Objective: To study the relativity between the heating degree and color value of Chinese materia medica pieces. Methods: The effects of heating temperature and time on the color value of Gardeniae Fructus and Crataegi Fructus were investigated by spectrophotometric technique. Results: The color value curve of Gardeniae Fructus after processing under the mid-and high-heating showed the better rules that the stationary phase of L* and δE* appeared in 7 min and stationary phase of a* and b* appeared in 4 min. However, the color value curve of Crataegi Fructus showed the stationary phase of L*, a*, b*, and δE* appeared in 5 min after low-heating process. After the mid-heating process, the stationary phase of L* and δE* appeared in 6 min, stationary phase of a* and b* values appeared in 5 and 8 min, respectively. After the high-heating process, the stationary phase of L* and δE* appeared in 8 min and the stationary phase of a* and b* value appeared in 5 min. Conclusion: There is the correlation of processing degree between the measurement results and the perusal observation, and the result of color measurement could provide the reference for confirming optimum parameters of processing technique.

Objective To observe the in vivo gene expression profile of the recombinant adenoassociated-2 virus mediated human GM-CSF, mouse GM-CSF (rAAV-2-hGM-CSF, rAAV-2-mGM-CSE )vector modified bone marrow mesenchymal stem cell (BMSC). Methods We transduced the BMSC by rAAV-2-hGM-CSF, rAAV-2-mGM-CSF at the condition which have acquired before respectively, then transfused the in vitro gene modified BMSC after 12 days proliferation in vitro to 6 weeks old nude mice through tail vein,while the BMSC transfused in control group hadn' t been gene modified. 2, 4, 6, 8 weeks after transfusion, count the total white blood cells and detect the hGM-CSF, mGM-CSF concentration in nude mice serum at that time point. Results Nude mice serum hGM-CSF levels were 23.77, 25.32, 19.77, 15.25 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 36.25 ng/L, the in vitro level before transfusion; mGM-CSF levels were 34.96, 34.84, 35.50, 32.93 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 25.14 ng/L, the in vitro level before transfusion; at the same time point the nude mice serum mGM-CSF levels were 17.34,17.44, 14.68, 16.85 ng/L in control group, rAAV-2-mGM-CSF transduced BMSC made the nude mice white blood cell count increased, but no changes in nude mice white blood cell count at rAAV-2-hGM-CSFtransduced BMSC and control group. Conclusion BMSC as a gene therapy vehicle, it can be gene modified in vitro, then the gene modified BMSC could let the therapeutic gene to have therapeutic effects in vivo.

Objective:To comprehend morphology changes of temporomandibular joint in adults with mandible deviation,and the correlation in these changes.Methods:The mesofault radiographs of temporomandibular joint were taken in 21 adult patients with mandible deviation.The data described morphology of temporomandibular joint were analysed.Results:In adult patients with mandible deviation,the condyle process of opposite side was anterior and inferior when compared with deflected side.The height of condyle process,the upper height of condyle process,the gradient of prosobevel of condyle process and the gradient of back bevel of glenoid fossa were augmented when compared to deflected side.The gradient of prosobevel of condyle process showed positive correlation to the prosoblank of joint and the deep of glenoid fossa,and the height of condyle process showed positive correlation to the upper height of condyle process in both sides.The gradient of back bevel of condyle process showed positive correlation to the gradient of back bevel of glenoid fossa in deflected side.The gradient of back bevel of condyle process showed negative correlation to the supper blank of joint and the height of articular tubercle in opposite side.Conclusion:There are some differences in morphology of both temporomandibular joint in adults with mandible deviation,and there is some correlation between these changes.

