Experimental studies on the repair and restitution of cartilage by cartilage acellular extracellular matrix and adipose tissue-derived stem cells / 中华耳鼻咽喉头颈外科杂志

Objective@#To investigate the effects of the repair and restitution of ear-shaped cartilage by adipose tissue-derived stem cells(ADSC) and cartilage acellular extracellular matrix.@*Methods@#ADSC were extracted by digesting with collagenase type II from the adipose tissue from 32 patients with adiposity whose fats were drawn, and were cultured and subcultured in vitro. The natural biological scaffolds were prepared by acellular method using porcine ear cartilage, and then the second generation ADSC(5.0×107/ml) were inoculated on the preformed natural bio-scaffold scaffold by culturing in vitro for 3 days to form a cell scaffold complex. 32 New Zealand white rabbits were randomly divided into the experimental groups, the control group A, the control group B and the control group C. All New Zealand white rabbits were modeled by ear cartilage defects. The cell scaffolds composite was implanted into the experimental group of the ear cartilage defects of rabbits, the ADSC were implanted into the control group A, the cartilage acellular extracellular matrix scaffold was implanted into the control group B and the control group C was modeled only by ear cartilage defects. After 16 weeks, the animals were sacrificed and the repair effect was observed by gross appearance and histological examinations including HE, Toluidine blue staining, Safranin O and typeⅡ collagen staining. Its were quantitatively analyzed by positive staining results of type Ⅱ collagen. Ear cartilage tissue elasticity was detected. SPSS 17.0 software was used to analyze the data.@*Results@#The cartilage defects in the experimental group were repaired well by general shape observation and those in the control group was filled in with granulation tissue. There were significant differences between the experimental group and the control group in the wet weight(P<0.05). HE staining showed that cartilage cavities formed in articular cartilage defects, and only the fibrous tissue was filled with the ear cartilage defect in the control groups. In the repair area, Toluidine blue staining, Safranin O and type Ⅱ collagen staining were positive in the experimental group, and negative in the control groups. There was no significant difference between the experimental group and the normal ear cartilage in the ear cartilage elastic constant detection(P>0.05).@*Conclusions@#The mechanics and histology of rabbit ear neonatal cartilage constructed by ADSC combined with cartilage acellular matrix are close to normal ear cartilage. Cartilage acellular matrix material combined with adipose-derived stem cells has good repair and reconstruction ability for ear cartilage defects, which possesses potential clinical application value.

Analysis and adscription of volatiles fromGuizhi Tang using gas chromatography-mass spectroscopy and improvement of the learning and memory in mice / 国际中医中药杂志

Objective Analysis and adscription of volatiles from Guizhi Tang and study on its improvement of the learning and memory in dementia mice induced by scopolamine.Methods The volatile oil from Guizhi Tang(GZT),Guizhi and Shengjiang was extracted using steam distillation method and was analyzed by GC-MS. Morris water maze and step-down test were carried out for obtain the difference of the learning and memory improvement in 40 ICR mice from randomized groups, such as the control group, the model group, the donepezil group (2 mg/kg), the low dose of volatile oil of GZT (5 mg/kg), and the high dose of volatile oil of GZT (20 mg/kg), and ACh, AchE, BchE and chE in serum were detected by ELISA. Results Among 38 identyfied volatile ingredients from GZT, 18(44% in weight) was from Guizhi, and 9 was from Shengjiang. Compared with the model group, the low and high dose of GZT volatile oil significantly increased swimming distance ratio in destination quadrant (26.74% ± 16.42%vs.9.42% ± 8.50%, P<0.05); goal quadrant time scale (43.51% ± 25.12%vs. 14.50% ± 12.23%,P<0.05)) increased significantly than the model group ; the number of errors in the experiment platform (1.63 ± 1.19vs. 0.25 ± 0.46, P<0.05) obviously increased than model group ; platform test in the made errors times (0.57 ± 0.98vs. 4.43 ± 2.4, P<0.05) significantly reduced. The GZT total volatile oil groups significantly reduced cognitive obstacles small rat serum in the cholinester enzyme (chE) (140.90 ± 3.27, 144.79 ± 6.71vs. 134.49 ± 3.36,P<0.05); acetylcholinesterase (AchE) (3.30 ± 1.31, 3.94 ± 0.78 vs.8.52 ± 3.39,P<0.05); butyrylcholinesterase (BchE) (3.22 ± 0.45, 3.66 ± 0.53vs. 7.99 ± 0.79,P<0.05); and acetylcholine (Ach) (4.10 ± 0.38, 3.03 ± 0.25vs.1.72 ± 0.50, P<0.05) significantly increased.Conclusions The GZT volatile oil mainly from Guizhi and Shengjiang can improve the learning and memory ability in dementia mice induced by scopolamine via a cholinergic mechnism.

