Objectives To investigate the effect of rhEPO on newborn rats with necrotizing enterocolitis (NEC). Meth-ods Sixty newborn Sprague-Dewley (SD) rats at the age of 48 hours were randomly divided into 5 groups:control group, NEC group, and intervention groups 1, 2 and 3 treated with rhEPO. The rats were fed rat breast milk substitutes and stressed under hypoxia and cold exposure to establish NEC model. The rats with NEC were treated with different doses of rhEPO (0.1U/ml, 1U/ml and 10U/ml) in intervention groups. The expression of Bcl-2 and caspase-3 were measured by immunohistochemistry, and intestinal pathological changes were observed using HE staining. The value of positive expression was analyzed by IOD (integral optical density) image analysis system. Results Abdominal distention, decreased activity and unresponsiveness occurred in NEC rats 24 hours after stress exposure, and pale skin, decreased skin temperature and respiratory rhythm change were observed in severe cases. The symptoms appeared later and milder in three intervention groups. The NEC incidence of newborn rats was as followings:control group(0%), model group(60%), intervention group 1( 30%), intervention group 2(18.2%), intervention group 3(9.1%) and the difference was signiifcant between each group (P=0.008). The grades of intestinal injury, the expression of active caspase-3 and Bcl-2 were signiifcantly different among groups (P<0.01). Intestinal injury was the most severe and the expression level of active caspase-3 was the highest in NEC group. After rhEPO treatment, the intestinal injury and the production of active caspase-3 protein were decreased, and the expression of Bcl-2 was increased. Conclusions Oral rhEPO could decrease the ex-pression of intestinal active caspase 3, and increase the expression of Bcl-2. The protective effect of rhEPO on NEC is dosede-pendent.

Objective To observe the effects of long term injection sodium salicylate on the auditory brain-stem response(ABR)and expression of glutamic acid decarboxylase -67(GAD67) in rat inferior colliculus .Methods Eighteen healthy Wistar rats were randomly divided into three groups :the sodium salicylate group (intramuscular injection of 10% sodium salicylate ,175 mg/kg ,twice daliy for 28 days) ,the saline group (intramuscular injection with saline on same does at the same time) ,the control group (without any treatment) .The rats received ABR after modeling ,then were decapitated and inferior colliculus tissues were stripped .Western blot was used to study the dif-ferent expression of GAD67 protein levels in the three groups .Results Compared with the saline group and control group ,ABR thresholds of the sodium salicylate group were significantly elevated and latency of wave Ⅲ was aslo sig-nificantly prolonged(P<0 .01) ,while there was no significant difference between the saline group and the control group(P>0 .05) .The inferior colliculus GAD67 protein expression level of sodium salicylate group was significantly higher than the saline group and control group(P<0 .01) ,while there was no significant difference between the saline group and the control group(P>0 .05) .Conclusion Long term injection of sodium salicylate can cause a change in the inferior colliculus of GAD67 protein expression and the up regulation of GAD67 expression may occur as a com-pensatory response to increase inhibiting effect .The change of GAD67 protein expression is likely as a compensatory and regulatory mechanisms for sodium salicylate ototoxicity .

The stable and controllable quality of decoction pieces is an important factor to ensure the efficacy of clinical medicine. Considering the dilemma that the existing standardization of processing mode cannot effectively eliminate the variability of quality raw ingredients, and ensure the stability between different batches, we first propose a new strategy for Chinese medicine processing technologies that coupled with individuation processed and cybernetics. In order to explain this thinking, an individual study case about different grades aconite is provided. We hope this strategy could better serve for clinical medicine, and promote the inheritance and innovation of Chinese medicine processing skills and theories.
Chemistry, Pharmaceutical ; methods ; standards ; Cybernetics ; standards ; Drug Therapy ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; toxicity ; Humans ; Quality Control

