Zuogui pills maintain the stemness of oogonial stem cells and alleviate cyclophosphamide-induced ovarian aging through Notch signaling pathway.

BACKGROUND: Zuogui pills (ZGP), a classical prescription of traditional Chinese medicine, have been widely used in the treatment of ovarian aging. Previous studies have demonstrated its efficacy on protecting ovarian aging, and the mechanisms were mostly relevant to inhibiting the apoptosis of follicles and activating the primordial follicles. However, whether ZGP could stimulate the oogonial stem cells (OSCs) to refresh the follicle pool remains poorly understood. PURPOSE: To investigate the effects of ZGP on the stemness of OSCs in cyclophosphamide (Cy)-induced ovarian aging. STUDY DESIGN AND METHODS: Female Sprague-Dawley (SD) rats were randomly divided into 8 groups: control group, model group, ZGP groups (low / high dose groups), estradiol valerate (EV) groups (low / high dose groups), DAPT group and DAPT+ZGP-L group. After modeling with Cy, the ZGP groups and EV groups were treated with ZGP and EV for 8 weeks respectively. Meanwhile, the DAPT groups were treated with DAPT twice a week. Additionally, OSCs were also isolated after modeling, and then treated with drug serum containing ZGP or EV. Ovarian volume and the ratio of weight of total ovaries to the body weight were measured. The serum hormones were measured by ELISA. Quantities and location of OSCs in ovaries were detected by flow cytometry and immunofluorescence. Cell viability was measured by CCK8. And OSCs were identified by immunofluorescence. Biomarkers of germ cells, stem cells and associated to differentiation and meiosis were detected by qPCR and western blot. Proteins in Notch signaling pathway were detected by western blot and immunofluorescence. RESULTS: After treating with ZGP, ovarian volume and the ratio of weight of total ovaries to the body weight increased. ZGP could increase serum AMH and E2 level and decrease serum FSH level. Quantities and cell viability of OSCs increased after ZGP treatment in vivo and in vitro. In addition, treatment with ZGP could increase not only the expression of MVH, Oct4 and DAZL, but also the expression of ZP1 and ZP2. Furthermore, ZGP could up-regulate the expression of Notch intracellular domain (NICD), HES1 and HES5. After blocking the Notch signaling pathway, ZGP could increase not only the expression of NICD, HES1 and HES5, but also the expression of MVH, Oct4, DAZL, ZP1 and ZP3. CONCLUSION: In conclusion, the mechanism of ZGP on treating ovarian aging may be relevant to maintain the stemness of OSCs by up-regulating Notch signaling pathway, which added the mechanism of ZGP on the perspective of OSCs at first time.

Guizhi Gancao Longgu Muli decoction for insomnia: A protocol for a systematic review.

BACKGROUND: Insomnia is a prevalent and bothersome disorder of sleep initiation and maintenance. Although efficacious treatments for insomnia have been available for decades, they all have their own limitations. Guizhi Gancao Longgu Muli Decoction (GGLMD), a popular complementary and alternative therapy, has been widely applied to treat insomnia in some Asian countries for centuries. Yet no systematic reviews have comprehensively assessed the efficacy and safety of GGLMD as a treatment for insomnia. METHODS: A comprehensive search up to November, 2019 will be conducted in the following electronic databases: the Cochrane Library, Embase, PubMed, Web of Science, the Chinese National Knowledge Infrastructure (CNKI), the Chinese Biomedical Literature Database (CBM), the Chinese Scientific Journal Database (VIP), and the Wanfang Database. The primary outcomes will be sleep quality including Pittsburgh Sleep Quality Index (PSQI) and polysomnography (PSG). Stata 15 will be used for data analysis as well. RESULTS: This study will provide the current evidence of insomnia treated with GGLMD from the several points including PSQI and PSG. CONCLUSION: The consequence of this summary will furnish proof to evaluate if GGLMD is effective in the treatment of insomnia. ETHICS AND DISSEMINATION: Without personal information involved, ethical approval and informed consent form is no need. The review will be submitted to a peer-reviewed journal prospectively to spread our findings. PROSPERO REGISTRATION NUMBER: PROSPERO CRD42018118336.

Transcriptomic profiles of 33 opium poppy samples in different tissues, growth phases, and cultivars.

Opium poppy is one of the most important medicinal plants and remains the only commercial resource of morphinan-based painkillers. However, little is known about the regulatory mechanisms involved in benzylisoquinoline alkaloids (BIAs) biosynthesis in opium poppy. Herein, the full-length transcriptome dataset of opium poppy was constructed for the first time in accompanied with the 33 samples of Illumina transcriptome data from different tissues, growth phases and cultivars. The long-read sequencing produced 902,140 raw reads with 55,114 high-quality transcripts, and short-read sequencing produced 1,923,679,864 clean reads with an average Q30 rate of 93%. The high-quality transcripts were subsequently quantified using the short reads, and the expression of each unigene among different samples was calculated as reads per kilobase per million mapped reads (RPKM). These data provide a foundation for opium poppy transcriptomic analysis, which may aid in capturing splice variants and some non-coding RNAs involved in the regulation of BIAs biosynthesis. It can also be used for genome assembly and annotation which will favor in new transcript identification.