Study on the Preparation Technology of Fanhuncao Sustained-release Dropping Pills / 中国药房

OBJECTIVE:To study the preparation technology of Fanhuncao sustained-release dropping pills. METHODS:The preparation technology of Fanhuncao sustained-release dropping pills was optimized by orthogonal design and single factor test with PEG4000,PEG6000 and glycerol monostearate(GM)as carrier materials,using sphericity,pill weight difference and appearance quality as index,GM-PEG weight ratio,liquid temperature,nozzle temperature,dipping speed,dropping distance,diameter of emitter as factors and the verification test was conducted. RESULTS:The optimal technology was that the ratio of GM-PEG was 3∶7;the temperature of drug mixture was 85 ℃;the dropping speed was 40 drop/min;dropping distance was 4 cm;and the conden-sate temperature was 15 ℃;diameter of emitter was 4 mm. RSD of each index of 3 batches of dripping pills were all ≤0.58%(n=3)in verification test;the average content of chlorogenic acid was 0.14 mg/g;the average pill weight difference was 3.21%;the average sphericity was 9.43;and the average appearance quality was 4.33. Q1 h were 23.4%,24.4% and 23.3% in average(n=6),and Q12 h were 89.6%,91.2% and 91.5%(n=6). CONCLUSIONS:The optimal preparation technology is stable and simple, and can be used for industrial production of Fanhuncao sustained-release dropping pills.

Establishment and application of method to screen anti-EV71 positive plasma / 中华微生物学和免疫学杂志

Objective To evaluate the titers of EV71 neutralizing antibody in donators' plasma,and to establish a high throughput screening method to screening anti-EV71 positive plasma for the production of EV7l-specific IVIG in practice.Methods The EV71 neutralizing antibody titers in plasma samples and in commercial IVIGs (intravenous immunoglobulins) were evaluated by micro-cytopathic effect neutralization test (MCPENT),and a selecting criteria was determined for screening anti-EV71 positive plasma.Then,a high throughput neutralizing method for screening plasma was established and a comparison with MCPENT was conducted.EV71-specific IVIG was prepared from screened anti-EV71 source plasma,and its neutralizing activity was evaluated in cell model and mouse model.Results The results of MCPENT showed that the titers of EVT1 neutralizing antibody were 1 ∶ 50 or above in 20％ of the source plasma,and reached 1 ∶ 180 after being pooled,approaching to the titer of commercial IVIG.In 480 samples,the coincidence rate between the established high throughput neutralizing method and MCPENT was 88.1％.The quality of prepared EV71-specific IVIG was complied with the national standard,and its neutralizing potency was 16 times of that of IVIG,suggesting EV71-specific IVIG had a significantly improved protecting effect on experimental neonatal mice.Conclusion The screening method and the screening criteria for EV71 specific plasma were successfully established.The established high throughput neutralizing method that was operated easily could be used for plasma screening in the development of EV71-specific IVIG.The prepared EV71-specific IVIG,whose neutralizing potency was significantly improved,showed a markedly protective effect on experimental neonatal mice,suggesting it might be very important for the prevention and treatment of handfoot-and-mouth disease.

