Mechanism of total flavonoids of Ziziphora clinopodioides in improving atherosclerosis by regulating PI3K/Akt/mTOR pathway / 中国中药杂志

The present study observed the regulatory effect of total flavonoids of Ziziphora clinopodioides on autophagy and the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathways in ApoE~(-/-) mice and explored the mechanism of total flavonoids of Z. clinopodioides against atherosclerosis(AS). ApoE~(-/-) mice were fed on a high-fat diet for eight weeks to induce an AS model. The model mice were randomly divided into a model group, a positive control group, and low-, medium-and high-dose groups of total flavonoids of Z. clinopodioides, while C57BL/6J mice fed on a common diet were assigned to the blank group. The serum and aorta samples were collected after intragastric administration for 12 weeks, and the serum levels of total cholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C), and high density lipoprotein-cholesterol(HDL-C) were detected by an automatic biochemical analyzer. The serum expression levels of intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1(VCAM-1), matrix metalloproteinase-2(MMP-2), and matrix metalloprotei-nase-9(MMP-9) were detected by enzyme-linked immunosorbent assay(ELISA). Oil red O staining was used to observe the aortic plaque area in mice. Hematoxylin-eosin(HE) staining was used to observe the aortic plaque and pathological changes in mice. The expression of P62 and LC3 in the aorta was detected by the immunofluorescence method. The protein expression of LC3Ⅱ/Ⅰ, Beclin-1, P62, p-PI3K, p-Akt, and p-mTOR in the aorta of mice was detected by Western blot. The results showed that compared with the blank group, the serum levels of TC, TG, LDL-C, ICAM-1, VCAM-1, MMP-2 and MMP-9 in the model group were significantly increased(P<0.01 or P<0.05), the content of HDL-C was decreased(P<0.05), intra-aortic plaque area was enlarged(P<0.01), the expression of LC3 in the aorta was significantly down-regulated, P62 expression was up-regulated(P<0.01 or P<0.05), the expressions of LC3Ⅱ/Ⅰ and Beclin-1 in the aortic lysate were significantly down-regulated, and the expressions of p-PI3K, p-Akt, p-mTOR and P62 were significantly increased(P<0.01). The medium-and high-dose groups of total flavonoids of Z. clinopodioides could reduce the serum levels of TC, TG, LDL-C, ICAM-1, VCAM-1, MMP-2, and MMP-9 in AS model mice(P<0.01 or P<0.05), and increase the content of HDL-C(P<0.01 or P<0.05). The aortic plaque area of mice after middle and high doses of total flavonoids of Z. clinopodioides was significantly reduced(P<0.01), the content of foam cells decrease, and the narrowing of the lumen decreased. The total flavonoids of Z. clinopodioides significantly increased the expression of LC3 in the aorta and the expression of LC3Ⅱ/Ⅰ and Beclin-1 in the lysate, and decreased the expression of P62 in the aorta and the expression of p-PI3K, p-Akt, p-mTOR and P62 in the lysate(P<0.01 or P<0.05). The results showed that the total flavonoids of Z. clinopodioides could improve the content of blood lipids and inflammatory factors, and reduce the generation of foam cells and plaques in aortic tissue, and the mechanism may be related to the regulation of PI3K/Akt/mTOR signaling pathway.

The Prognostic Value of 18F-Fluorodeoxyglucose PET/CT in the Initial Assessment of Primary Tracheal Malignant Tumor: A Retrospective Study

Objective@#To investigate the potential value of 18F-fluorodeoxyglucose (FDG) PET/CT in predicting the survival of patients with primary tracheal malignant tumors. @*Materials and Methods@#An analysis of FDG PET/CT findings in 37 primary tracheal malignant tumor patients with a median follow-up period of 43.2 months (range, 10.8–143.2 months) was performed. Cox proportional hazards regression analyses were used to assess the associations between quantitative 18F-FDG PET/CT parameters, other clinic-pathological factors, and overall survival (OS). A risk prognosis model was established according to the independent prognostic factors identified on multivariate analysis. A survival curve determined by the Kaplan-Meier method was used to assess whether the prognosis prediction model could effectively stratify patients with different risks factors. @*Results@#The median survival time of the 37 patients with tracheal tumors was 38.0 months, with a 95% confidence interval of 10.8 to 65.2 months. The 3-year, 5-year and 10-year survival rate were 54.1%, 43.2%, and 16.2%, respectively. The metabolic tumor volume (MTV), total lesion glycolysis (TLG), maximum standardized uptake value, age, pathological type, extension categories, and lymph node stage were included in multivariate analyses. Multivariate analysis showed MTV (p = 0.011), TLG (p = 0.020), pathological type (p = 0.037), and extension categories (p = 0.038) were independent prognostic factors for OS. Additionally, assessment of the survival curve using the Kaplan-Meier method showed that our prognosis prediction model can effectively stratify patients with different risks factors (p 5.19, a TLG > 16.94 on PET/CT scans, squamous cell carcinoma, and non-E1 were more likely to have a reduced OS.