[Research rules of famous veteran traditional Chinese medicine practitioners in treating infertility].

To explore the medication rules of famous veteran traditional Chinese medicine practitioners in treating infertility based on medical cases of infertility collected from book series of Hundred Traditional Chinese Medicine Clinicians of Hundred Years in China and Prescription Proven by Traditional Chinese Medicine Masters. Researchers extracted the information of prescriptions from these cases according to the inclusion and exclusion criteria. Then, Excel 2010, SPSS Clementine(ver.12.0) and SPSS(ver. 22.0) were adopted respectively for frequency analysis, association rules analysis, cluster analysis and factor analysis. Cluster analysis was carried out by Ochiai algorithm of binary variable data, which was a systematic clustering method. And principal component analysis was used for factor analysis. Besides, KMO test and Bartlett spherical test were used for factor adaptation test. Finally, 151 medical cases and 396 prescriptions were included in total. A total of 60 kinds of frequently used herbs were identified according to the results of frequency analysis for medication, they were mainly used for activating blood and resolving stasis, tonifying and clearing heat respectively. The association rules analysis found out 25 drug pair association rules and 14 3-drug combination association rules. A total of 15 medicine groups were extracted by cluster analysis. KMO test and Bartlett spherical test indicated that the method was suitable for factor analysis, and 21 common factors were respectively extracted by factor analysis. Association rules indicated the characteristics of the therapeutic methods, like tonifying Qi and replenishing blood. The famous veteran traditional Chinese medicine practitioners utilized modified Siwu Decoction for tonifying blood and preferred Atractylodis Macrocephalae Rhizoma(Baizhu) for tonifying Qi. The results of both cluster analysis and factor analysis demonstrated the characteristics of the therapies for tonifying kidney, activating blood, tonifying spleen and dispelling dampness. In addition, factor analysis could reflect the therapies for nourishing Yin, tonifying kidney, warming the meridian, dissipating cold, nourishing blood and dispelling blood stasis. These results of analysis comprehensively showed out the medication characteristics of famous veteran traditional Chinese medicine practitioners of strictly following the pathogenesis, making good use of classical formulas and providing proper compatibility. In conclusion, data mining techniques(including frequency analysis, association rules analysis, cluster analysis and factor analysis) were used to comprehensively analyze the medication rules of famous veteran traditional Chinese medicine practitioners in treating infertility, which is helpful for guiding the clinical practice of treating infertility with traditional Chinese medicine.

[Famous veteran traditional Chinese medicine doctors' medication rules in treating spermatorrhea].

To explore famous veteran traditional Chinese medicine( TCM) doctors' medication rules in treating spermatorrhea based on the medical cases about spermatorrhea collected from Hundred TCM Clinical Doctors of Hundred Years in China and Proven Cases and National Medical Master Test Case. Researchers extracted such data as medicinal and therapies from these cases according to the inclusion and exclusion criteria. Then,Excle 2010,SPSS Clementine 12. 0 and SPSS 22. 0 were adopted respectively for frequency analysis,association rules analysis,cluster analysis and factor analysis. Cluster analysis was carried out by Ochiai algorithm of binary variable data,one of the systematic clustering methods. And principal component analysis was used for factor analysis. Finally,27 medical cases and 41 prescriptions were included in total. The frequency analysis showed that the most common therapy was the astringent therapy( n = 28),which was followed by the tonifying method,tranquilizing method,heat-clearing method,damp-dispelling method and Qiregulating method. The top three kinds of high-frequency traditional Chinese medicines were Poria,Ostreae Concha and Dioscoreae Rhizoma. The association rules analysis found out 11 association rules of medicine pairs,23 association rules of medicine combinations of the three kinds of herbs and 6 association rules of medicine combinations of five kinds of herbs. Among them,the representatives were Rubi Fructus→Fossilia Ossis Mastodi,Rehmanniae Radix Praeparata + Corni Fructus→Dioscoreae Rhizoma,and Pinelliae Rhizoma→Citri Reticulatae Pericarpium,which showed the therapeutic characteristics of astringing,tonifying the kidney and replenishing essence,fortifying the spleen and dispelling dampness. Moreover,7 medicine groups with relevance were extracted from the tree map generated by cluster analysis,including " Phellodendri Chinensis Cortex and Anemarrhenae Rhizoma" and " Angelicae Sinensis Radix,Rehmanniae Radix,Bupleuri Radix and Glycyrrhizae Radix et Rhizoma",which demonstrated the effects of nourishing Yin and purging fire,tonifying and soothing the liver. Besides,totally 12 common factors were obtained according to factor analysis,including 14 drug combinations,like " Amomi Fructus,Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex" and " Nelumbinis Stamen,Moutan Cortex,Poria and Scrophulariae Radix",which indicated the compatibility characteristics of tonifying the spleen and purging fire,dispelling dampness and clearing heat. In conclusion,data mining techniques( including frequency analysis,association rules analysis,cluster analysis and factor analysis) were used to comprehensively analyze the famous veteran TCM doctors' medication rules in treating spermatorrhea,which was helpful for guiding the clinical practice of treating spermatorrhea in TCM.