Objective To investigate the detection of human bocavirus (HBoV) in children with acute respiratory infection and to explore the relationship between viral load and clinical characteristics of acute respiratory infection in children.Methods A total of 4 501 nasopharyngeal secretion samples were collected from hospitalized children with acute respiratory infection from January 2013 to June 2013.HBoV-positive children were divided into simple infection group and mixed infection group.Children with HBoV DNA≥1 × 104 copy/mL were categorized into high viral load group,while those with HBoV DNA ＜1 × 104 copy/mL were categorized into low viral load group.HBoV was determined by fluorescence quantitative polymerase chain reaction (PCR).Respiratory syncytial virus (RSV),influenza virus (Inf)-A,Inf-B,parainfluenza virus (Pinf)-Ⅰ 、Pinf-Ⅱ 、Pinf-Ⅲ and adeno virus antigen were detected by direct antigen-specific immunofluorescence assays.Mycoplasm Pnuemonia was detected by real-time fluorescence quantitative PCR.Serum mycoplasma antibodies were detected by enzyme-linked immunosorbent assay (ELISA).Bacteria was detected by sputum culture.Over the same period,23 children undergoing elective inguinal hernia operation with no respiratory infection or fever were considered as control group.The percentage of peripheral blood T lymphocyte subsets were tested by flow cytometry.Inter-group differences were compared using Chi-square test or Fisher exact test.Viral loads were compared using Mann-Whitney test.Results Two hundred and twenty-two HBoV-positive cases were detected with a positive rate of 5.41％ (222/4 105),33.33％ (74/222) of which were with high viral load and 66.67％ (148/222) were with low viral load.There was a high incidence in the age group of 1-2 years.The simple HBoV infection accounted for 24.32％,including 26 cases with high viral load and 28 cases with low viral load.Wheezing was more common in patients with high viral load than those with low viral load,and the difference was statistically significant (88.46 ％ vs 42.86 ％,x2 =12.295,P=0.001).Among the 222 HBoV-positive cases,the median viral load of HBoV in simple infection group was 3.86 × 103 copy/mL,and 1.0× 103 copy/mL in mixed infection group.The difference of the viral load between these two groups was statistically significant (Z =2.906,P =0.004).Mycoplasma and Streptococcus pneumonia were most commonly detected in the 168 patients with mixed infection.Percentages of CD3+ and CD3+/CD8+ subsets were significantly lower in HBoV simple infection group and mixed infection group,compared to control group (both P＜0.05).However,percentages of CD3 /CD19+,CD19+/ CD23+ subsets were significantly higher in HBoV simple infection group and mixed infection group,compared to control group (both P＜0.05).Conclusions HBoV is one of the pathogens causing acute respiratory tract infection in children,which lead to cellular immunity dysfunction in children.Moreover,children with higher HBoV load are more likely to develop wheezing.Co-infection with other pathogens should be considered in children with low HBoV load.

Objectives To investigate the significance of electronic bronchoscopy and bronchoalveolar lavage in diagno-sis and treatment in children with pulmonary mass lesion. Methods A total of 74 hospitalized children from January 2011 to June 2012 whose imaging examinations showed massive patchy shadow were examined and treated by electronic bronchoscopy and bronchoalveolar lavage. Their clinical data were retrospectively analyzed. Results The major cause for the massive shadow was infection according to electronic bronchoscopy examination (68/74, 91.89%), 65 cases of them were lobar pneumonia, 3 cas-es were pulmonary tuberculosis followed by 5 cases of foreign body (6.76%) and one case of pneumorrhagia (1.35%).The lower left lung was the most frequently seen site of infection, followed by lower right lung. The agreement between infection sites and imaging examination was 97.30%. Bronchoalveolar lavage fluid showed that the primary pathogen of lobar pneumonia infection is Mycoplasma pneumoniae (MP) (42/65, 64.62%). The highest detection rate of MP was found in preschool group and the detec-tion rate between different age groups indicated statistically significant difference (P<0.01). The imaging examination showed pulmonary lesions in 61.54%children with lobar pneumonia improved significantly in one week. The improvement rate of pul-monary lesions was higher in infected children with short duration (1-2 weeks, 90.91%) between disease onset and electronic bronchoscopy inspection than those with longer duration (2-3 weeks, 51.72% and >3 weeks, 35.71%) (P<0.05). Conclusions Electronic bronchoscopy and bronchoalveolar lavage play dual roles in etiological diagnosis and therapy in children with pulmo-nary mass lesion.

