ObjectiveBased on multi-index and multi-method detection， the quality of Rukuaixiao preparations was systematically evaluated from the perspective of characteristic components， and the existing problems were analyzed. MethodsLiquid chromatography-mass spectrometry（LC-MS） for the determination of 16 characteristic components was established to evaluate the quality of 129 batches of Rukuaixiao preparations. High performance liquid chromatography（HPLC） was established to determine the contents of salvianolic acids and tanshinones， investigate the rationality of quality control index of Salviae Miltiorrhizae Radix et Rhizoma in the standard for Rukuaixiao preparations in the 2020 edition of Pharmacopoeia of the People's Republic of China（hereinafter referred to as Chinese Pharmacopoeia） （volume Ⅰ）， and trace the causes of significant difference among different batches. The processing and different extraction methods of Vaccariae Semen were tested， analyzing the impact of formulation changes across different editions of Chinese Pharmacopoeia. The LC-MS was established for determining the changes in the ratio of toosendanin and isotoosendanin after water extraction of Toosendan Fructus. The contents of active components in different parts of Gleditsiae Spina were determined to identify the reason of the low contents of characteristic components in some enterprises. ResultsBased on the comprehensive analysis of the samples from different dosage forms， the contents of characteristic components of Vaccariae Semen and Gleditsiae Spina in tablets from manufacturer B and granules from manufacturer D were significantly higher than those in tablets from manufacturer A， and tablets and capsules from manufacturer C. The contents of tanshinones in some batches of products from manufacturer A were abnormally high， potentially linked to the use of 70% ethanol reflux during extraction of Salviae Miltiorrhizae Radix et Rhizoma. All samples from manufacturer C exhibited abnormally high proportions of salvianic acid A and 3，4-dihydroxybenzaldehyde（salvianolic acid degradation products） to the total seven phenolic acids， indicating a risk of blindly pursuing tanshinol content and neglecting more effective components. The extraction rate of the characteristic components from Vaccariae Semen by decocting with water was obviously higher than that by reflux extraction with 70% ethanol. However， using the stir-fried Vaccariae Semen could reduce the loss of ingredients. The content ratio of toosendanin and isotoosendanin decreased from the crude herb to the prepared medicine when Toosendan Fructus was prepared by water decoction. The reason for the low component content of Gleditsiae Spina may be attributed to manufacturers using excessive non-medicinal parts in their formulations. ConclusionIt is suggested that enterprises should ensure the quality of raw material inputs， especially those without quality-control items in the standard， reduce the use of non-medicinal parts， and prohibit arbitrary alterations to manufacturing methods or processes. It is also recommended that Vaccariae Semen in Rukuaixiao capsules and granules should be changed to the stir-fried processed products. Isotoosendanin should be taken into consideration in the drug supervision when Toosendan Fructus is prepared into medicine by water decoction. Salvianolic acid B should be set as a quality control index for Salviae Miltiorrhizae Radix et Rhizoma when revising the pharmacopoeia standard of Rukuaixiao preparations.

