Exploring the protective mechanism of Tibetan medicine Potentilla anserine on cyclophosphamide-induced myelosuppression based on metabonomics technology / 药学学报

The study aims to explore the effects and mechanisms of water extract of Potentilla anserina (PA) on myelosuppression mice induced by cyclophosphamide based on metabonomics. The myelosuppressive mouse model was established by injected with cyclophosphamide and treated with water extract of PA. Thymus and spleen indexes, peripheral hemogram and bone marrow nucleated cells of each group was detected. Bone marrow pathology analysis was performed by hematoxylin-eosin staining. The levels of interleukin 3 (IL-3), interleukin 6 (IL-6), erythropoietin (EPO), granulocyte colony stimulating factor (GM-CSF), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in serum were measured. The changes of biomarkers and related metabolic pathways were analyzed by UPLC-Q-TOF/MS-based metabonomics. Animal experiments were approved by the Animal Ethics Committee of Southwest Minzu University. The high doses of PA could significantly improve the decrease of white blood cell (WBC), red blood cell (RBC) counts and hemoglobin (HGB) levels of mice induced by cyclophosphamide (P < 0.05), and significantly increase the number of nucleated cells and the area of hematopoietic tissue in femoral bone marrow. The medium and high doses of PA could significantly improve the serum levels of SOD, CAT, MDA, IL-6 and GM-CSF (P < 0.05), and have no significant effect on the expression of IL-3 and EPO (P > 0.05). Serum metabolomics analysis showed that the aqueous extracts of PA could alleviate myrosuppression by regulating the aminoacyl-tRNA, valine, leucine and isoleucine biosynthesis mediated by 13 different metabolites such as valine, leucine, asparagine and hydroxyisohexic acid. PA improve the inhibition of hematopoietic function in myelosuppression mouse, and its mechanisms may be related to anti-oxidation and promoting the expression of hematopoietic-related cytokines and regulating the related metabolic pathways.

Mechanism of Tibetan medicine Ershiwuwei Shanhu Pills on scopolamine-induced learning and memory impairment in mice based on Keap1/Nrf2/HO-1 signaling pathway / 中国中药杂志

This study aims to investigate the mechanism of the Tibetan medicine Ershiwuwei Shanhu Pills(ESP) in improving scopolamine-induced learning and memory impairment in mice based on Keap1/Nrf2/HO-1 signaling pathway. ICR mice were randomized into blank group, model group, low-dose(200 mg·kg~(-1)), medium-dose(400 mg·kg~(-1)), and high-dose(800 mg·kg~(-1)) ESP groups, and donepezil hydrochloride group. The learning and memory impairment was induced in mice by intraperitoneal injection of scopola-mine. The learning and memory abilities of mice were detected by Morris water maze test, and the damage of hippocampal neurons and cortical neurons was detected based on Nissl staining. The expression of neuron specific nuclear protein(NeuN) in hippocampus and cortex of mice was determined by immunofluorescence assay, and the content of acetylcholine(Ach) and the activity of acetylcholines-terase(AchE) in hippocampus of mice by kits. Moreover, the content of superoxide dismutase(SOD), malondialdehyde(MDA), catalase(CAT), and total antioxidant capacity(T-AOC) in serum of mice was detected. The content of Kelch-like ECH-associated protein 1(Keap1), nuclear factor erythroid 2-related factor 2(Nrf2), and heme oxygenase 1(HO-1) in hippocampus was determined by Western blot. The results showed that there were significant differences in the trajectory map of mice among different groups in the behavioral experiment. Moreover, the latency of ESP groups decreased significantly compared with that in the model group. The hippocampal neurons in the high-dose ESP group were significantly more than those in the model group and the cortical neurons in the high-dose and medium-dose ESP groups were significantly more than those in the model group. The expression of NeuN in the model group was significantly decreased compared with that in the blank group, and the expression in the ESP groups was significantly higher than that in the model group. The AchE activity and MDA level were significantly decreased, and Ach content and levels of SOD, CAT, and T-AOC in the ESP groups were significantly increased in the ESP groups compared with those in the model group. The expression of Keap1 in the model group was significantly increased compared with that in the blank group, and the Keap1 expression increased insignificantly in ESP groups compared with that in the model group. The expression of Nrf2 and HO-1 was significantly lower in the model group than in the blank group, and the expression was significantly higher in the medium-dose ESP group than in the model group. In conclusion, ESP protected mice against the scopolamine-induced learning and memory impairment by regulating the Keap1/Nrf2/HO-1 signaling pathway.

