ObjectiveTo establish the quality standards for Sennae Folium（Cassia angustifolia） dispensing granules based on standard decoctions. MethodsHigh performance liquid chromatography（HPLC） specific chromatograms were established for 15 batches of Sennae Folium（C. angustifolia） standard decoctions and 10 of Sennae Folium（C. angustifolia） dispensing granules from different manufacturers， and the similarity evaluation， hierarchical cluster analysis（HCA） and principal component analysis（PCA） were performed. Linear calibration with two reference substances（LCTRS） and quantitative analysis of multi-components by single-marker（QAMS） were established for the common peaks in the specific chromatograms to determine the contents of main components in the decoction pieces， standard decoctions and dispensing granules， and to calculate their transfer rates from decoction pieces to standard decoctions and dispensing granules. ResultsThe similarities of specific chromatograms of 15 batches of Sennae Folium（C. angustifolia） standard decoctions and 10 batches of Sennae Folium（C. angustifolia） dispensing granules were all greater than 0.95， and a total of 8 characteristic peaks were calibrated， and five of them were identified， including kaempferol-3，7-O-diglucoside， apigenin-6，8-di-C-glucoside， quercetin-3-O-gentianoside， sennoside B and sennoside A. HCA and PCA results showed that there were certain differences in the composition of different batches of standard decoctions， but no clustering was observed in the production area. As the standard decoctions， the extract rate of 15 batches of samples was 26.54%-45.38%， the contents of kaempferol-3，7-O-diglucoside， apigenin-6，8-di-C-glucoside， quercetin-3-O-gentianoside， sennoside B and sennoside A were 12.16-19.26， 2.57-4.94， 3.27-5.11， 6.75-11.39， 4.69-7.79 mg·g^-1， and their transfer rates from decoction pieces to standard decoctions were 45.41%-79.02%， 29.12%-55.07%， 40.52%-67.90%， 24.72%-49.12%， 27.54%-49.34%， respectively. The extract rates of Sennae Folium（C. angustifolia） dispensing granules（C8-C10） were 38.10%-39.50%， the transfer rates of the above five components from decoction pieces to dispensing granules were 72.85%-73.58%， 53.43%-53.94%， 40.19%-40.74%， 24.62%-25.00%， 28.65%-29.11%， respectively， which were generally consistent with the transfer rates from decoction pieces to standard decoctions. ConclusionCompared with the relative retention time method， LCTRS has higher prediction accuracy and is more suitable for chromatographic columns. The established quality control standard of Sennae Folium（C. angustifolia） dispensing granules based on standard decoction is reasonable and reliable， and all indicators of samples from different manufacturers are within the range specified based on the standard decoction， which can provide reference for the quality control and process research of this dispensing granules.

ObjectiveTo establish the specific chromatogram and quantitative analysis of multi-components by single-marker（QAMS） based on linear calibration using two reference substances（LCTRS）， explore the consistency between Hedyotis Herba dispensing granules and standard decoction， and evaluate the quality of the dispensing granules. MethodsHigh performance liquid chromatography（HPLC） specific chromatogram was established based on 15 batches of Hedyotis Herba standard decoction and 10 batches of the dispensing granules， and LCTRS was used to locate chromatographic peaks. The actual retention times of 7 characteristic peaks in the specific chromatogram was measured on 24 different types of C₁₈ columns， taking deacetyl asperulosidic acid and asperulosidic acid as the dual standard compounds， the retention times of the other 5 characteristic peaks were predicted and validated. Based on this， QAMS was developed to determine the contents of four components（deacetyl asperulosidic acid， deacetyl asperulosidic acid methyl ester， asperulosidic acid， and p-coumaric acid）. Then， the relative correction factors of deacetyl asperulosidic acid， deacetyl asperulosidic acid methyl ester and p-coumaric acid were calculated using the reference peak of asperulosidic acid in the dual standard compounds， and each component was quantified accordingly. Finally， the consistency between the dispensing granules and standard decoction was assessed by taking extract rate of the standard decoction， consistency of the specific chromatograms， contents and transfer rates of the indicator components as indexes， and the quality of the dispensing granules was evaluated. ResultsThere were 7 common peaks in the characteristic chromatogram of samples of Hedyotis Herba standard decoction and the dispensing granules， and four of them were identified by reference standards， namely deacetyl asperulosidic acid（peak 1）， deacetyl asperulosidic acid methyl ester（peak 3）， asperulosidic acid（peak 6） and p-coumaric acid（peak 7）. The similarity between the dispensing granules and the standard decoction was >0.9. The absolute deviation in the predicted retention time for each component by LCTRS was lower than that of the relative retention time method. The extract rate of the 15 batches of Hedyotis Herba standard decoction ranged from 7.89% to 14.60%， the contents of deacetyl asperulosidic acid， deacetyl asperulosidic acid methyl ester， asperulosidic acid and p-coumaric acid were 6.62-19.70， 3.83-17.99， 1.57-6.69， 1.62-4.52 mg·g^-1， and the transfer rates of these components from decoction pieces to the standard decoction were 22.89%-39.60%， 34.03%-62.24%， 24.25%-43.70%， and 40.58%-73.71%， respectively. The extract rate， index component contents and transfer rates from decoction pieces to the three batches of Hedyotis Herba dispensing granules（P1-P3）， produced by manufacturer A， were similar to those of the standard decoction prepared from the same batch of decoction pieces， and all fell within the specified range. The contents of the 4 indicator components in 7 batches of the dispensing granules（P4-P10） from manufacturers B-E were all within the range of the content converted from the standard decoction based on the quantity of the dispensing granules. ConclusionThe established specific chromatogram and QAMS based on LCTRS are reasonable and reliable. Based on the evaluation indicators of standard decoction yield， consistency of specific chromatograms， contents and transfer rates of the four index components， the 10 batches of Hedyotis Herba dispensing granules from various manufacturers have exhibited good consistency with the standard decoction， indicating that the current production process is relatively reasonable.