Quality control is one of the key scientific tissues in the modernization of traditional Chinese medicines (TCM). In order to overcome the deficiencies of assessment indexes, including little systematization and quantification, as well as loose association with clinical efficacy and dosage, a new integrated method named controllability pyramid ( CP) is first proposed in this paper. In addition, some study cases are used to explain how this model is constructed. We hope the establishment of CP could promote the clinical-orien- ted integrated innovation research of TCM, and provide control strategy and technology examples for improving the quality of Chinese medicines and clinical efficacy.
China ; Drug Evaluation ; methods ; standards ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; Quality Control

Good medicinal herbs, good drugs. Good evaluation method and indices are the prerequisite of good medicinal herbs. However, there exist numerous indices for quality evaluation and control in Chinese medicinal materials. However, most of these indices are non-interrelated each other, as well as having little relationship with efficiency and safety. The results of different evaluatior methods may not be consistent, even contradictory. Considering the complex material properties of Chinese medicinal materials, single method and index is difficult to objectively and comprehensively reflect the quality. Therefore, it is essential to explore the integrated evaluation methods. In this paper, oriented by the integrated evaluation strategies for traditional Chinese medicine quality, a new method called integrated quality index (IQI) by the integration of empirical evaluation, chemical evaluation, and biological evaluation was proposed. In addition, a study case of hypertoxic herb Aconitum carmichaelii Debx. was provided to explain this method in detail. The results suggested that in the view of specifications, the average weight of Jiangyou aconite was the greatest, followed by Weishan aconite, Butuo aconite, Hanzhong aconite, and Anxian aconite; from the point of chemical components, Jiangyou aconite had the characteristic with strong efficacy and weak toxicity, next was Hanzhong aconite, Butuo aconite, Weishan aconite, and Anxian aconite; taking toxicity price as the index, Hanzhong aconite and Jiangyou aconite have the lower toxicity, while Butuo aconite, Weishan aconite, and Anxian aconite have the relatively higher one. After the normalization and integration of evaluation results, we calculated the IQI value of Jiangyou aconite, Hanzhong aconite, Butuo aconite, Weishan aconite, and Anxian aconite were 0.842 +/- 0.091, 0.597 +/- 0.047, 0.442 +/- 0.033, 0.454 +/- 0.038, 0.170 +/- 0.021, respectively. The quality of Jiangyou aconite is significantly better than the others (P < 0.05) followed by Hanzhong aconite, which is consistent with the traditional understanding of genuineness. It can be concluded that IQI achieves the integrated control and evaluation for the quality of Chinese medicinal materials, and it is an exploration for building the good medicinal herbs standards. In addition, IQI provides technical supports for the geoherbalism evaluation, selective breeding, the development of precision decoction pieces, high quality and favourable price in market circulation, and rational drug use.
Aconitum ; chemistry ; Drugs, Chinese Herbal ; standards ; Medicine, Chinese Traditional ; standards

To study the effects of ouabain on the inner ear glial cells, and to lay the foundation for the study of stem cell transplantation in the treatment of sensorineural hearing loss.Methods Sixty adult female SPF grade CBA / J mice were randomly divided into experimental group and control group, with 30 mice in each group.Animals in the experimental group received 3mM ouabain via the round window membrane, while mice in control group received normal saline.The mice were sacrificed at 7 days, 14 days and 30 days after the administration,respectively.Immunofluorescence histochemical staining was used to detect the inner ear glial cells in spiral ganglion.Results Some inner ear glial cells survived in the spiral ganglion of the experimental group, while with decreased numbers and disorganized structure compared to those of in the control group.Comparing to those of in the control group, the number and density of inner ear glial cells in the experimental group were significantly decreased from 7 days afterouabain administration,further decreased at 14 days and reduced to the lowest at 30 days after ouabain administration, the differences between the 2 groups were statistically significant (P < 0.05).Among the experimental group, the number of inner ear glial cells at 30 days was significantly decreased when compared to those of at 7 days and 14 days, respectively.Conclusion Application of ouabain to mouse inner ear via the round window membrane leads to an acute and progressive direct damage to the inner ear glial cells in the spiral ganglion.