Ershiwuwei Shanhu Pills regulate Akt/mTOR/GSK-3β signaling pathway to alleviate Alzheimer's disease mice / 中国中药杂志

The present study investigated the mechanism of the Tibetan patent medicine Ershiwuwei Shanhu Pills(ESP) in alleviating Alzheimer's disease in mice via Akt/mTOR/GSK-3β signaling pathway. BALB/c mice were randomly assigned into a blank control group, a model group, low(200 mg·kg~(-1)), medium(400 mg·kg~(-1)) and high(800 mg·kg~(-1)) dose groups of ESP, and donepezil hydrochloride group. Except the blank control group, the other groups were given 20 mg·kg~(-1) aluminum chloride by gavage and 120 mg·kg~(-1) D-galactose by intraperitoneal injection for 56 days to establish Alzheimer's disease model. Morris water maze was used to detect the learning and memory ability of mice. The level of p-tau protein in mouse hippocampus and the levels of superoxide dismutase(SOD), malondialdehyde(MDA), catalase(CAT), and total antioxidant capacity(T-AOC) in hippocampus and serum were detected. Hematoxylin-eosin staining and Nissl staining were performed for the pathological observation of whole brain in mice. TdT-mediated dUTP nick-end labeling(TUNEL) staining was employed for the observation of apoptosis in mouse cortex. Western blot was adopted to detect the protein levels of p-mTOR, p-Akt, and GSK-3β in the hippocampus. Compared with the model group, the ESP groups showcased alleviated pathological damage of the whole brain, decreased TUNEL positive cells, reduced level of p-tau protein in hippocampus, and risen SOD, CAT, and T-AOC levels and declined MDA level in hippocampus and serum. Furthermore, the ESP groups had up-regulated protein levels of p-mTOR and p-Akt while down-regulated protein level of GSK-3β in hippocampus. Therefore, ESP can alleviate the learning and memory decline and oxidative damage in mice with Alzheimer's disease induced by D-galactose combined with aluminum chloride, which may be related to Akt/mTOR/GSK-3β signaling pathway.

Mechanism of Tibetan medicine Ershiwuwei Songshi Pills against liver injury induced by acetaminophen in mice based on Keap1/Nrf2 and TLR4/NF-κB p65 signaling pathways / 中国中药杂志

The present study investigated the mechanism of the Tibetan medicine Ershiwuwei Songshi Pills(ESP) against the liver injury induced by acetaminophen(APAP) in mice based on the kelch-like ECH-associated protein 1(Keap1)/nuclear transcription factor E2 related factor 2(Nrf2) and Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB) p65 signaling pathways. Kunming mice were randomly divided into a blank control group, a model group, an N-acetyl-L-cysteine(NAC) group, and high-(400 mg·kg~(-1)), medium-(200 mg·kg~(-1)), and low-dose(100 mg·kg~(-1)) ESP groups. After 14 days of continuous administration, except for those in the control group, the mice were intraperitoneally injected with 200 mg·kg~(-1) APAP. After 12 h, the serum and liver tissues of mice were collected. Hematoxylin-eosin(HE) staining was performed on pathological sections of the liver, and the levels of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in the serum and the levels of glutathione(GSH), malondialdehyde(MDA), superoxide dismutase(SOD), catalase(CAT), myeloperoxidase(MPO), and total antioxidant capacity(T-AOC) in liver tissue homogenate were detected to observe and analyze the protective effect of ESP on APAP-induced liver injury in mice. The serum levels of tumor necrosis factor-alpha(TNF-α), interleukin-1 beta(IL-1β), and interleukin-6(IL-6) were determined by enzyme-linked immunosorbent assay(ELISA). The protein expression of Nrf2, Keap1, TLR4, and NF-κB p65 in the liver was determined by Western blot. Quantitative real-time was used to determine the mRNA expression of glutamate-cysteine ligase catalytic subunit(GCLC), glutamate-cysteine ligase regulatory subunit(GCLM), heme oxygenase-1(HO-1), and NAD(P)H dehydrogenase quinone 1(NQO-1) in the liver to explore the mechanism of ESP in improving APAP-induced liver damage in mice. As revealed by results, compared with the model group, the ESP groups showed improved liver pathological damage, decreased ALT and AST levels in the serum and MDA and MPO content in the liver, increased GSH, SOD, CAT, and T-AOC in the liver, reduced TNF-α and IL-6 levels in the serum, down-regulated expression of Keap1 in the liver cytoplasm and NF-κB p65 in the liver nucleus, up-regulated expression of Nrf2 in the liver nucleus, insignificant change in TLR4 expression, and elevated relative mRNA expression levels of antioxidant genes GCLC, GCLM, HO-1, and NQO-1. ESP can reduce the oxidative damage and inflammation caused by APAP, and the mechanism may be related to the Keap1/Nrf2 signaling pathway and the signal transduction factors on the TLR4/NF-κB p65 pathway.