ObjectiveTo establish the specific chromatogram of Chuanxiong Rhizoma dispensing granules（CRdg）， and to evaluate its quality by chemometrics and two reference substances for determination of multiple components（TRSDMC）. MethodsHigh performance liquid chromatography（HPLC） specific chromatograms were established using 13 batches of CRdg from 7 manufacturers， and preliminary quality evaluation was performed by similarity evaluation and chemometrics analysis. Eight characteristic peaks in the specific chromatogram of CRdg were measured on 22 different types of C₁₈ columns， and the actual retention times were recorded. Taking chlorogenic acid（peak 1） and senkyunolide A（peak 8） as double standard compounds， the retention times of the eight characteristic peaks were predicted by linear calibration using two reference substances（LCTRS）， and the method was validated on three other columns of different brands. Taking chlorogenic acid as reference peak， the relative correction factor method（RCFM） was used to quantify cryptochlorogenic acid， caffeic acid， ferulic acid， senkyunolide I and senkyunolide A， and the results were compared with the external standard method（ESM）. ResultsThe similarities of specific chromatograms of 13 batches of CRdg were all >0.90， and a total of 8 characteristic peaks were calibrated， and six of them were identified， including chlorogenic acid（peak 1）， cryptochlorogenic acid（peak 2）， caffeic acid（peak 3）， ferulic acid（peak 5）， senkyunolide I（peak 6） and senkyunolide A（peak 8）. Through chemometric analysis， it was found that ferulic acid， chlorogenic acid， senkyunolide I and cryptochlorogenic acid were the main components causing quality difference in CRdg， and the accuracy of LCTRS in predicting the retention time of 8 characteristic peaks was superior to that of the relative retention time method（RRT）. Further comparison of the results obtained from RCFM and ESM showed that there was no statistically significant difference between the two methods. ConclusionA quality evaluation method for CRdg based on HPLC specific chromatogram and TRSDMC is established， its qualitative accuracy is better than that of RRT， the quantitative accuracy is similar to that of ESM， and 4 quality-differentiated components among different manufacturers are found. This method is stable and reliable， and has reference value for the quality evaluation of other dispensing granules.

ObjectiveTo establish the specific chromatogram of Chuanxiong Rhizoma dispensing granules（CRdg）， and to evaluate its quality by chemometrics and two reference substances for determination of multiple components（TRSDMC）. MethodsHigh performance liquid chromatography（HPLC） specific chromatograms were established using 13 batches of CRdg from 7 manufacturers， and preliminary quality evaluation was performed by similarity evaluation and chemometrics analysis. Eight characteristic peaks in the specific chromatogram of CRdg were measured on 22 different types of C₁₈ columns， and the actual retention times were recorded. Taking chlorogenic acid（peak 1） and senkyunolide A（peak 8） as double standard compounds， the retention times of the eight characteristic peaks were predicted by linear calibration using two reference substances（LCTRS）， and the method was validated on three other columns of different brands. Taking chlorogenic acid as reference peak， the relative correction factor method（RCFM） was used to quantify cryptochlorogenic acid， caffeic acid， ferulic acid， senkyunolide I and senkyunolide A， and the results were compared with the external standard method（ESM）. ResultsThe similarities of specific chromatograms of 13 batches of CRdg were all >0.90， and a total of 8 characteristic peaks were calibrated， and six of them were identified， including chlorogenic acid（peak 1）， cryptochlorogenic acid（peak 2）， caffeic acid（peak 3）， ferulic acid（peak 5）， senkyunolide I（peak 6） and senkyunolide A（peak 8）. Through chemometric analysis， it was found that ferulic acid， chlorogenic acid， senkyunolide I and cryptochlorogenic acid were the main components causing quality difference in CRdg， and the accuracy of LCTRS in predicting the retention time of 8 characteristic peaks was superior to that of the relative retention time method（RRT）. Further comparison of the results obtained from RCFM and ESM showed that there was no statistically significant difference between the two methods. ConclusionA quality evaluation method for CRdg based on HPLC specific chromatogram and TRSDMC is established， its qualitative accuracy is better than that of RRT， the quantitative accuracy is similar to that of ESM， and 4 quality-differentiated components among different manufacturers are found. This method is stable and reliable， and has reference value for the quality evaluation of other dispensing granules.