AIM: To investigate the expression of osteopontin (OPN) and CD44 in human renal proximal tubular epithelial cells stimulated by human serum albumin (HSA). METHODS: Proximal tubular epithelial HK-2 cells were stimulated by HSA at different concentrations for different time, then OPN mRNA production was detected by RT-PCR, and OPN protein was detected by Western blotting. The expression of OPN and CD44 in HK-2 cells after stimulation for 24 h or 48 h were detected by immunofluorescence with confocal laser scanning microscope. RESULTS: Osteopontin mRNA in HK-2 cells showed a highest expression at 3 h and 48 h after HSA stimulation. However, the expression of OPN protein in HK-2 cells reached the maximum at 24 h after HSA stimulation. OPN mRNA and protein showed a strong dose-dependence relation with the concentration of HSA. HSA also stimulated HK-2 cells to express CD44 protein, the fluorescence of CD44 was most prominent at 48 h after HSA stimulation. CONCLUSION: HSA stimulates human renal proximal tubular epithelial cells to express OPN and CD44.

Objective To explore the association between rs3758539G-803A and rs10882283 T-179G polymorphism of retinol binding protein 4 (RBP4) and rosiglitazone response in Chinese type 2 diabetes mellitus (T2DM) patients.Methods A total of 472 Chinese T2DM patients and 198 healthy subjects were enrolled to identify G-803A and T-179G genotypes using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP ).assay.Forty-two T2DM patients with different G-803A or T-179G genotypes were selected to undergo a 12-week rosiglitazone treatment (4 mg/d).Serum fasting plasma glucose (FPG),postprandial plasma glucose (PPG),fasting serum insulin (FINS),glycated hemoglobin (HbAlc),postprandial serum insulin ( PINS),triglyceride (TG),low-density lipoprotein-cholesterol ( LDL-c),and high-density lipoprotein-cholesterol (HDL-c) were determined before and after the rosiglitazone treatment.Results T2DM patients with RBP4 G-803A GG genotype showed lower TG and LDL-c concentrations compared with that in the GA +AA genotype subjects.T2DM patients with RBP4 T-179G TT genotype showed lower waist-to-hip ratio (WHR),FPG and FINS values compared with that in the TG + GG genotype individuals.Patients with GG genotype of RBP4 G-803A had an enhanced rosiglitazone efficacy on FPG and FINS compared with that in the GA + AA genotype group.Patients with RBP4 T179G TG + GG genotype showed an enhanced rosiglitazone efficacy on HbAlc level compared with that in the TT genotype group.Conclusion RBP4 G-803A and T-179G polymorphism might be associated with the development of T2DM and affect the therapeutic efficacy of rosignitazone in Chinese T2DM patients.

Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.
Animals ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Electrophoresis, Polyacrylamide Gel ; Male ; Microsomes, Liver ; chemistry ; drug effects ; enzymology ; Proteomics ; Rats ; Rats, Sprague-Dawley