Cinnamaldehyde decreases interleukin-1beta induced PGE2 production by down-regulation of mPGES-1 and COX-2 expression in mouse macrophage RAW264.7 cells / 中国中药杂志

Cinnamaldehyde was shown to have significant anti-inflammatory and anti-pyretic actions in studies from both others' and our lab. Prostaglandin E2 (PGE2) plays a key role in generation of these pathological states, while PGE, synthase-1 (mPGES-1) is one of crucial biological elements in the process of PGE2 production. And as a downstream inducible terminal prostaglandin synthase of COX-2, mPGES-1 is now regarded as a more promising novel drug target than COX-2 and is attracting more and more attention from both academia and pharmaceutical industry. The purpose of present study was to further investigate the anti-inflammatory and antipyretic molecular mechanisms of cinnamaldehyde based on the mouse macrophage cell line RAW264. 7 in vitro. The PGE2 was identified by using the method of enzyme-linked immunosorbent assay (ELISA) and the expression of COX-2 and mPGES-1 at mRNA and protein levels was detected by the Real-time PCR and Western blotting methods respectively. The experimental results suggested that cinnamaldehyde could evidently reverse the increased production of PGE2induced by IL-1beta. Moreover, the up-regulated expression levels of mPGES-1 and COX-2 were significatly decreased. Together, these results provide compelling evidence that the down-regulated actions to both the production of PGE2 as well as the expression of mPGES-I might account for, at least in part, the anti-inflammatory and anti-pyretic effects of cinnamaldehyde.

Effects of ingredients from Chinese herbs with nature of cold or hot on expression of TRPV1 and TRPM8 / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of the ingredients from Chinese herbs with the nature of cold or hot on the expression of TRPV1 and TRPM8.</p><p><b>METHOD</b>The effects of ingredients from herbs on primary culture DRG neurons are observed in vitro. The expression quantity of gene is detected by the method of real time PCR. the 2 (-deltadeltaCT) method is applied to analyze the data.</p><p><b>RESULT</b>Ingredients from herbs with the nature of cold up-regulate the expression level of TRPV1 and down-regulate that of TRPM8, especially under the temperature condition of 39 degrees C; while ingredients from herbs with the nature of hot up-regulate the expression level of TRPM8 and down-regulated that of TRPV1, which is more significant under the temperature condition of 19 degrees C.</p><p><b>CONCLUSION</b>The regulatory changes of TRPV1 and TRPM8 mRNA expression induced by the chemical ingredients might be related to the cold and hot natures of the herbs from which the ingredients are extracted. And this could be one of the therapeutic mechanisms for the treatment of Chinese herbal medicines to cold- and heat-related diseases.</p>

Expression of hTERT mRNA in Anip973 and Anip973/NVB cell detected by quantitative real-time PCR / 中国肺癌杂志

<p><b>BACKGROUND AND OBJECTIVE</b>Human telomerase reverse transcriptase is the catalytic subunit of telomerase, and its activity is correlated with cell's sensitivity to chemotherapy. The aim of this study is to investigate the differential expression of human telomerase reverse transcriptase (hTERT) mRNA in human lung adenocarcinoma cell line Anip973 and Anip973/NVB, and to observe the correlation between hTERT mRNA and drug-resistance.</p><p><b>METHODS</b>The real-time fluorescence quantitative RT-PCR was used to detect the change of hTERT mRNA in human lung adenocarcinoma drug-resistant cell Anip973/NVB and parental cell Anip973 treated by NVB.</p><p><b>RESULTS</b>In the control group, the expression of hTERT mRNA showed no significant difference between drug-resistant cell Anip973/NVB and parental cell Anip973. After been treated by NVB, the expression of hTERT mRNA in parental cell was significantly decreased (P < 0.01), and drug-resistant cell Anip973/ NVB had no evidently variant (P > 0.05). The down-regulated hTERT mRNA in Anip973 cell was higher than that in Anip973/ NVB cell.</p><p><b>CONCLUSION</b>Telomerase correlates with the drug-resistant cell A973/NVB, and telomerase may be a new target for multi-drug resistant inversion.</p>