Chemopreventive and Chemotherapeutic Effect of Propolis and Its Constituents: A Mini-review

Propolis is a bee wax rich in various phytocomponents and traditionally used to treat various ailments. Propolis is reported to possess an array of biological properties including anti-inflammatory, antioxidant, anti-cancer, and anti-diabetic as well as cardioprotective, hepatoprotective, renoprotective, and derma protective activities. A plethora of studies confirmed that propolis is effective against various types of cancer including head and neck, lung, liver, brain (glioma), pancreas, kidney, prostate, skin (melanoma), breast, oral, esophagus, gastric, colorectal, and bladder cancers. However, many researchers have demonstrated that propolis displays potent chemoprotective/chemopreventive or anti-cancer activity against only a few types of cancers like oral, gastrointestinal, dermal (melanoma), breast, and prostate cancers. Therefore, this mini-review only summarizes the chemopreventive/chemotherapeutic activities of propolis and its updated underlying mechanisms. Taken together, propolis displays potent chemoprotective or anti-cancer effect due to the presence of various phytocomponents which contribute to pro-apoptotic, cytotoxic, anti-proliferative (cell cycle arrest), anti-metastatic, anti-invasive, anti-angiogenic and anti-genotoxic or anti-mutagenic properties along with antioxidant, immunomodulatory, and anti-inflammatory functions. Hence, propolis could be used as an adjuvant for treating various cancers along with standard chemotherapeutic drugs. However, many large-scale clinical studies are needed to justify such applications.

Clinical Efficacy of Xiaoer Anorexia Granule Combined with Saccharomyces Brassicus in Treatment of Pediatric Anorexia and Its Effect on Serum Cytokines / 中国实验方剂学杂志

Objective: To investigate the clinical efficacy of Xiaoer anorexia granule combined with Saccharomyces brassicus in the treatment of infantile anorexia and its effect on serum cytokines. Method: Totally 240 cases of infantile anorexia treated in Jinhua People's Hospital from October 2015 to October 2017 were randomly divided into three groups. Group A (combination group) orally took Xiaoer anorexia granule, 1 pack/time for 1-3 years old, 2 packs/time for 4-6 years old, 3 times a day, and combined with saccharomyces brasiliensis, 0.5 g/time, 2 times a day, 15-30 min before meal. Group B took saccharomyces brasiliensis by the same method as the combination group. Group C took Xiaoer anorexia granules by the same method as above,with 80 cases in each group. 8 weeks is a course of treatment. Serum inflammatory cytokines leptin, SP and Ghrelin were also detected. The clinical efficacy, body weight, height improvement and serum cytokine levels were observed in 3 groups. Result: The total improvement rate of group A was 91.25%, which was significantly higher than 71.25%of group B and 73.75%of group C, with statistically significant differences (PPPPPPPConclusion: Treatment of infantile anorexia with Xiaoer anorexia granules combined with Saccharomyces cerevisiae can improve the total clinical effective rate, adjust serum cytokine levels and increase height and body mass.

Inhibitory effects of petasin on human colon carcinoma cells mediated by inactivation of Akt/mTOR pathway / 中华医学杂志(英文版)

BACKGROUND@#Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far. Petasin, a natural product found in plants of the genus Petasites, has been reported to possess anticancer activity. The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo. The molecular mechanism of petasin was also further explored.@*METHODS@#Caco-2, LoVo, SW-620, and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation. Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis was analyzed by flow cytometry. Hoechst 33258 staining was used to visualize morphological changes. Cell migration was assessed using a wound-healing migration assay, and cell invasion was investigated using Transwell chambers. Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway. Finally, in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice. Twelve rats were randomly divided into control group and 10 mg/kg petasin group. The tumor volume was calculated every 7 days for 28 days. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin. Differences between two groups were assessed by analysis of independent-sample t tests.@*RESULTS@#Petasin significantly inhibited the proliferation of human colon carcinoma cell lines, induced apoptosis, and suppressed migration and invasion in SW-620 cells. Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs. 0.74 ± 0.06, P = 0.042), mTOR (0.71 ± 0.12 vs. 0.32 ± 0.11, P = 0.013), and P70S6K (1.23 ± 0.21 vs. 0.85 ± 0.14, P = 0.008), elevated the expression of caspase-3 (0.41 ± 0.09 vs. 0.74 ± 0.12, P = 0.018) and caspase-9 (1.10 ± 0.27 vs. 1.98 ± 0.22, P = 0.009), decreased the Bcl-2 protein (2.75 ± 0.47 vs. 1.51 ± 0.36, P = 0.008), downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs. 0.82 ± 0.11, P = 0.021) and MMP-9 (1.56 ± 0.32 vs. 0.94 ± 0.15, P = 0.039) in SW-620 cell. In vivo, 10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs. 577.67 ± 75.12 mm at day 28, P = 0.001) and induced apoptosis (3.6 ± 0.7% vs. 36.0 ± 4.9%, P = 0.001) in tumor tissues.@*CONCLUSIONS@#Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway. Our findings suggest petasin as a potential candidate for colon cancer therapy.

