of cancerous tissues and 16.7% (3/18) of adjacent tissues (P

Objective To investigate the effect of berberine on gut microbiota and T helper cell 17 (Th17), regulatory T cell (Treg) cell in sleep deprived rats.Methods SD rats were randomized into control group, model group, low-dose and high-dose group (BBR1 and BBR2, 100 mg/kg and 200 mg/kg, administrated orally).Sleep deprived rat model was established by the small-platforms-over-water method.The number of bacteria in rectum content of rat was detected.The ratio of Th17/Treg was evaluated by flow cytometry.The expression of interleukin 17 (IL-17), RAR-related orphan receptor (ROR) C, and Forkhead Box Protein P3 (Foxp3) mRNA was evaluated.Results The counting of Clostridium perfringens was elevated (P<0.05), the amount of other microbiota decreased (P<0.05), Th17/Treg ratio(P<0.05), IL-17 and RORC expression enhanced (P<0.05), Foxp3 expression decreased (P<0.05) in the gut of model rats.In contrast, treatment with berberine inhibited the proliferation of Clostridium perfringens(P<0.05), and promoted the growth of other microbiota (P<0.05), and dampened Th17/Treg ratio (P<0.05), regressed IL-17 and RORC expression, augmented Foxp3 expression.Conclusions Various doses of berberine can counteract gut microbiota imbalance and Th17/Treg imbalance induced by sleep deprivation.

Objective To discuss the advantage of hispectral index(BIS) monitoring used in gastrointestinal endoscopy,and to observe elinical effects of target-controlled infusion(TCI) propofol combined with remifentanyl in elderly patients.Methods 120 patients aged from 65 to 78 with ASA physical status Ⅰ ～ Ⅱ were randomly divided into two groups,i,e.Group A (propofol with fentanyl ),Group B ( TCI propofol and remifentanyl with BIS).The amount of propofol was adjusted by consciousness and hemodynamics in Group A,while by BIS value in Group B.BIS was controlled at 55 ～60.The total amount of propofol and the time of wake-up,the incidence of complicatiors in each group were recorded.Results Compared with Group A,the changes of MAP and HR at T1 and T2 were smaller in Group B(P ＜ 0.05 ) ; Compared with Group A,the total amount of propofol and the time of wake-up were less in Group B ( P ＜ 0.05 ) ;Compared with Group A,the incidence of intraoperative respiratory depression and bradycardia were increased in Group B( P ＜ 0.05 ),there were no differences in the rest between the two groups.Conclusion Under BIS monitoring,TCl propofol and remifentanyl in elderly patients undewent gastrointestinal endoscopy may reduce propofol,and reach rapid recovery from anesthesia,but it needs additional management of respiration and circulation.

ObjectiveTo observe the changes of HR,MAP,BIS and Narcotrend induced by different doses of propofol in elderly patients. MethodsOne hundred elder patients(60 ～ 85 years old), AS A class Ⅰ or Ⅱ, scheduled for selective surgeries,were divided equally into 5 different doses of propofol( constant intravenous injection for 1min) groups of 0.5mg/kg( Ⅰ ) ,0. 75mg/kg( Ⅱ ) ,1.0mg/kg(Ⅲ) ,1.25mg/kg( Ⅳ)and 1.5mg/kg(Ⅴ). HR,MAP,BIS and Narcotrend were monitored before propofol injection and at 1 and 5 min after propofol injection. ResultsHR of 5 group s as similar. At 1 min after pmpofol injection, MAP decreased remarkably compared with at before in all 5 groups( t =2. 17,2.84,2.49,5.63,7.10, all p ＜ 0.05 ), which in group Ⅳ and Ⅴ decreased significantly compared with at in group Ⅰ, Ⅱ and Ⅲ(t =4.67,2.77,2.45,5.49,4.57,2. 18,all P＜0.05).At 1 min after propofol injection,BIS and NI values decreased compared with at before in all 5 groups(t =7.74,11.74,28.18,30.34,45.28, 6. 65,10.52,17.27,26.28,30. 14,allP ＜0.05) ,which in groupⅢ, Ⅳ and Ⅴ were significantly lower than those in group Ⅰ and Ⅱ (t =12.59,11.08,16.72,15.12,17.67,15.64,allP＜0.05).Dose of propofol was negatively correlated with BIS and NI value ( r =-0. 898/0. 930, P ＜ 0.01 ). ConclusionPropofol 1.0mg/kg constant injection should meet the sedation and hypnosis demand of general anesthesia in elderly patients and could not inhibit circulatory system; Bispectral Index and Narcotrend could accuratly monitor depth of anesthesia in elderly patients.