Content Determination of Oleanolic Acid and Ursolic Acid in Prunella vulgaris by HPLC / 中国药房

OBJECTIVE:To establish HPLC method for the content determination of oleanolic acid and ursolic acid in Prunella vulgris with ?-CD as additive agent of mobile phase.METHODS:The sample was Prunella vulgaris.The separation was performed on a Tigerkin ODS3(250 mm?4.6 mm,5 ?m) column with mobile phase consisted of methanol-10 mmol?L-1 ?-CD 1% ammonium acetate(86:14) at flow rate of 1.0 mL?min-1.The column temperature was set at 40 ℃ and UV detection wavelength was 210 nm.RESULTS:The linear range of oleanolic acid and ursolic acid were 0.5～5.0 ?g(r=0.999 2) and 1.0～10.0 ?g(r=0.999 8) respectively.High,medium and low average recovery of oleanolic acid were 97.3%(RSD=1.04%),98.0%(RSD=0.87%),101.6%(RSD=1.65%),respectively.Those of ursolic acid were 97.8%(RSD=1.17%),98.7%(RSD=0.93%),100.4%(RSD=1.72%),respectively.CONCLUSION:The new method is simple,accurate and reproducible for the quality control of P.vulgaris and simultaneous determination of oleanolic acid and ursolic acid.

Determination of Forsythin of Yinqiao Jiedu Pills by RP-HPLC / 中国药房

OBJECTIVE:To estabish a RP-HPLC method for the content determination of forsythin of Yinqiao jiedu pills.METHODS:The determination was performed using SHIM-PACK VP-ODS column(150 mm ? 4.6 mm,5 ?m) with column temperature at 30 ℃.The mobile phase was acetonitril-water(25∶75) with flow rate at 1.0 mL?min-1.The detective wavelength was set at 277 nm.RESULTS:The linear range of forsythin was 0.453～3.481 ?g(r=0.999 8).The mean recovery was 99.50%(RSD=1.15%,n=6).CONCLUSION:The method was simple,rapid,accurate and suitable for the quality control of Yinqiao jiedu pills.

Changes of adenyl cyclase and phosphodiesterase activities at different time point in hypothalamus of rats with fever and hypothermia / 中国病理生理杂志

AIM: To observe the changes of adengl cyclase(AC) and phosphodiesterase(PDE) activities of at different time point in hypothalamus of rats with fever and hypothermia. METHODS: Radioisotope method was used to measure the activity of AC and PDE. RESULTS:The fresh yeast caused rats fever after subcutaneous injection 4h( P

Effect of Xionggui Dripping Pills on Sensitivity of Baroreflex in Selective Sinoaortic Denervated Rats / 中国药房

OBJECTIVE:To observe the effect of Xionggui dripping pills on sensitivity of baroreflex in selective sinoaortic denervated rats.METHODS:The dynamic blood pressures were monitored in selective sinoaortic denervated rats in waking stait,and the changes of baroreflex senitivity(BRS)were measured by modified SU Smyth method.RESULTS:Xionggui drip_ ping pill could improve BRS in sinus denervated rats.CONCLUSION:Since Xionggui dripping pill rectifies autonomic nerve function,it can protect target organs from damage in cardiovascular diseases.

Changes of AC activity and content of cAMP in hypothalamus of rats with fever and hypothermia / 中国病理生理杂志

AIM: To observe the changes of AC activity and content of cAMP at different time point in hypothalamus of rats with fever and hypothermia. METHODS: Radioisotope method was used to measure the enzymatic activity of AC and the content of cAMP. RESULTS:(1)The fresh yeast caused fever after making model 4 h( P