The significance of eosinophils in the correlation of upper and lower airway inflammation in patients with chronic rhinitis / 中华耳鼻咽喉头颈外科杂志

Objective@#To explore the predictor of lower airway inflammation among the index of nasal inflammation by investigating the expression and association of eosinophils (EOS) in the upper-lower airways and blood of patients with chronic rhinitis.@*Methods@#A total of 162 patients with allergic rhinitis (AR), 117 patients with non-allergic rhinitis (NAR) and 104 controls were enrolled from June 2010 to December 2013 from General Hospital of Eastern Theater Command, People′s Liberation Army. All subjects were required detailed medical history collection and nasal resistance measurement. Skin prick test (SPT), blood total immunoglobin E (tIgE) and blood EOS, nasal lavage and induced sputum EOS, nasal provocation and bronchial provocation test (NPT, BPT), nasal and forced exhaled nitric oxide (NNO, FeNO) were performed in all patients. One-way analysis of variance was used for comparison between groups. LSD t test or Mann-Whitney U test was used for comparison between the two groups. Pearson or Spearman related parameter test was used for correlation analysis.@*Results@#The nasal lavage EOS, NNO, induced sputum EOS, FeNO, blood EOS and tIgE were higher in the AR group than that in the NAR group (3.70[1.20, 14.23]/200 HP vs 1.40[0.20, 3.40]/200 HP, 673.50[466.80, 936.00] ppb vs 455.80[248.10, 705.60] ppb, 2.97[0.00, 10.63]% vs 1.00[0.23, 2.00]%, (49.28±26.37)ppb vs (34.07±19.11)ppb, 4.00[2.00, 7.00]% vs 2.00[1.00, 5.00]%, 208.01[61.70, 387.50] IU/ml vs 43.30[19.00, 122.00] IU/ml, F or χ2 value was 11.442, 19.440, 70.727, 69.449, 47.453, 46.525, respectively, all P<0.05). But there was no significant difference in nasal resistance, NPT and BPT between the two groups. Nasal lavage EOS in AR group and NAR group was correlated with induced sputum EOS, FeNO, tIgE and blood EOS (r value of AR group was 0.448, 0.202, 0.159, 0.321, r value of NAR group was 0.442, 0.268, 0.268, 0.334, respectively, all P<0.05), but not with BPT. After adjustment for gender, age, height and weight, nasal EOS was positively correlated with sputum EOS. Multiple linear regression analysis showed that nasal EOS, blood EOS and SPT were factors affecting sputum EOS levels. The optimal threshold for nasal EOS to determine induced sputum EOS was 3.30/200 HP by (receiver operating characteristic,ROC) analysis.@*Conclusion@#The nasal EOS is correlated with multiple lower airway and systemic inflammatory markers, and is a risk factor for the induced sputum EOS, which can be used as an inflammation biomarker to predict the lower air inflammation.

Inhibitory effects of petasin on human colon carcinoma cells mediated by inactivation of Akt/mTOR pathway / 中华医学杂志(英文版)

Background:@#Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far. Petasin, a natural product found in plants of the genus Petasites, has been reported to possess anticancer activity. The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo. The molecular mechanism of petasin was also further explored.@*Methods:@#Caco-2, LoVo, SW-620, and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation. Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis was analyzed by flow cytometry. Hoechst 33258 staining was used to visualize morphological changes. Cell migration was assessed using a wound-healing migration assay, and cell invasion was investigated using Transwell chambers. Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway. Finally, in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice. Twelve rats were randomly divided into control group and 10 mg/kg petasin group. The tumor volume was calculated every 7 days for 28 days. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin. Differences between two groups were assessed by analysis of independent-sample t tests.@*Results:@#Petasin significantly inhibited the proliferation of human colon carcinoma cell lines, induced apoptosis, and suppressed migration and invasion in SW-620 cells. Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs. 0.74 ± 0.06, P = 0.042), mTOR (0.71 ± 0.12 vs. 0.32 ± 0.11, P = 0.013), and P70S6K (1.23 ± 0.21 vs. 0.85 ± 0.14, P = 0.008), elevated the expression of caspase-3 (0.41 ± 0.09 vs. 0.74 ± 0.12, P = 0.018) and caspase-9 (1.10 ± 0.27 vs. 1.98 ± 0.22, P = 0.009), decreased the Bcl-2 protein (2.75 ± 0.47 vs. 1.51 ± 0.36, P = 0.008), downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs. 0.82 ± 0.11, P = 0.021) and MMP-9 (1.56 ± 0.32 vs. 0.94 ± 0.15, P = 0.039) in SW-620 cell. In vivo, 10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs. 577.67 ± 75.12 mm3 at day 28, P = 0.001) and induced apoptosis (3.6 ± 0.7% vs. 36.0 ± 4.9%, P = 0.001) in tumor tissues.@*Conclusions:@#Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway. Our findings suggest petasin as a potential candidate for colon cancer therapy.