Research progress on precious Tibetan medicine formula in prevention and treatment of central nervous system diseases / 中国中药杂志

Precious Tibetan medicine formula is a characteristic type of medicine commonly used in the clinical treatment of central nervous system diseases. Through the summary of modern research on the precious Tibetan medicine formulas such as Ratnasampil, Ershiwuwei Zhenzhu Pills, Ershiwewei Shanhu Pills, and Ruyi Zhenbao Pills, it is found that they have obvious advantages in the treatment of stroke, Alzheimer's disease, epilepsy, angioneurotic headache, and vascular dementia. Modern pharmacological studies have shown that the mechanisms of precious Tibetan medicine formulas in improving central nervous system diseases are that they promote microcirculation of brain tissue, regulate the permeability of the blood-brain barrier, alleviate inflammation, relieve oxidative stress damage, and inhibit nerve cell apoptosis. This review summarizes the clinical and pharmacological studies on precious Tibetan medicine formulas in prevention and treatment of central nervous system diseases, aiming to provide a reference for future in-depth research and innovative discovery of Tibetan medicine against central nervous diseases.

A preliminary cervical cancer screening cascade for eight provinces rural Chinese women: a descriptive analysis of cervical cancer screening cases in a 3-stage framework / 中华医学杂志(英文版)

Background@#Cascade analysis is an effective method to analyze the processing data of an event, such as a provided service or a series of examinations. This study aimed to develop a primary cervical cancer screening cascade in China to promote the quality of the screening process.@*Methods@#We designed a cervical cancer screening cascade in China according to the program flow chart. It had three stages, each with two steps and one result. Data from 117,522 women aged 35 to 64 years in the Rural Cervical Cancer Surveillance Project from January 1, 2014, to December 31, 2014, were collected to analyze the main results of the cascade. The data and proportion are used to describe the follow-up of cervical cancer and pre-cancer detection rate.@*Results@#In 2014, 117,522 (80.94% of all cases reported by the Rural Cervical Cancer Surveillance Project) women aged 35 to 64 years had not received cervical cytology in the previous 3 years. The pre-cancer and cancer detection rates were 256.12/100,000 and 16.16/100,000, respectively. A total of 3031 cases failed to follow-up through the screening process, and 1189, 1555, and 287 cases were lost at cervical cytology, colposcopy, and histopathological screening stages, respectively. The estimated cases of pre-cancer and cancer cases would have been 544 and 34, respectively, and the estimated detection rates of pre-cancer and cancer would have been 462.89/100,000 and 28.93/100,000, respectively.@*Conclusion@#In order to increase the detection rate of cervical cancer, cervical cancer screening staff should focus on increasing the rate of follow-up of those who are positive for cervical cancer screening (ie, those with positive cytology results), especially for the 40 to 44 years age range.

A preliminary cervical cancer screening cascade for eight provinces rural Chinese women: a descriptive analysis of cervical cancer screening cases in a 3-stage framework / 中华医学杂志(英文版)

BACKGROUND@#Cascade analysis is an effective method to analyze the processing data of an event, such as a provided service or a series of examinations. This study aimed to develop a primary cervical cancer screening cascade in China to promote the quality of the screening process.@*METHODS@#We designed a cervical cancer screening cascade in China according to the program flow chart. It had three stages, each with two steps and one result. Data from 117,522 women aged 35 to 64 years in the Rural Cervical Cancer Surveillance Project from January 1, 2014, to December 31, 2014, were collected to analyze the main results of the cascade. The data and proportion are used to describe the follow-up of cervical cancer and pre-cancer detection rate.@*RESULTS@#In 2014, 117,522 (80.94% of all cases reported by the Rural Cervical Cancer Surveillance Project) women aged 35 to 64 years had not received cervical cytology in the previous 3 years. The pre-cancer and cancer detection rates were 256.12/100,000 and 16.16/100,000, respectively. A total of 3031 cases failed to follow-up through the screening process, and 1189, 1555, and 287 cases were lost at cervical cytology, colposcopy, and histopathological screening stages, respectively. The estimated cases of pre-cancer and cancer cases would have been 544 and 34, respectively, and the estimated detection rates of pre-cancer and cancer would have been 462.89/100,000 and 28.93/100,000, respectively.@*CONCLUSION@#In order to increase the detection rate of cervical cancer, cervical cancer screening staff should focus on increasing the rate of follow-up of those who are positive for cervical cancer screening (ie, those with positive cytology results), especially for the 40 to 44 years age range.