ObjectiveTo establish the quality standard for Fraxini Cortex（Fraxinus chinensis） dispensing granules based on standard decoction， and to provide a basis for the quality control of this dispensing granules. MethodHigh performance liquid chromatography（HPLC） specific chromatograms of 15 batches of Fraxini Cortex（F. chinensis） standard decoctions and 3 batches of Fraxini Cortex（F. chinensis） dispensing granules were established with the mobile phase of 0.1% phosphoric acid aqueous solution（A）-acetonitrile（B） for gradient elution（0-10 min， 12%-15%B； 10-30 min， 15%-32%B） and the detection wavelength of 220 nm. And similarity evaluation， cluster analysis and principal component analysis（PCA） were also carried out. HPLC quantitative analysis of multi-components by single marker（QAMS） was established to determine the contents of the main components in the standard decoctions and dispensing granules. The contents of the corresponding components in Fraxini Cortex（F. chinensis） decoction pieces were also detected， and the transfer rates from decoction pieces to standard decoctions and dispensing granules were calculated. ResultThe similarities between specific chromatograms of 15 batches of Fraxini Cortex（F. chinensis） standard decoctions and 3 batches of Fraxini Cortex（F. chinensis） dispensing granules were all>0.9， and 7 common peaks were identified. The results of cluster analysis and PCA showed that there was some differences in the composition of different batches of standard decoctions， but did not show aggregation of origin. As the standard decoctions， the extract rate was 6.18%-11.62%， the contents of esculin， syringin， fraxin， esculetin， fraxetin， calceolarioside B were 44.92-103.51， 1.36-11.87， 33.26-90.73， 4.63-29.75， 2.40-16.86， 2.49-17.35 mg·g^-1， and the transfer rates from decoction pieces to standard decoction were 25.21%-42.54%， 52.57%-88.84%， 43.43%-79.45%， 49.15%-88.27%， 49.22%-72.69%， 27.66%-47.67%， respectively. The extract rates of Fraxini Cortex（F. chinensis） dispensing granules were 10.4%-10.7%， the transfer rates of the above six components from decoction pieces to dispensing granules were 42.76%-43.17%， 80.01%-80.90%， 59.59%-59.88%， 51.35%-52.67%， 60.50%-60.93%， 37.98%-38.37%， respectively， which were generally consistent with the transfer rates from decoction pieces to standard decoctions. ConclusionThe established quality control standard of Fraxini Cortex（F. chinensis） dispensing granules based on standard decoctions is reasonable and reliable， which can provide reference for the quality control and process research of this dispensing granules.