Comparison of efficacy and safety between two different methods of nephroureterectomy in two centers / 北京大学学报(医学版)

OBJECTIVE@#To compare the efficacy and safety of complete transperitoneal laparoscopic nephroureterectomy (CTNU) and traditional retroperitoneoscopic nehroureterectomy (TRNU) for the management of upper urinary tract urothelial carcinoma(UTUC).@*METHODS@#We retrospectively collected the clinical data of UTUC patients who underwent CTNU or TRNU surgery from January 2011 to December 2018 in Peking University First Hospital and Fujian Provincial Hospital, and compared the clinical characteristics, perioperative parameters, and follow-up results between the CTNU and TRNU surgeries.@*RESULTS@#Finally, a total of 266 cases were included, with 94 cases in the CTNU group and 172 cases in the TRNU group. The proportion of left side lesions was bigger in TRNU group when compared with CTNU group (P<0.05). No significant differences were observed in clinical characteristics, such as age, gender, body mass index (BMI), American society of anesthesiologists score (ASA score) and tumor laterality. All surgery procedures were completed. The vascular resparing was performed by reason that left arteria renalis was injured accidently during surgical operation in one case of TRNU group. No serious complications were observed in both CTNU and TRNU groups. In CTNU group, operating time was (202.9±76.7) min, estimated blood loss was (68.4±73.3) mL, drainage duration was (3.9±1.5) d, drainage volume was (181.7±251.5) mL, and postoperative hospital stay was (7.8±4.1) d. In TRNU group, operating time was (203.5±68.7) min, estimated blood loss was (130.2±252.1) mL, drainage duration was (4.3 ±1.6) d, drainage volume was (179.1±167.5) mL, and postoperative hospital stay was (8.2±3.7) d. The estimated blood loss in CTNU group was significantly less than that in TRNU group (P=0.005).The median follow-up time was 39 months (range: 1-88 months). The 5-year overall survival rate (OS), cancer specific survival rate (CSS), intra-vesical recurrence free survival rate (IvRFS), disease free survival rate (DFS) of CTNU group was 75.6%, 86.9%, 73.8%, 57.5%, respectively. The OS, CSS, IvRFS and DFS of TRNU group was 66.3%, 83.5%, 75.9%, 58.6%, respectively.No significant differences were observed in the OS, CSS, IvRFS and DFS between the CTNU and TRNU groups.@*CONCLUSION@#CTNU technique is a safe and effective surgical option, and further prospective randomized controlled trial is needed for further evaluation.

Relationship between iASPP expression with clinicopathological features and prognosis in patients with nasopharyngeal carcinoma / 重庆医学

Objective To investigate the expression change of inhibitory member of the ASPP family(iASPP) and its relationship with clinicopathological features and prognosis in the patients with nasopharyngeal carcinoma(NPC).Methods One hundred and thirty-three cases of nasopharyngeal carcinoma confirmed by pathology in the hospital were selected,at the same time 30 cases of nasopharyngeal chronic mucositis biopsy paraffin blocks previously collected in the pathology department served as the control group.The immunohistochemical staining was used to compare the expression of iASPP protein in the two groups and the relationship between the expression of iASPP with the clinicopathologic characteristics and prognosis in the patients with NPC was analyzed.Results The positive expression of iASPP protein in the nasopharyngeal carcinoma tissue was 72.18％,which was higher than 20.00％ in the control group(P＜0.05);the positive expression of iASPP protein in the nasopharyngeal carcinoma tissue had a certain relationship with the primary tumor range and lymph node metastasis(P＜0.05);92 cases of nasopharyngeal carcinoma were followed up,including 63 cases of iASPP protein positive expression and 29 cases of iASPP protein negative expression,the 5-year survival rate in the patients with iASPP protein positive expression was 71.43％,which in the patients with iASPP protein negative expression was 86.21％,the difference was not statistically significant(P＞0.05).Conclusion The iASPP expression in the nasopharyngeal carcinoma tissue is up-regulated,which is related to the lymph node metastasis and the primary tumor range,but its influence on the patient's prognosis is not significant.

The role of NF-κB signaling pathway in laryngeal leukoplakia recurrent with laryngeal reflux / 中华耳鼻咽喉头颈外科杂志