Characteristics of patients with fluid extravasation during retrograde ureteroscopic holmium laser lithotripsy for renal calculi

To explore the risk factors of fluid extravasation during retrograde ureteroscopic holmium laser lithotripsy for renal calculi. Three hundred and twenty-seven patients with renal calculi ranging 10 to 20 mm received retrograde ureteroscopic holmium laser lithotripsy at Renmin Hospital of Wuhan University, Wuhan, China from January 2004 to December 2010. The clinical records were reviewed, and the correlation was studied between various clinical factors and fluid extravasation complications during operation. The clinical factors to be tested included patients' gender and age [<30, 30-50, and >50 years], hydronephrosis degree, previous intervention for renal calculi [none, shock-wave lithotripsy, and open surgery], upper urinary tract infection, ureteral access sheath placement, and procedure duration [<60, 60-120, and >120 mins]. The data were processed by SPSS Version 16.0 statistical software, x[2] test, and binary logistic regression were used for analysis. Fluid extravasation complications appeared in 35 patients. Patients` gender, age, and hydronephrosis degree were irrelevant to the occurrence of fluid extravasation, while having previous open surgery for renal calculi, without ureteral access sheath placement, upper urinary tract infection, and long procedure duration were all responsible for higher incidence of the complications. Reasonable selection of patients, effective control of upper urinary tract infection, routine ureteral access sheath placement, and controlling procedure duration help to decrease the incidence of fluid extravasation complications in retrograde ureteroscopic holmium laser lithotripsy for renal calculi

Murine model of busulfan-induced spermatogenesis regeneration: a quantitative evaluation / 中华男科学杂志

<p><b>OBJECTIVE</b>To quantitatively evaluate the murine model of spermatogenesis regeneration induced by two-dose busulfan injection.</p><p><b>METHODS</b>Fifty-four male mice were randomly divided into a control and two model groups of equal number, the former treated by two-dose intraperitoneal injection of 50% DMSO solution at 10 ml/kg, and the latter by that of busulfan at 10 mg/kg and 15 mg/kg respectively to establish spermatogenesis regeneration models, both at the interval of 24 days between the two doses. Spermatogenesis in seminiferous epithelia was evaluated by Johnsen score, and the expressions of GATA-4 and GDNF mRNA in Sertoli cells were detected by real time quantitative PCR at 3, 4 and 8 weeks after the treatment.</p><p><b>RESULTS</b>Johnsen score kept stable in the control group at all stages (P > 0.05), but higher than in the model groups at 3 and 4 weeks (P < 0.01). It was lower in the 15 mg/kg than in the 10 mg/kg model group at 4 and 8 weeks (P < 0.01) , and than in the control group at 8 weeks (P < 0.05), but had no significant difference between the 10 mg/kg and the control groups (P > 0.05). Nor did the expression of GATA-4 mRNA in Sertoli cells show any significant difference among the three groups at different stages after the treatment (P > 0.05), and that of GDNF mRNA at different stages in the control group (P > 0.05). Compared with the controls, the level of GDNF mRNA in Sertoli cells was significantly higher at 3 weeks but lower at 4 weeks in the model groups (P < 0.01), and lower in the 15 mg/kg group (P < 0.01) and comparable in the 10 mg/kg group at 8 weeks (P > 0.05); and it was lower in the 15 mg/kg than in the 10 mg/kg group at all stages (P < 0.01).</p><p><b>CONCLUSION</b>Two-dose intraperitoneal injection of 10 mg/kg busulfan at the interval of 24 days is an optimal option for the establishment of a murine model of spermatogenesis regeneration. Higher dose of busulfan may induce deficient expression of GDNF in Sertoli cells and result in incomplete restoration of spermatogenesis.</p>

Comparison of baicalin,paeoniflorin and glycyrrhetic acid in different decoctions of Huangqin Decoction / 中成药

AIM: To compare the contents of baicalin,paeoniflorin and glycyrrhetic acid in the seperated and mixed decoction of Huangqin Decoction(Radix Scutellariae,Radix Paeoniae alba,Radix et Rhizoma Glycyrrhizae and Fructus Jujubae). METHODS: The contents of baicalin,paeoniflorin and glycyrrhetic acid were analyzed by HPLC.Chromatographic conditions included: Hypersil BDS C_(18) column and the mobile phase consisted of a mixture of methanol-water-phosphoric acid(47(∶)53(∶)0.2),acetonitrile-water-phosphoric acid(18(∶)82(∶)0.1) and methanol-water-phosphate buffer solution(70(∶)29(∶)1).Baicalin,paeoniflorin and glycyrrhetic acid were detected at 280 nm,230 nm and 250 nm respectively. RESULTS: The linear range of baicalin,paeoniflorin and glycyrrhetic acid were 2.88—72.00 ?g/mL,2.85—22.80 ?g/mL and 4.16—52.00 ?g/mL respectively.The contents of baicalin in the mixed decoction was higher than that in the seperated decoction.The contents of paeoniflorin and glycyrrhetic acid in the mixed decoction were almost as same as that in seperated decoction. CONCLUSION: The contents of active ingredients in the mixed and seperated decoction are different.