Objective To observe the effects of Soyasaponins on inflammatory factors, antioxidant activity and exercise ability in rats with severe heat stroke. Methods Eighty male Sprague-Dawley (SD) rats were randomly divided into normal control group, heat shock model group, saline control group and Soyasaponin group, The rats that died during the experiment or with a low rectal temperature (< 41℃) were excluded, and finally 54 rats were included, 18 rats remaining in each group. The rats in the heat shock model group were placed in the simulated hot climate animal cabin at 30 ℃, and the temperature within 30 minutes was raised to 39 ℃ in the cabin with 65% humidity; in the mean time, the rat models of heat shock were replicated under the following situations: let the rats exercise on a treadmill with running speed set at 15 m/min, slope degree 0°, once running for 8 minutes, interval 2 minutes and the heat shock time was 90 minutes, the rats in the normal control group were fed in an environment with temperature ranging from 23-25 ℃ and relative humidity ranging from 50%-70%. After the establishment of models, the saline control group and Soyasaponin group were given daily saline and Soyasaponin (10 mg/kg) respectively by gavage for 3 consecutive months, while the heat shock model group was not given any treatment. The femoral artery blood was collected 24 hours after the rats left the cabin. The serum levels of interleukins (IL-6, IL-1β), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), malonaldehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) were measured by enzyme-linked immunosorbent (ELISA) and the contents of serum hemoglobin (Hb), serum urea (BUN), lactate dehydrogenase (LDH) and blood lactic acid (Lac) were measured by automatie biochemical analyzer. Results The levels of IL-6, IL-1β, TNF-α, IFN-γ, MDA, Hb, BUN, LDH, Lac in heat shock model group were significantly higher than those of the normal control group [IL-6 (ng/L): 86.17±4.82 vs. 12.60±3.49, IL-1β (ng/L): 83.00±5.98 vs. 15.70±3.64, TNF-α (ng/L): 72.22±6.93 vs. 13.75±2.69, IFN-γ (ng/L): 36.22±3.02 vs. 7.35±1.60, MDA (nmol/mg): 19.78±4.56 vs. 6.40±1.35, Hb (g/L): 136.22±1.93 vs. 126.75±5.84, BUN (mmol/L):21.06±3.44 vs. 5.65±1.35, LDH (μmoL·s-1·L-1): 9.65±0.83 vs. 2.12±0.17, Lac (mmol/L): 552.56±78.33 vs. 1.32±0.18, all P ＜ 0.05], SOD and GSH-Px were significantly lower than those in normal control group [SOD (kU/L):97.89±10.57 vs. 126.65±11.35, GSH-Px (kU/L): 19.22±2.58 vs. 43.45±4.02]; however, the levels of IL-6, IL-1β, TNF-α, IFN-γ, MDA, BUN, LDH and Lac in Soyasaponin group were significantly lower than those in heat shock model group [IL-6 (ng/L): 45.28±3.54 vs. 86.17±4.82, IL-1β (ng/L): 41.61±2.93 vs. 83.00±5.98, TNF-α (ng/L):37.22±2.46 vs. 72.22±6.93, IFN-γ (ng/L): 19.22±2.60 vs. 36.22±3.02, MDA (nmol/mg): 11.28±1.74 vs. 19.78±4.56, BUN (mmol/L): 11.78±2.13 vs. 21.06±3.44, LDH (μmoL·s-1·L-1): 3.70±0.26 vs. 9.65±0.83, Lac (mmol/L): 274.56±59.08 vs. 552.56±78.33, all P < 0.01], SOD, GSH-Px and Hb were significantly higher than those of heat shock model group [SOD (kU/L): 116.11±11.28 vs. 97.89±10.57, GSH-Px (kU/L): 31.17±2.90 vs. 19.22±2.58, Hb (g/L): 141.33±3.79 vs. 136.22±1.93, all P < 0.01]; there were no significant statistical differences in above indexes between heat shock model group and saline control group (all P > 0.05). Conclusion After heat shock and exercise management, the production and release of inflammatory factors are increased, and the level of lipid peroxidation was elevated in rats. The Soyasaponin can improve the ability to withstand heat shock and strong exercise by reducing the production and release of inflammatory factors and lipid peroxidation in the rats with severe heatstroke.

Bilateral mandibular first molars of 36 rabbits were extracted.The tooth extraction wounds were treated by:platelet-rich fibrin +acellular dermal matrix(group A),platelet-rich fibrin(group B),acellular dermal matrix (group C) and without treatment (group D,the control) (n =9) respectively.The measurments of alveolar bone height and width showed that there were no significant differences among groups at different times(P ＞ 0.05).Bone histomorphometry showed that at the 2th and 4th week,the best result was found in group A(P＜0.01).While,at the 8th week,the result of group A was still better than that of other 3 ones (P ＜ 0.01),but group B and C showed no significant difference(P ＞ 0.05).The combination of PRF and ADM shows the most significant effect.

