Beneficial effects of naringenin and morin on interleukin-5 and reactive oxygen species production in BALB/c mice with ovalbumin-induced asthma

We investigated the effects of naringenin and morin on IL-5 and ROS production in PMA+ionomycin-treated EL-4 cells with the corroboration of their antioxidant and anti-inflammatory properties using an asthma-induced mouse model. The EL-4 cell line was used to study the outcomes of naringenin or morin, followed by cell viability studies. Western blot analysis and ELISA test were used to determine Th2 mediated cytokines. In vivo studies were carried out on BALB/c mice to induce allergic asthma using ovalbumin administered intraperitoneally. Intracellular ROS was determined using 2’,7’-dichlorodihydrofluorescein diacetate, followed by serum enzymatic (AST and ALT) estimations and inflammatory cell count in the bronchoalveolar lavage fluid (BALF) and lung tissues. Histopathological studies were conducted to examine lung tissue-stained architecture. Our findings suggested that naringenin and morin significantly suppressed IL-5 and ROS production via various pathways. Interestingly, by reducing NFAT activity, naringenin and morin stimulated HO-1 expression, thereby suppressing IL-5 secretion due to regulating the transcription factor Nrf2 via P13/Akt or ERK/JNK signalling pathways in EL-4 cells, demonstrating the involvement of HO-1 expression in inhibiting asthmatic inflammation. The increased inflammatory cells in the BALF were substantially decreased by both naringenin and morin, followed by inhibition in the elevated Th-2 cytokines levels. The TNF-α protein levels in an allergic asthma mouse model were significantly reduced by suppressing Akt phosphorylation and eosinophil formation. Recent findings confirmed that naringenin and morin possess the potential to control asthma-related immune responses through antioxidant and anti-inflammatory properties, indicating potential therapeutic agents or functional foods.

Discussion on the sharing mechanism of cerebration of constructing the sharing platform for respiratory disease biobank / 中华医学科研管理杂志

Objective:Standardized sample resources and high-quality clinical big data are important resources for medical research, only through resource sharing can maximize its utilization.Which can be utilized to the max only through resource sharing.Methods:This paper attempts to explore the sharing mechanism of the resource sharing platform and proposes some aspects such as the platform construction background, management regulations, legal ethical system, data sharing principles, benefit distribution, etc.This article attempts to explore the sharing mechanism based on the resource sharing platform of the respiratory disease biobank, proposes the contents that should be included in the sharing mode.Detailed information including the platform construction background, management procedures, legal and ethical system, data sharing principles and benefit distribution should take into consideration in the operating mechanism of the platform.Results:Establishing a resource sharing platform matches the development of clinical research in China.The tailored sharing model which is suitable for the field of respiratory diseases will also guide the rapid development of clinical research.Conclusions:The construction of a respiratory disease biobank sharing platform is conducive to promoting the opening and sharing of biological samples and information resources in the context of big data.

Construction of recombinant PRRSV ORF5 gene by DNA shuffling technique / 生物工程学报

Recombinant PRRSV △2ORF5 gene was constructed using DNA shuffling from four genetically different strains of PRRSV to study its heterologous cross-neutralizing ability. The △2ORF5 mutant gene was cloned into the vector pET-32a and transferred into E. coli BL21. SDS-PAGE confirmed that the molecular weight of the recombinant △2ORF5 was about 42 kDa, consistent with the predicted result. Then the purified recombinant protein was injected into BALB/c mouse to obtain polyclonal antibody. Western blotting analysis with mouse-anti-△2ORF5 polyclonal serum indicated that the parental virus recombinant GP5 protein reacted with the specific antibodies. Four parental viruses could be inhibited by the anti-△2ORF5 polyclonal antibody and the inhibition rates were higher than 53%. This work has laid a foundation for further development vaccine for PRRSV.

Optimizing the Ultrasonic Extraction of Gardenia Oils from Gardenia Jasminoides Ellis by Response Surface Methodology / 世界科学技术-中医药现代化

This study aimed to optimize the ultrasonic extraction of gardenia oils from Gardenia jasminoides Ellis.by response surface analysis methodology (RSM).The duration of extraction,ratio of liquid to materials and extracting times were selected as impact factors based on single-factor experiment.RSM was adopted to learn the effects of the three factors on the extraction rate of Gardenia oil from Gardeniajasminoides Ellis.As a result,it was found that the duration of extraction was 28.7 min,while the ratio of liquid to materials was 11.62:1,and the number of extracting times were 3.36.Under this extracting condition,the average extraction rate of the oil was 13.5％ according to three validation experiments,fitting the predicted value well.In conclusion,it was demonstrated that the ultrasonic extraction method effectively improved the extraction rate of gardenia oil,providing a certain basis for the development and utilization of gardenia oil.