Objective: To establish and optimize the HPLC fingerprints of Angelica sinensis medicinal material and determine the ligustilide content to improve the quality standard for Angelica sinensis and improve the quality control level of Chinese angelica medici-nal material and preparations. Methods: A method for the determination of ligustilide was optimized by HPLC. The column was eluted on an Agilent ZORBAX SB C18(250 mm×4. 6 mm, 5 μm) column with acetonitrile-water (60 ∶ 40) as the mobile phase. The flow rate was 1. 0 ml·min-1, the detection wavelength was 326 nm, and the column temperature was at 35 ℃. The HPLC fingerprints of 13 batches of Angelica sinensis from different origins and methodological investigations were established and validated to set up an HPLC fingerprinting evaluation method for Angelica sinensis. Acetonitrile-0. 1% formic acid was used as the mobile phase with gradient elu-tion. The flow rate was 1. 0 ml·min-1, the detection wavelength was 280nm, and the column temperature was at 25℃. Results: The results showed that under the above HPLC conditions, ligustilide had good linearity within the range of 0. 032 3-0. 645 5 mg·ml-1(r=0. 999 9), and the average recovery was 100. 5% (RSD=1. 61% ,n=6). The quality fraction of ligustilide in Angelica sinensis was 0. 885 6%-2. 382 2% . Through the establishment of HPLC fingerprints of Angelica sinensis, the characteristic profiles with better peak shape and degree of separation and 18 common peaks with better resolution were obtained. The similarities of the 13 batches of angelica were all between 0. 9 and 1. 0. Conclusion: According to the methodological investigation, the HPLC fingerprints and ligustilide con-tent determination method of Angelica sinensis are simple, reliable, stable and feasible.

OBJECTIVE： To provide reference for improving the quality standard of Pheretima. METHODS： The contents of hypoxanthine and inosine in medicinal material samples were determined by HPLC. HPLC fingerprint of Pheretima was established according to “Similarity evaluation system for TCM chromatogramtic fingerprint” （2012 edition） software， and similarity evaluation was conducted. The determination was performed on Purospher STAR RP-18 endcapped with mobile phase consisted of methanol-water （gradient elution） at the flow rate of 1 mL/min. The detection wavelength was 248 nm， and the column temperature was set at 30 ℃. The sample size was 20 μL. RESULTS： The results of methodological investigation of content determination showed that the linear range of hypoxanthine and inosine were 1.58-31.6 μg/mL （r＝0.999 9）， 5.52-110.4 μg/mL（r＝0.999 8）， respectively. limits of quantify were 0.316， 0.552 μg/mL， respectively； limits of detection were 0.158， 0.110 μg/mL， respectively； RSDs of precision， stability （24 h） and repeatability tests were all less than 2.0％ （n＝6）. Average recovery rates were 103.0％ （RSD＝1.7％， n＝6） and 101.2％ （RSD＝1.2％， n＝6）， respectively. HPLC fingerprint for 15 batches of samples were established， and 8 common peaks were identified. The similarity of HPLC fingerprint of 14 batches of sample with control fingerprint R was higher than 0.900. CONCLUSIONS： The established method for content determination of hypoxanthine and inosine and HPLC fingerprint of Pheretima are simple， accurate and reproducible， and can be used for quality control of Pheretima.

Objective : To explore the relationship between the methylation level of RABL6 in peripheral blood and early lung cancer (LC) with a case-control study in the Chinese population． Methods : The methylation levels of 7 CpG sites in RABL6 gene in peripheral blood of samples from 275 LC patients (81．5% at stage I) ，and age- and gender-matched 185 benign lung nodule cases and 267 matched healthy controls were measured by matrix-assisted laser desorption ionization time-of-flight mass spectrometry．Multinomial Logistic regression adjusted for covariates was used to analyze the association between the RABL6 methylation and LC．Mann-Whitney U test was applied for the comparisons of RABL6 methylation levels between clinical characteristics subgroups of LC. Results : Compared to the healthy controls，the methylation of RABL6 _CpG_ 17 was inversely associated with LC in females ( per - 10% methylation : OR = 2. 47，95% CI = 1. 19－5. 13，P = 0. 016) ，but positively associated with LC in males (per - 10% methylation : OR = 0. 52，95% CI = 0. 29－0. 94，P = 0. 030) ．In addition，hypermethylation of RABL6_CpG _2 and RABL6_CpG_5 was significantly associated with LC in the subjects older than 55 years (for RABL6_CpG_ 2 : per －10% methylation : OR = 0. 77，95% CI = 0. 60－0. 99，P = 0. 038 ; for RABL6_CpG_5 : OR = 0. 58，95% CI = 0. 34－0. 97，P = 0. 038) ． Conclusion The study reveals an association between peripheral blood-based RABL6 methylation levels and early LC，providing a new clue for developing peripheral blood-based DNA methylation as a potential marker for the evaluation of LC risk.