Objective@#To study the mechanism of vocal mucosal barrier damage mediated by NF-κB and NF-κB-regulated signaling pathway via probing the expression of inflammatory factors and essential proteins for node of NF-κB signaling pathway.@*Methods@#The patients suffering from vocal leukoplakia accompanied with larygopharyngeal reflux(LPR) were treated with oral administration of proton pump inhibitor(PPI). Mucosal specimens of vocal cord were collected from all patients before PPI treatment. And the mucosal specimens of vocal cord were collected from the patients with suspected recurrence at 8 weeks after PPI treatment. HE staining was used to observe the histopathological changes of the mucosa. ELISA was utilized to detect the levels of inflammatory factors including tumor necrosis factor(TNF)-α, interleukin(IL)-1 and IL-6. Western blot was used to detect the expression of p-p65, p-IKK and p-IκB. Immunofluorescence method was adopted to detect the entrance of p65 to cell nucleus.Data was analyzed by SPSS 23.0 software.@*Results@#In PPI untreated group, the expressions of IL-1β, TNF-α and IL-6 in the specimens of 8 weeks after operation were not different significantly from those obtained during operation.But in the PPI-treated group, the expressions were down-regulated.The expression of p-p65 in the middle and high grade heterogenous hyperplasia group was higher than that of low level heterogenous hyperplasia group.The difference of p65 and p-p65 expression between 8 weeks after surgery and surgery in PPI-untreated group was statistically insignificant (P>0.05). The difference of p65 expression between PPI-treated group and PPI pre-treatment group was statistically insignificant (P>0.05). The expression of p-p65 in the PPI-treated group was lower than that of the PPI pre-treatment group (P<0.05). The expressions of IL-1β, TNF-α and IL-6 were positively related with that of NF-κB-p65. Immun of luorescence method revealed the entrance of p65 to cell nucleus in PPI pre-treatment group, which meant that NF-κB was activated. In the PPI-treated group, few activated p65 could be observed in the cell nucleu.@*Conclusion@#The possible mechanism of vocal mucosal barrier damage in vocal leukoplakia accompanied with LPR maybe the vocal mucosal inflammation mediated by NF-κB and NF-κB-regulated signaling pathway activated with refluxed materials.

In vitro antibacterial activity analysis of antibacterial peptide lycosin-Ⅰ against clinical isolates of Pseudomonas aeruginosa / 临床检验杂志

Objective To investigate the in vitro antibacterial activity of lycosin-Ⅰ against clinical isolates of Pseudomonas aeruginosa.Methods The clinical isolates of Pseudomonas aeruginosa,including 10 strains of multidrug resistant Pseudomonas aeruginosa and 10 strains of non-multidrug resistant Pseudomonas aeruginosa,were randomly collected from the Second Xiangya Hospital of Central South University.The in vitro minimum inhibitory concentrations (MIC) of lycosin-Ⅰ against Pseudomonas aeruginosa were detected by the broth microdilution method.The representative isolates of multidrug resistant (Isolate 8,MIC =8 μg/mL) and non-multidrug resistant (Isolate 12,MIC =8 μg/mL) Pseudomonas aeruginosa were selected,and the bactericidal kinetics of 4 × MIC concentration of lycosin-Ⅰ against them were determined.After the representative isolates were cultured with suitable medium for 24 hours,the absorbency of 600 nm was detected and the growth curve was drawn.Five mmol/L of Ca2+ or Mg2+ were added into the medium to investigate the salt tolerance of lycosin-Ⅰ.Results Lycosin-Ⅰ in vitro showed good antibacterial activity against both multidrug resistant and non-multidrug resistant Pseudomonas aeruginosa.The MICs (median [P25,P75]) of lycosin-Ⅰ against multidrug-resistant and non-multidrug-resistant Pseudomonas aeruginosa were 12 (8,32) μg/mL,and there was no statistical difference between them (U =42,P ＞0.05).About 50％ of multidrug resistant and non-multidrug resistant Pseudomonas aeruginosa could be killed by 4 × MIC of lycosin-Ⅰ during 60 minutes.When the concentration of lycosin-Ⅰ below MIC,such as 0,2 or 4 μg/mL,was co-cultured with multidrug resistant and non-multidrug resistant Pseudomonas aeruginosa for 24 hours,the growth of bacteria was fast during 6 and 16 hours,and then tended to be slow after 18 hours.When the concentration of lycosin-Ⅰ increased to 8 μg/mL,the bacteria were hard to grow.The in vitro antibacterial activity of lycosin-Ⅰ could be reduced by the addition of 5 mmol/L of Ca2+ or Mg2+,and the MICs of lycosin-Ⅰ against multidrug resistant and non-multidrug resistant Pseudomonas aeruginosa increased from 8 μg/mL to 64 μg/mL for 5 mmol/L of Ca2+,and from 8 μg/mL to 32 μg/mL for 5 mmol/L of Mg2+,respectively.But high concentrations of lycosin-Ⅰ (＞ 64 μg/mL or ＞ 32 μg/mL) could still maintain its antibacterial activity against multidrug resistant and non-multidrug resistant Pseudomonas aeruginosa.Conclusion Lyosin-Ⅰ can effectively inhibit the growth of Pseudomonas aeruginosa in vitro,and has certain salt tolerance,which may be developed into a new type of antibacterial drug.