Survival properties of ETEC surface-displayed K88ac-LT(B) on Lactobacillus casei / 生物工程学报

K88ac-LT(B) gene derived from pQE30-K88ac-LT(B) was cloned into the expression vector pLA and then the recombinant vector was transformed into the competent cells Lactobacillus casei 525. The recombinant bacteria were grown at 37 degrees C, in MRS broth. Western blotting analysis with rabbit-anti-K88ac-LT(B) polyclonal serum indicated that the recombinant protein reacted with the specific antibodies. The results showed that the molecular weight of the recombinant protein was about 71.2 kD. The K88ac-LT(B) fusion protein on the cell surface was confirmed by immunofluorescence mciroscopy and flow cytometric analysis. In addition, the survival of recombinant Lactobacillus casei 525 was studied in imitative gastrointestinal environments such as artificial gastro fluid (pH 1.5-5.5), artificial intestinal fluid, bile(0.3-3.0 g/L). The results indicated that the recombinant strain survived well in artificial gastric fluids at pH 2.5-4.5 in 5 h. The recombinant Lactobacillus casei 525 could slowly grow in the artificial intestinal fluid for different time, and could survive in 0.3% bile.

Keyhole limpet hemocyanin induced Th1/Th2 imbalance in splenocytes of Balb/C mice / 中国组织工程研究

BACKGROUND:Establishing a characteristic.stable and repeatable model of Th1/Th2 imbalance in animals,is the key of studying the mechanism of Th1/Th2 imbalance.OBJECTIVE:To observe the characteristics of Th1/Th2 imbalance in splenocytes derived from Balb/C mice immnnized by keyhole limpet hemocyanin(KLH).DESIGN:A randomized control exploratory experiment.SETTING:Hebei Provincial Forensic Laboratory.Institute of Basic Medicine,Hebei Medical University.MATERLALS:The experiment was carried out in the Hebei Provincial Forensic Laboratory,Institute of Basic Medicine,Hebei Medical University from September 2005 to January 2007.Balb/C mice were adopted in this study.and all the disposals were in accordance with the guidance of animal ethics.METHODS:Balb/C mice were immunized with KLH emulsified in complete Freund's adjuvant(CFA),splenocytes were acquired,and the peak of cytokine secretion was determined in 3 groups:KLH groups of 6.25 mg,kg.12.5 mg,kg and 25 mg/kg.According to the immunizing dose and immunizing frequency.mice were divided into 7 groups:KLH groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,secondary immunity groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,as well as control group.According to the determined levels of IgG1 and IgG2a in blood serum.mice were divided into KLH group of 6.25 mg/kg and control group.MAIN OUTCOME MEASURES:Mice splenocytes supematant was detected with enzyme linked immunosorbent assay (ELISA)for the production of Th1 cytokines interferon (IFN)-γ,interleukin(IL)-2.IL-12 p40 and Th2 cytokines IL-4 and IL-5.The levels of Th1 antibody IgG2a and Th2 antibody IgG1 in blood serum were also detected by ELISA.RESULTS:The spleen derived from KLH-immunized mice appeared hypertrophy,and the number of splenocytes was manifold.Splenocytes restimulated with KLH in vitro produced much more IFN-γ,IL-2,IL-4,IL-5,but not IL-12p40.IL-2 secretion was obviously elevated after incubated for 24 hours and achieved pinnacle at 48 hours;productions of IL-4,IL-5 and IFN-γ were elevated after 24 hours,and increased gradually to 96 hours;IL-12p40 production was very low at every time point.Using different doses of KLH inlmunity once or twice,could all lead to the elevated productions of IL-2,IL-4,IL-5,IFN-γ,and the elevation of IL-4/IFN-γ ratio,but the secondary immunity group of 6.25 mg/kg KLH showed obviously higher levels than other groups(P＜0.01).The level of KLH specific antibody IgG2a and IgG1,especially IgG1 was elevated in blood serum of KLH-immunized mice.CONCLUSION:Balb/C mice immunizad with KLH emulsified in CFA can indce a Th2 predominant imbalance in splenocytes.