ObjectiveBased on multi-index and multi-method detection， the quality of Rukuaixiao preparations was systematically evaluated from the perspective of characteristic components， and the existing problems were analyzed. MethodsLiquid chromatography-mass spectrometry（LC-MS） for the determination of 16 characteristic components was established to evaluate the quality of 129 batches of Rukuaixiao preparations. High performance liquid chromatography（HPLC） was established to determine the contents of salvianolic acids and tanshinones， investigate the rationality of quality control index of Salviae Miltiorrhizae Radix et Rhizoma in the standard for Rukuaixiao preparations in the 2020 edition of Pharmacopoeia of the People's Republic of China（hereinafter referred to as Chinese Pharmacopoeia） （volume Ⅰ）， and trace the causes of significant difference among different batches. The processing and different extraction methods of Vaccariae Semen were tested， analyzing the impact of formulation changes across different editions of Chinese Pharmacopoeia. The LC-MS was established for determining the changes in the ratio of toosendanin and isotoosendanin after water extraction of Toosendan Fructus. The contents of active components in different parts of Gleditsiae Spina were determined to identify the reason of the low contents of characteristic components in some enterprises. ResultsBased on the comprehensive analysis of the samples from different dosage forms， the contents of characteristic components of Vaccariae Semen and Gleditsiae Spina in tablets from manufacturer B and granules from manufacturer D were significantly higher than those in tablets from manufacturer A， and tablets and capsules from manufacturer C. The contents of tanshinones in some batches of products from manufacturer A were abnormally high， potentially linked to the use of 70% ethanol reflux during extraction of Salviae Miltiorrhizae Radix et Rhizoma. All samples from manufacturer C exhibited abnormally high proportions of salvianic acid A and 3，4-dihydroxybenzaldehyde（salvianolic acid degradation products） to the total seven phenolic acids， indicating a risk of blindly pursuing tanshinol content and neglecting more effective components. The extraction rate of the characteristic components from Vaccariae Semen by decocting with water was obviously higher than that by reflux extraction with 70% ethanol. However， using the stir-fried Vaccariae Semen could reduce the loss of ingredients. The content ratio of toosendanin and isotoosendanin decreased from the crude herb to the prepared medicine when Toosendan Fructus was prepared by water decoction. The reason for the low component content of Gleditsiae Spina may be attributed to manufacturers using excessive non-medicinal parts in their formulations. ConclusionIt is suggested that enterprises should ensure the quality of raw material inputs， especially those without quality-control items in the standard， reduce the use of non-medicinal parts， and prohibit arbitrary alterations to manufacturing methods or processes. It is also recommended that Vaccariae Semen in Rukuaixiao capsules and granules should be changed to the stir-fried processed products. Isotoosendanin should be taken into consideration in the drug supervision when Toosendan Fructus is prepared into medicine by water decoction. Salvianolic acid B should be set as a quality control index for Salviae Miltiorrhizae Radix et Rhizoma when revising the pharmacopoeia standard of Rukuaixiao preparations.

AIM: To explore the roles of heat shock protein 90 (HSP90) in the blockage of hydrogen sulfide (H2S) against chemical hypoxia-mimetic agent (cobalt chloride, CoCl_2)-induced oxidative stress injuries in H9c2 cardiac cell. METHODS: H9c2 cells were treated with CoCl_2 to set up the chemical hypoxia-induced the model of cardiomyocyte injury. Sodium hydrosulfide (NaHS, a H2S donor) was added into medium for 30 min before CoCl_2 treatment. ATP content was detected by high performance liquid chromatogram (HPLC). Mitochondrial membrane potential (MMP) was measured by rhodamine123 (Rh123) staining and photofluorography. The activity of superoxide dismutase (SOD) was observed using a SOD kit. The expression of heme oxygenase-1 (HO-1) was evaluated by Western blotting. RESULTS: CoCl_2 at concentration of 600 μmol/L significantly reduced SOD activity, ATP level and MMP, and enhanced the expression of HO-1 in H9c2 cells. Pretreatment with 400 μmol/L NaHS dramatically inhibited the cytotoxicity induced by CoCl_2, increased SOD activity, ATP level and MMP, decreased HO-1 expression. 17-allylamino-17 demethoxygeldanamycine(17AAG), an inhibitor of HSP90, obviously blocked the inhibitory effect of H2S on the CoCl_2-induced cytotoxicity, reduced the levels of ATP and MMP, increased HO-1 expression. However, no significantly influence on SOD activity was observed. CONCLUSION: HSP90 may mediate the cardioprotection of H2S via inhibiting the oxidative stress induced by chemical hypoxia.

Objective: To investigate the effect of vitamin B12 on the growth and hemopoiesis of broilers. Methods: The Arbor Acres broilers were fed the experimental diets with different doses of vitamin B12 0.00, 0.008, 0.016, 0.024 mg/kg for 3 weeks. The body weight gain and hematologic indices of the broilers were measured. Results: Vitamin B12 deficiency in the diet could depress the body weight gain and feed conversion ratio of the broilers, decrease the amount of red blood cell and hemoglobin, enlarge the volume of red blood cell, increase the content of each cell hemoglobin, reduce platelet in blood. While 0.008 mg/kg vitamin B12 was added in the experimental diets, the body weight gain and feed conversion ratio of the broilers increased markedly. Furthermore, when 0.016mg/kg vitamin B12 was added to the diet, the number of the red blood cell and the content of hemoglobin of the broilers increased significantly, and the shape and volume of the red blood cell became ordinary. There were significants interrelations between BWG,RBC,Hb,PCV,PLT,MCV,MCH of the broilers and the addition of vitamin B12 in the diet. Conclusion: Vitamin B12 deficiency could result in the megaloblastic animia. 0.02-0.03 mg vitamin B12 per kilo diet was required to maintain the growth and normal hemopoiesis of the broilers.