BACKGROUNDCombined emphysema and pulmonary fibrosis, including idiopathic pulmonary fibrosis (IPF), is a distinct disorder described with upper-lobe emphysema and lower-lobe fibrosis on chest computed tomography. Smoking appears to be the predominant risk factor for this disorder. We aimed to compare clinical features, smoking history, physiological and radiological findings between IPF with and without emphysema.

METHODSA sample of 125 IPF patients over a period of 48 months were evaluated. High resolution CT scans were reviewed blinded to clinical data. The IPF patients with or without emphysema were classified accordingly.

RESULTSThe prevalence of emphysema in this IPF sample was 70/125. IPF with emphysema was significantly associated with smoking status (OR 63; 95% CI 4.4 to 915; P = 0.002) and smoking pack year (OR 1.1; 95% CI 1.05 to 1.13; P = 0.000). The patients with IPF and emphysema had a higher decrease in carbon monoxide diffusing capacity adjusted for alveolar volume ((58±19)% pred vs. (66±21)% pred; P = 0.021) and a higher prevalence of pulmonary hypertension (24/70 vs. 7/55; P = 0.006). The two groups of patients had similar forced and residual volumes. No significant differences were found in cell differentials of bronchoalveolar lavage or the scores of fibrosis on chest CT. Survival of the patients with emphysema was significantly less than that of patients with IPF alone.

CONCLUSIONSCigarette smoking induces IPF combined with emphysema. Emphysema further impairs physiological function and increases the prevalence of pulmonary hypertension that leads to poor prognosis. The inclusion of the patients with combined pulmonary fibrosis and emphysema in IPF clinical trials may lead to under evaluation of the effect of treatment in patients.

Aged ; Female ; Humans ; Idiopathic Pulmonary Fibrosis ; etiology ; physiopathology ; Male ; Middle Aged ; Pulmonary Emphysema ; complications ; physiopathology ; Smoking ; adverse effects

Animals ; CRISPR-Cas Systems ; Gene Editing ; Humans

BACKGROUND: Liver cancer is largely resistant to chemotherapy. This study aimed to identify the effective chemotherapeutics for β-catenin-activated liver cancer which is caused by gain-of-function mutation of catenin beta 1 ( CTNNB1 ), the most frequently altered proto-oncogene in hepatic neoplasms. METHODS: Constitutive β-catenin-activated mouse embryonic fibroblasts (MEFs) were established by deleting exon 3 ( β-catenin Δ(ex3)/+ ), the most common mutation site in CTNNB1 gene. A screening of 12 widely used chemotherapy drugs was conducted for the ones that selectively inhibited β-catenin Δ(ex3)/+ but not for wild-type MEFs. Untargeted metabolomics was carried out to examine the alterations of metabolites in nucleotide synthesis. The efficacy and selectivity of methotrexate (MTX) on β-catenin-activated human liver cancer cells were determined in vitro . Immuno-deficient nude mice subcutaneously inoculated with β-catenin wild-type or mutant liver cancer cells and hepatitis B virus ( HBV ); β-catenin lox(ex3)/+ mice were used, respectively, to evaluate the efficacy of MTX in the treatment of β-catenin mutant liver cancer. RESULTS: MTX was identified and validated as a preferential agent against the proliferation and tumor formation of β-catenin-activated cells. Boosted nucleotide synthesis was the major metabolic aberration in β-catenin-active cells, and this alteration was also the target of MTX. Moreover, MTX abrogated hepatocarcinogenesis of HBV ; β-catenin lox(ex3)/+ mice, which stimulated concurrent Ctnnb1- activated mutation and HBV infection in liver cancer. CONCLUSION MTX is a promising chemotherapeutic agent for β-catenin hyperactive liver cancer. Since repurposing MTX has the advantages of lower risk, shorter timelines, and less investment in drug discovery and development, a clinical trial is warranted to test its efficacy in the treatment of β-catenin mutant liver cancer.
Mice ; Animals ; Humans ; Methotrexate/therapeutic use* ; Mice, Nude ; beta Catenin/metabolism* ; Fibroblasts/metabolism* ; Liver Neoplasms/metabolism* ; Hepatitis B virus ; Nucleotides