Regulatory mechanism of UVB-induced autophagy and apoptosis in A375 cells / 中国职业医学

OBJECTIVE: To investigate the effect of ultraviolet B( UVB) on autophagy and apoptosis in human epidermal melanoma A375 cells. METHODS: i) A375 cells at logarithmic growth phase were exposed to UVB at doses of 10. 0 and15. 0 m J/cm~2. Then cells were collected at time point of 3,6,9 and 12 hours after irradiation. The effect of UVB on cell autophagy was observed by monodansylcadaverine staining and the effect of UVB on cell apoptosis was observed by acridine orange/ethidium bromide staining. ii) A375 cells of 10. 0 m J/cm~2 group and 15. 0 m J/cm~2 group were exposed to corresponding dose of UVB irradiation. Then cells were collected at time point of 18,24,36 and 48 hours after irradiation,and cell survival rate was examined using CCK-8 assay. iii) A375 cells were irradiated with UVB at doses of 10. 0 and15. 0 m J/cm~2 and then cells were collected at time point of 3,6,9 and 12 hours after irradiation. After that,A375 cells were irradiated at doses of 2. 5,5. 0,7. 5,10. 0 and 15. 0 m J/cm~2 of UVB,then cells were collected at time point of 9 hours after irradiation. The expressions of B-lymphoblastoma-2( Bcl-2),Bcl-2 related X protein( Bax),Bcl-2 interacting protein( Beclin-1) and microtubule-associated protein 1 light chain 3( LC3) Ⅱ were detected by Western blotting. A375 cells with no UVB irradiation were set as the control( pseudo-irradiation) in each experiment. RESULTS: i) Both autophagy and apoptosis of A375 cells induced by UVB irradiation at doses of 10. 0 and 15. 0 m J/cm~2 increased with time after irradiation. The effect on autophagy decreased at 12 hours time point with 15. 0 m J/cm~2 UVB irradiation. ii) The cell viability increased with time after irradiation in the 10. 0 and 15. 0 m J/cm~2 groups( P < 0. 05). From 18-48 hours after irradiation,the cell viability of the 10. 0 and 15. 0 m J/cm~2 groups was lower than that of the control group( P < 0. 05).From 24-48 hours after irradiation,the cell viability of the 15. 0 m J/cm~2 group was lower than that of the 10. 0 m J/cm~2 group( P < 0. 05). iii) The relative expression of Beclin-1 and LC3 Ⅱ protein at the 10. 0 m J/cm~2 group increased with time after 0-12 hours irradiation( P < 0. 05). The above changes of the 15. 0 m J/cm~2 group were observed within 0 to 9 hours,and the above two autophagy-related proteins were significantly decreased at the 12 hours time point( P < 0. 05).The relative expression of Bcl-2 protein at the 10. 0 and 15. 0 m J/cm~2 groups decreased with increasing time from 3 to 12 hours after irradiation( P < 0. 05). The relative expression of Bax protein increased with time from 0 to 12 hours after irradiation( P < 0. 05). The relative expression of Beclin-1 and LC3 protein in cells at 0. 0-10. 0 m J/cm~2,and the relative expression of Bax protein in cells at 0. 0-15. 0 m J/cm~2 increased with increase of irradiation dose( P < 0. 05). The relative expression of Bcl-2 protein decreased with increase of irradiation dose at 5. 0-15. 0 m J/cm~2( P < 0. 05). CONCLUSION: Autophagy and apoptosis of A375 cells can be induced by UVB irradiation at doses of 10. 0 and 15. 0 m J/cm~2. Autophagy induced by UVB irradiation at 10. 0 m J/cm~2 partially resisted the induction of apoptosis by UVB and enhanced cell viability. 15. 0 m J/cm~2 UVB-induced autophagy was insufficient to exert the above-mentioned effects,and the induction of apoptosis was the dominant effect.

Clinical analysis and treatment of nasal complications after transsphenoidal pituitary adenomasectomy / 中国耳鼻咽喉头颈外科

OBJECTIVE To exlpore the prevention and therapy methods of nasal complications after transsphenoidal pituitary adenomasectomy.METHODS 129 cases of pituitary adenomas underwent transsphenoidal surgery.The postoperative nasal complications were observed and disposed.RESULTS A total of 26 cases(20.1％) of nasal complications appeared in all 129 cases of pituitary adenomas underwent transsphenoidal surgery,including nasal hemorrhage 6 cases(4.8％),eerebrospinal fluid rhinorrhea 9 cases(6.9％),sphenoid sinusitis 3 cases(2.3％),atrophy rhinitis 2 cases(1.6％),olfactory dysfunction 2 cases(1.6％),nasal septum perforation 1 case(0.8％),and nasal adhesion 3 cases(2.3％).They were all cured after nasal endoscopic regular cavity clean,nasal hormone,nasal irrigation.CONCLUSION The nasal complications after transsphenoidal pituitary adenomas ectomy surgery should be timely disposed,which can effectively improve the patients' clinical symptoms.