Objective To explore the effect of complement activation on bone marrow mesenchymal stem cells (BMSCs)and evaluate the effect after transfection of complement regulatory proteins.Methods Bone marrow aspirate was harvested from 10 cases of patients suffered from fractures.Mesenchymal stem ceils were isolated,indentified cultured and then experimented in vitro.The complement cytotoxicity on the mesenchymal stem cells in autologous serum was measured by Europium cytotoxicity assay.The samples were divided into BMSCs group,BMSCs+ autologous human serum (AHS) group and BMSCs+ inactivated autologous human serum (iAHS) group.The complement membrane attack complex (MAC) deposited on the membranes was detected by flow cytometry.Finally,the cytotoxicity on BMSCs was measured after transfected with membrane complement regulatory proteins (mCRPs).All samples were divided into BMSCs with mCRPs untransfected group and BMSCs with mCRPs transfected group.Results More than 95％ of cells derived from bone marrow were identified to be mesenchymal stem cells through detection of cell surface markers by flow cytometry.The cytotoxicity of untreated cells was 0.41％± 1.48％.BMSCs harvested from the 10 patients all had cytotoxicity after incubated with autologous serum,and the cytotoxicity was 32.59％±2.73％,while cytotoxicity after incubated with complement inactivated autologous serum was 2.59％±3.08％,which was similar to control group.Complement attack complex (MAC) could be detected on the BMSCs incubated with autologous serum,which implied the complement activation.After transfection of mCRPs,the cytotoxicity of autologous serum on transfected cells was decreased.The cytotoxicity of untransfected cells (41.70％±4.47％) had significant difference compared to the cells transfected with CD55 (21.87％±2.19％),the cells transfected with CD59 (18.67％± 1.42％),and the cells transfected with CD46+CD55+CD59 (28.43％±2.14％).CD55,CD59 and CD46+CD55 +CD59 transfected groups could impair effectively the cytotoxicity from complement.However,the cytotoxicity impairment was less effective in CD46 transfected cells (39.30％±3.96％),which had no significant difference compared to untransfected cells.Conclusion Membrane complement regulatory proteins could effectively protect bone marrow mesenchymal stem cells from attacks by complement.

· AIM: To investigate the expression and the possible implication of CD40/CD40L costimulatory molecules in erythema nodosum of patients with Beh(c)et's disease.· METHODS: Sampling was done from erythema nodosum of 5 patients with Beh(c)et's disease and normal skin of 2 healthy individuals. Immunohistochemical staining was performed to examine the expression of CD4, CD8, CD19, CD68, HLA-DR,CD40 and CD40L molecules in the obtained tissues.· RESULTS: Approximately 90% of epidermic cells in erythema nodosum expressed CD40 molecule. In the dermis and subcutaneous tissue, a significantly increased number of CD4+Tcells, CD8+Tcells, CD19+cells, CD68+cells, HLA-DR+cells,CD40L+cells, and CD40+cells were observed in the erythema nodosum as compared with that in normal skin. Double staining showed that CD40L molecules were expressed on 45% of CD4+T cells. CD40 molecules were expressed on 100% CD68+ cells and 59.2% of HLA-DR+cells respectively.· CONCLUSION: A number of CD40/CD40L costimulatory molecules are upreguiated in the erythema nodosum of patients with Behcet's disease.

Objective To identify the species of sarcosaphagous flies by amplifying cytochrome oxidase subunitI (CO I) gene fragment,combined with morphological characteristics.Methods The DNA of sarcosaphagous flies was extacted by modified Tris balanced phenol-Tris saturated phenol protocol,the amplificationand sequencing of CO Ⅰ fragment were conducted,and the results were then compared with the database for analysis.Results The modified DNA extration method by Tris balanced phenol-Tris saturated phenol could obtain effective DNA of sarcosaphagous flies,and be applied for CO Ⅰ fragment amplification,and thus for identification of the species of sarcosaphagous flies.Conclusion The DNA extracted from sarcosaphagous flies by modified Tris balanced phenol-Tris saturated phenol protocol could be used astemplate for the amplification of CO Ⅰ fragment,and the system could identify the species of sareosaphagous flies after sequencing and alignment with the sequences of database.Compared with traditional identification methods of using morphological characteristics,the current system is more accurate and could be more widely applied.