PURPOSE: This study aimed to determine the oncologic efficacy of gonadotropin-releasing hormone (GnRH) agonist treatment concurrent with chemotherapy in a neoadjuvant setting. METHODS: A retrospective analysis was performed on 332 cases of invasive breast cancer in patients who were <40 years old at diagnosis and received GnRH agonists concurrent with neoadjuvant chemotherapy (GnRH agonist group) or neoadjuvant chemotherapy alone (neochemotherapy-alone group) from December 2010 to September 2014. Pathologic complete response rates (pCR) and Ki-67 changes were evaluated between the two groups. RESULTS: Median age was 32+/-3.9 and 36+/-3.0 years in the GnRH agonist group and neochemotherapy-alone group, respectively (p<0.001). After adjustment for tumor size, grade, lymph node metastasis, hormone receptor (HR) status, and chemotherapy regimen, the GnRH agonist group exhibited a higher pCR rate with an odds ratio (OR) of 2.98 (95% confidence interval [CI], 1.37-6.34) and a greater decrease in Ki-67 expression after treatment (p=0.05) than the neochemotherapy-alone group. For HR-negative tumors, the GnRH agonist group showed a higher pCR rate (multivariate OR, 3.50; 95% CI, 1.37-8.95) and a greater decrease in Ki-67 expression (p=0.047). For HR-positive breast cancer, the pCR rate, change in Ki-67 index, and clinical response were higher, and preoperative endocrine prognostic index scores were lower, in the GnRH agonist group, but these did not reach statistical significance. CONCLUSION: Concurrent administration of GnRH agonists during neoadjuvant chemotherapy improved pCR rates and suppressed Ki-67 expression, especially in HR-negative tumors.
Breast Neoplasms* ; Breast* ; Diagnosis ; Drug Therapy* ; Gonadotropin-Releasing Hormone* ; Humans ; Lymph Nodes ; Neoplasm Metastasis ; Odds Ratio ; Polymerase Chain Reaction ; Retrospective Studies

BACKGROUND:Cytokines and neurotrophic factors secreted from human umbilical cord blood-derived mesenchymal stem cells secrete have neuroprotective effects on cerebral ischemia-reperfusion injury, but there are few reports about intranasal administration of human umbilical cord blood-derived mesenchymal stem cellconditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the protective effects of intranasal administration of human umbilical cord-derived mesenchymal stem cells-conditioned medium on neurologic function of rats with cerebral ischemia-reperfusion injury.
METHODS:Adult rats were subjected to 2 hours of right middle cerebral artery occlusion and the human umbilical cord-derived mesenchymal stem cells were isolated from the postpartum human cord. We made the conditioned medium of human umbilical cord-derived mesenchymal stem cells. Ischemic rats were randomized and assigned to three groups and were treated by intranasal routine starting 24 hours after middle cerebral artery occlusion with:(1) saline for control group;(2) Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium for medium control group;(3) conditioned medium treatment group (10mL/kg) daily for 14 days. Behavioral tests (foot fault test, and modified Neurological Severity Score) were performed before and at 1, 7, 14 days after middle cerebral artery occlusion.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperatively 1 day (P>0.05). Compared to the control and medium control group rats, respectively, rats in the conditioned medium group significantly improved functional outcome after stroke in days 7 and 14 (P<0.05). There was also no significant difference in functional tests between the control group and medium control group in days 7 and 14 (P>0.05). These results suggest that human umbilical cord-derived mesenchymal stem cells-conditioned medium via intranasal administration can significantly improve neurologic functional outcome after cerebral ischemia-reperfusion injury.

ObjectiveTo establish a novel assay for HIV-1 p24 ultrasensitive detection based on Gold Nanoparticle Probe (GNP) and PCR.MethodsSandwich ELISA method was established by a pair of anti-p24 monoclonal antibodies (mAbs),1G12 and 1D4,and was used to detect recombinant HIV-1 p24 antigen.The bio-barcode DNA was 47 bp,selected from genome of Arabidopsis,and formed double-stranded DNA by hybridization with the capture DNA (complementary with bio-barcode DNA) modified with sulfhydryl.Then double-stranded DNA were conjugated on the surface of 1D4-modified gold nanoparticles by sulfhydryl,and the Gold Nanoparticle Probe was produced.1G12 was precoated in the micropaltes,and in the presence of target recombinant HIV-1 p24 protein,a sandwich immuno-complex would form by adding GNP.Then the bio-barcode DNA in the immuno-complex were released by heating as detection signal,and consequently characterized by the polymerase chain reaction (PCR) with synthesized special primers and analyzed by 4％ agar gel electrophoresis,so HIV-1 p24 antigen could be evaluated.The sensitivity comparison between the new assay and ELISA can be done.ResultsSandwich ELISA was used to quantify HIV-1 p24 antigen by monoclonal antibodies 1G12 and 1D4,and the limit of detection (LOD) was 1000 pg/ml.The new GNP assay was established by the same pair of antibodies,combined with PCR and agar gel electrophoresis,and was used to indirectly detect HIV-1 p24 antigen.The band intensity of PCR products paralleled with the quantity of HIV-1 p24 antigen,and the limit of detection (LOD) could reach down to 1 pg/ml.ConclusionThe new assay based on GNP and PCR was efficient in the detection of HIV-1 p24,which is at least 3 orders of magnitude more sensitive than traditional ELISA.