Analysis of current status of diagnosis, treatment and referral of pulmonary tuberculosis in general hospital / 中华全科医师杂志

Objective To investigate current status of diagnosis, treatment and referral of pulmonary tuberculosis in general hospital. Methods Statistical reports of hospital work, notification forms of tuberculosis (TB) cases on the internet, registration book of laboratory sputum mgcobacteria examinations, statistical reports of X-ray examinations for outpatient department, referral forms for mycobacteria patients, medical records for hospitalized patients at the respiratory ward of the People's Hospital, Peking University during August 2005 to July 2006 were reviewed and analyzed. Results In outpatient pulmonary department, there were 45 055 visitors during the period of August 2005 to July 2006,and 4960 of them (11.0%) had their chest X-ray examined, 1 512 (3.4%) had sputum mycobacteria examined with smear-positive in 24 cases, 189 (0.4%) were referred to specialized TB dispensaries for further diagnosis and 183 with notification forms with a notification rate of 96.8%, and 30 (0.1%) were finally diagnosed as TB after hospitalization with an interval less than 14 days between onset of symptoms and diagnosis in 27 of the 30 (90.0%) and 26 were referred to a specialized TB dispensary (86.7%) for treatment. Conclusions On general, detection, diagnosis and referral of TB in People's Hospital of Peking University are in a good situation.However,more aUenfion should be paid to the following aspects：① quantity and quality of sputum myeobacteria examinations for outpatients should be improved further,②chest radiograph reading by radiologists and clinicians need to be improved,and ③consciousness of early detection for TB need to be enforced in physicians by bronchoscopy and pathological diagnosis,especially in those with complicated and difficultly diagnosed smear-negative TB.

Chemosensitivity in vitro and its correlation with clinico-pathological parameters of patients with breast cancer / 肿瘤研究与临床

Objective To detect any association between chemosensitivity of tumor in vitro and clinico-pathological characteristics of patients with breast cancer, such as age, pathological type, pathological grade, as well as local lymph nodal status. Methods Sixty patients with breast cancer were tested with tissue culture-end point staining-computer image analysis (TECIA) after resection of the primary lesion or biopsy. Their chemosensitivity to seven common anticancer drugs were evaluated, which were fluorouracil (5-Fu), vinorelbine(NVB), cisplatin(DDP), paclitaxel(TAX), docetaxel(TAT), doxorubicin(ADM) and epirubicin(EADM). Results 31.67% of the total 60 specimens were resistant to all of 7 drugs. The sensitive rate of the tissues to a specific drugs was 60.00 % (36/60) for EADM, 54.00 %(32/60) for NVB, 36.00 %(22/60) for DDP or TAT, 30.00 %(18/60) for TAX, 26.00 %(16/60) for 5-Fu and 16.00 %(10/60) for ADM in turn. The sensitive rate of invasive carcinoma group was higher than non-invasive carcinoma group (74.47 % vs 46.15 %, P< 0.05), and grade Ⅲ group higher than grade Ⅰ or grade Ⅱ group (90.91% vs 54.55 %, 90.91% vs 55.56%, P<0.01). The differences of sensitive rates among three different age groups and between two groups with/ without lymph nodal metastasis were not significant statistically (P > 0.05). Conclusion The total sensentive rate of breast cancer might be relatively high to EADM, NVB and TAT, and TECIA could be an important reference for clinical individual chemotherapy.

Cholecystokinin octapeptide modulates T-lymphocyte subsets in KLH-immunized mice / 中国病理生理杂志

AIM: To study the effects of cholecystokinin octapeptide (CCK-8) on T-lymphocyte subsets in keyhole limpet haemocyanin (KLH)-immunized mice.METHODS: Female BALB/c mice were immunized with KLH and injected with different dosages of CCK-8 or saline simultaneously. Positive CD4+and CD8+ T cells in peripheral blood or splenocytes were measured by flow cytometry. The production and mRNA level of Th1 cytokine, IFN-? and Th2 cytokine, IL-4 in the splenocytes were evaluated by ELISA and RT-PCR, respectively. In addition, lung tissue sections were made with HE staining.RESULTS: CCK-8 downregulated the positive CD4+and CD8+ T lymphocytes both in peripheral blood and in splenocytes in KLH-immunized mice, resulting in the reduction of CD4+/CD8+ ratio.CCK-8 improved the production of IFN-?, elevated IFN-? gene transcription, whereas cut down the production of IL-4, decreased IL-4 gene transcription. CCK-8 lightened the pulmonary inflammation induced by KLH.CONCLUSION: CCK-8 inhibits CD4+T lymphocytes activation, Th2 cytokine mRNA expression and protein production in KLH-immunized mice, indicating that CCK-8 modulates adaptive immune response.