Objective To estimate the influence of palmitic acid (PA) on the proliferation of and production of inflammatory mediators by a human keratinocyte line HaCaT.Methods Cultured HaCaT cells were treated with PA of eight concentrations (0-200 μmol/L) for 3-24 hours followed by the evaluation of cell proliferation by using the cell counting kit-8.According to the proliferation assay,four concentrations (75,100,125,150 μmol/L) of PA were selected and used to treat HaCaT cells for 24 hours,then,fluorescence-based immunohistochemical staining was performed to observe the nuclear translocation of nuclear factor (NF)-κB p65,enzyme linked immunosorbent assay (ELISA) to determine the level of interleukin (IL)-6 in the supernatant of culture medium,real-time PCR to detect the mRNA expressions of peroxisome proliferator-activated receptor oα (PPARα) and IL-6,and Western blot to quantify the protein expressions of PPARα as well as total and nuclear NF-κB p65.Those HaCaT cells receiving no treatment served as the control group.Statistical analysis was carried out by one-factor analysis of variance using the GraphPad Prism 5.0 software.Results The HaCaT cells treated with PA of 50-175 μ mol/L showed accelerated proliferation compared with the control HaCaT cells (all P ＜ 0.05).PA from 75 to 150 μmol/L enhanced the nuclear translocation of NF-κB p65,mRNA and protein expressions of PPARα,as well as the mRNA expression and supernatant level of IL-6 in a dose-dependent manner.The relative expression level of nuclear NF-κB p65 protein was 0.4536 ± 0.0173,0.5184 ± 0.0206,0.5333 ± 0.0231,0.6160 ± 0.0297,and the supernatant level of IL-6 was (31.5677 ± 0.2268),(32.3773 ± 0.4156),(32.9837 ± 0.0029) and (33.6890 ± 0.0936) ng/L,in HaCaT cells treated with PA of 75,100,125 and 150 μmol/L,respectively,compared to 0.3237 ± 0.0114 (all P ＜ 0.01) and (30.4577 ± 0.5131) ng/L (all P ＜ 0.01) in the control HaCaT cells,respectively.Conclusions PA can accelerate the proliferation of HaCaT cells,enhance NF-κB nuclear transfer,PPARα expression and IL-6 secretion in a dose-dependent manner within a certain concentration range,and may exert a promoting role in the activation and expression of some inflammatory factors.

Objective To observe the effect of sirolimus,an autophagy enhancer,on premature senescence in fibroblasts induced by repeated exposure to a subtoxic dose of ultraviolet B (UVB).Methods Skin fibroblasts from foreskin tissue of healthy adolescents were classified into six groups:control group cultured in Dulbecco's modified Eagles' medium (DMEM) containing 1％ calf serum,UVB group receiving UVB irradiation only,sirolimus group treated with sirolimus of 10 mg/L (added after daily exchange of culture medium),and three combined groups receiving UVB irradiation immediately followed by overnight treatment with sirolimus of 0.1,1.0 and 10.0 mg/L respectively.UVB irradiation was given at a dose of 10 mJ/cm2 once a day for five successive days.After five days of treatment,cell counting kit-8 (CCK-8) was used to evaluate cell viability,β-galactosidase staining to detect senescent ceils,Western blot to quantify the expressions of p53,LC3-B and beclin 1 in these fibroblasts.Autophagy level was determined by acridine orange staining followed by fluorescence microscopy and transmission electron microscopy.Data were processed by the SPSS 16.0 software,and statistical analysis was done by one-way analysis of variance,t test and least significance difference.Results Sirolimus significantly increased the proliferative activity of fibroblasts in a dose-dependent manner,with the absorbance value at 450 nm being 0.27 ± 0.02,0.36 ± 0.04 and 0.39 ± 0.04 for fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L respectively,compared to 0.26 + 0.01 for fibroblasts irradiated with UVB only (all P ＜ 0.05).Significant differences were also observed between the fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L and those irradiated with UVB only in the percentage of β-galactosidasepositive fibroblasts (92.50％ ± 0.34％,42.40％ ± 0.53％ and 6.20％ ± 0.39％ vs.95.10％ ± 0.32％,all P ＜ 0.05)and intracellular intensity of acridine orange-induced fluorescence (36.43 ± 0.24,45.25 ± 0.33 and 48.69 ± 0.37 vs.33.99 ± 0.32,all P ＜ 0.05).Moreover,the expressions of p53,LC3-B and beclin 1 in the three combined groups differed significantly from those in the UVB group (all P ＜ 0.05).Conclusion Sirolimus can inhibit UVBinduced premature senescence likely via upregulation of autophagy in fibroblasts.