Development of Tianma HPLC fingerprint and discriminant analysis / 中国中药杂志

Tianma(the tuber of Gastrodia eleta) is a widely used and pricy Chinese herb. Its counterfeits are often found in herbal markets, which are the plant materials with similar macroscopic characteristics of Tianma. Moreover, the prices of Winter Tianma(cultivated Tianma) and Spring Tianma(mostly wild Tianma) have significant difference. However, it is difficult to identify the true or false, good or bad quality of Tianma samples. Thus, a total of 48 Tianma samples with different characteristics(including Winter Tianma, Spring Tianma, slice, powder, etc.) and 9 plant species 10 samples of Tianma counterfeits were collected and analyzed by HPLC-DAD-MS techniques. After optimizing the procedure of sample preparation, chromatographic and mass-spectral conditions, the HPLC chromatograms of all those samples were collected and compared. The similarities and Fisher discriminant analysis were further conducted between the HPLC chromatograms of Tianma and counterfeit, Winter Tianma and Spring Tianma. The results showed the HPLC chromatograms of 48 Tianma samples were similar at the correlation coefficient more than 0.848(n=48). Their mean chromatogram was simulated and used as Tianma HPLC fingerprint. There were 11 common peaks on the HPLC chromatograms of Tianma, in which 6 main peaks were chosen as characteristic peaks and identified as gastrodin, p-hydroxybenzyl alcohol, parishin A, parishin B, parishin C, parishin E, respectively by comparison of the retention time, UV and MS data with those of standard chemical compounds. All the six chemical compounds are bioactive in Tianma. However, the HPLC chromatograms of the 10 counterfeit samples were significantly different from Tianma fingerprint. The correlation coefficients between HPLC fingerprints of Tianma with the HPLC chromatograms of counterfeits were less than 0.042 and the characteristic peaks were not observed on the HPLC chromatograms of these counterfeit samples. It indicated the true or false Tianma can be identified by either the similarity or characteristic peaks on HPLC fingerprint. Comparing the Winter Tianma with Spring Tianma showed that the HPLC chromatograms of 15 winter Tianma samples and 11 spring Tianma samples were similar at the mean correlation coefficient of 0.908. But the intensity of the characteristic peaks were different between the two groups of Tianma samples, i.e. the intensity of gastrodin, paishin A and C in winter Tianma was lower than those in spring Tianma. The Winter Tianma and Spring Tianma could be discriminated by either the Fisher unstandardized discrimination function or Linear discriminant function, based on the peak areas of 11 common peaks on HPLC chromatograms as variate.

Changes in small airway function in rhinitis without asthma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Observe the changes of small airway function in patients with rhinitis but without asthma and/or lower airway symptoms.@*METHOD@#Between June 2008 and December 2012, we recruited 903 subjects, including 377 with allergic rhinitis (AR), 262 with non-allergic rhinitis (NAR) and 264 healthy subjects. All subjects underwent meticulous history taking, nasal examination, allergen skin prick test, blood routine test, serum total immunoglobin E assay, pulmonary ventilation function test and bronchial challenge test.@*RESULT@#The indices of FEV1/FVC%, MEF25pred% and MMEFpred% were lower in AR group than in the control group (P 0.05). The positive rate of airway hyperresponsiveness(AHR) in AR group and in NAR group was 12.2%, 6.1% respectively. Indices of small airway function were all lower in the AHR group than NAHR group in rhinitis.@*CONCLUSION@#Compared with healthy controls, small airway function in patients with rhinitis has apparent changes, part of rhinitis patients has AHR, and is associated with small airway function changes.

Effects of allergic and nonallergic rhinitis on bronchial inflammation / 医学研究生学报

Objective To describe the effects of allergic and nonallergic rhinitis on patients with bronchial inflammation by comparing the bronchial inflammation characteristics . Methods All patients enrolled in the study were categorized into two groups according to rhinitis diagnostic criteria , 377 cases in the allergic rhinitis ( AR) group, 262 cases in non-allergic rhinitis ( NAR) group and another 264 patients without rhinitis as control group .Induced sputum differential cytology tests , exhaled nitric oxide concentration measurements and methacholine bronchial provocation tests were performed .The characteristics and differences in lower airway inflam-mation among the three groups were compared . Results The exhaled nitric oxide level and the positive rate of the exhaled nitric ox-ide were different significantly in AR group and NAR group than the control group (P0.05). Conclusion Bronchial inflammation may be aggravated by AR and NAR , so all patients with AR and NAR need treatment , and the degree of inflammation was higher in AR patients than the NAR patients .

Correlation between contents of gastrodin and polysaccharides with grade of Gastrodia Tuber / 中草药

Objective: To study the correlation between the contents of gastrodin and polysaccharides with the variety of commercial specifications, grades, and cultivated areas of Gastrodia Tuber (Tianma), the tuber of Gastrodia elata, and then to choose the suitable chemical marker to assess the quality of Tianma raw materials. Methods: Gastrodin and polysaccharides were quantified in 35 Tianma samples including Winter Tianma which was harvested in winter, Spring Tianma which was harvested in spring with their different grades and cultivation areas by HPLC and spectrophotometry, respectively. Combined with the morphological characteristics, the correlation between the various Tianma samples with the amounts of gastrodin and polysaccharides was studied. Results: The content of polysaccharides in Winter Tianma (26.05%, n = 24) was higher than that in Spring Tianma (20.21%, n = 10). In contrast, the amount of gastrodin in Winter Tianma (0.49%, n = 24) was lower than that in Spring Tianma (0.68%, n = 10). Among Tianma samples with different grades, the content of polysaccharides in the first grade (32.65%, n = 6) and second grade (32.89%, n = 6) was higher than that in the third grade (21.71%, n = 8) and fourth grade samples (14.57%, n = 4). But, the amounts of gastrodin in the Tianma samples of the four grades were similar, which were 0.46% (n = 6), 0.65% (n = 6), 0.43% (n = 8), and 0.44% (n = 4) in the first, second, third, and fourth grade Tianma, respectively. Moreover, the contents of gastrodin and polysaccharides in Tianma samples collected from the famous cultivation regions such as Sichuan, Shaanxi, and Yunnan provinces of China were higher than those from others areas. Conclusion: The developed HPLC and spectrophotometry methods are accurate and reliable to assay the amounts of gastrodin and polysaccharides in Tianma. The amount of polysaccharides is related to the quality of Tianma samples authenticated by the commercial specifications, grades, and traditional experience, and therefore is a better marker to assess the quality of Tianma.

Distribution of HCV genotypes in Chinese Han population with chronic hepatitis C / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the distribution of HCV genotypes in Chinese Han population with chronic hepatitis C (CHC).</p><p><b>METHODS</b>This randomized multicenter study included 1 014 CHC patients from 28 hospitals in different regions of China. SPSS 20.0 was applied to analyze the relationship among region, HCV genotype, gender and the replication level of HCV-RNA.</p><p><b>RESULTS</b>HCV 1 genotype (56.80%) was the most common genotype. The majority of CHC patients were of genotype 1, 2, 3, 6 in the order of frequency, except those in southwestern, southern and central China. HCV 1, 2, 3, 6 genotypes were most common among male patients in southern China; among female patients in northern China; among male patients in northern and northwestern China and among male patients in northwestern China, respectively (all P <0.05). There was no statistical significance between different genders in other regions. The high viral load was more common than the low viral load among HCV 1, 2, 3, 6 genotype-infected patients.</p><p><b>CONCLUSION</b>There are different distributions of HCV genotypes among the different regions. In addition, HCV genotypes are correlated with gender and HCV-RNA load.</p>

Changes in small airway function in patients with non-allergic rhinitis / 医学研究生学报

Objective Small airway hypofunction is an early manifestation of asthmatic airway injury and is found in patients with non-asthma allergic rhinitis.However, no report has been seen on the changes of small airway function in patients with non-aller-gic rhinitis ( NAR) .This study was to investigate the possibility of small airway lesion in NAR patients and its relationship with airway responsiveness by observing the changes of small airway function in NAR patients without asthma and/or lower airway symptoms. Methods We recruited 324 subjects for this study, including 262 NAR patients and 62 healthy controls, and assigned them to an air-way hyperresponsiveness (AHR) and a non-airway hyperresponsiveness (nAHR) group.All the subjects underwent medical history collection, nasal examination, allergen skin prick test, blood routine test, serum total immunoglobin E assay, pulmonary function test, and bronchial challenge test. Results Compared with the healthy con-trols, the NAR patients showed remarkably lower predicted percenta-ges of maximal mid-expiratory flow ([85.6 ±17.1] vs [81.3 ± 19.9]%), mid-expiratory flow (MEF) with 75% of forced vital ca-pacity (FVC) expired ([96.1 ±16.1] vs [88.8 ±23.1]%), MEF with 50%of FVC expired ([88.4 ±17.8] vs [84.8 ±20.8]%), and MEF with 25%of FVC expired ([92.7 ±25.9] vs [82.9 ± 28.7]%) (P0.05).The positive rate of AHR was 6.1% (16/246) in the NAR group.All the indices of small airway function were significantly lower in the AHR than in the nAHR group (P <0.01). Conclusion NAR patients are apt to undergo obvious changes in small airway function, some with AHR, which is associated with lower airway function changes.

The effect of nasal irrigation in the treatment of allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To compare the symptoms and lower airway inflammatory factors of patients with allergic rhinitis (AR), and to observe the effect of nasal irrigation in the treatment of allergic rhinitis.@*METHOD@#Sixty-one cases diagnosed as AR after skin prick test (SPT)were selected and randomly divided into three groups: 17 patients in group A used nasal steroid spray; 21 cases in group B used nasal irrigation; 23 patients in group C combined ir rigation with nasal steroid. Before and after 3 months treatment, nasal visual analogue scale (VAS) score, rhino conjunctivtis quality of life questionnaire (RQLQ) score, fractional exhaled nitric oxide (FENO) values were observed and compared in each group.@*RESULT@#Before treatment, there is no statistically difference between three groups (P > 0.05). After 3 months of treatment, VAS, RQLQ, FENO of all patients was significantly decreased (P 0.05), FENO value has no statistically significant difference between group A and group B (P > 0.05), but were less than that in group C (P < 0.05).@*CONCLUSION@#Nasal irrigation can ameliorate nasal symptoms, improve quality of life, decrease lower airway inflammation of allergic rhinitis patients. Nasal irrigation is an effective treatment of allergic rhinitis. Nasal irrigation combined with nasal steroid can achieve more significant efficacy.