Efficacy and safety of grain moxibustion in hemiplegia: A systematic review and meta-analysis protocol.

BACKGROUND: The increasing morbidity of stroke brings enormous social and economic pressure to the countries. Hemiplegia is the common sequela of stroke patients, which affects their physical activities and daily life, and its optimal treatment is still an urgent problem. In Asian countries, moxibustion therapy is widely combined with rehabilitation in poststroke hemiplegia. Among them, grain moxibustion, due to its concentration on stimulating the acupoints and deep heat penetration, can promote neurorehabilitation after stroke. Therefore, based on the current literatures, the effectiveness and safety of grain moxibustion were systematically evaluated to provide possible alternative therapy for the rehabilitation of hemiplegia. METHODS: Studies search for eligible randomized controlled trials (RCTs) that use grain moxibustion as the sole treatment for hemiplegia and their data extraction will be done by two researchers. Mean difference (MD) or relative risk (RR) with fixed or random effect model in terms of 95% confidence interval (CI) will be adopted for the data synthesis. To evaluate the risk of bias, the Cochrane's risk of bias assessment tool will be utilized. The sensitivity or subgroup analysis will also be conducted when meeting high heterogeneity (I > 50%). RESULTS: This meta-analysis will provide an authentic synthesis of the grain moxibustion's effect for hemiplegia. CONCLUSION: The findings of the review offer updated evidence and identify whether grain moxibustion can be an effective treatment for hemiplegia. PROSPERO REGISTRATION NUMBER: PROSPERO CRD 42018117765.

Efficacy and safety of Zuogui Pill in treating osteoporosis: Study protocol of a systematic review.

BACKGROUND: Osteoporosis (OP) is a common skeletal disorder worldwide, resulting in increased bone fragility and high risk of fractures. The Zuogui Pill (ZGP), a classic Chinese herbal formulation, has played a vital role in the clinical practice of OP in China for centuries. Increasing studies have been performed for clarifying its anti-osteoporotic mechanisms. However, this treatment still lacks a systematic review for its efficacy and safety in the treatment of OP. METHODS: Eight electronic databases will be searched from inception to November 2018 by 2 independent researchers, in order to collect qualified randomized controlled trials (RCTs) on the ZGP treatment for OP. The therapeutic effects according to bone mineral density (BMD) will be adopted as the primary outcomes. RevMan V.5.3 software will be used for the data synthesis and the Cochrane's risk of bias assessment tool will be used to assess the risk of bias. RESULTS: This review will conduct a high-quality synthesis on present evidence of ZGP for OP. CONCLUSION: The conclusion of the study will indicate whether ZGP is an effective treatment for OP by providing updated evidence.PROSPERO registration number: PROSPERO CRD 42018114366.

Efficacy and safety of Xiaofeng powder (xiao feng san) in treating urticaria: A protocol for a systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Urticaria is a common skin disease that has a high impact on a patient's daily life. Xiaofeng powder (XFP) is one of the most commonly used Chinese herbal formula in China for urticaria. However, due to the lack of systematic evaluations, its clinical efficacy remains controversial. This meta-analysis was performed to evaluate the effect and safety of XFP for urticaria. METHODS: Seven databases, including Cochrane Central Register of Controlled Trials, PubMed, Embase, the Chinese National Knowledge Infrastructure (CNKI), the Chinese Scientific Journal Database (VIP), the Chinese Biomedical Literature Database (CBM), and the Wanfang Database. The period will be from their inception to September 2018. Randomized controlled trials of XFS used separately against conventional Western medicine therapy in patients with urticarial were included. After the methodologic quality was assessed and the valid data were extracted, RevMan 5.3 software was used for the final meta-analysis. RESULTS: The results will provide evidence regarding the efficacy and safety of XFP in treating urticaria. CONCLUSION: The conclusion of our systematic review will provide evidence to judge whether XFP is an effective intervention for patient with urticaria. This systematic review will be disseminated in peer-reviewed publications. The results of the study will provide evidence concerning the efficacy and safety of Xiaofeng Powder (xiao feng san) in treating urticaria. PROSPERO REGISTRATION NUMBER: PROSPERO CRD 42018087260.

Emodin Alleviates Liver Fibrosis of Mice by Reducing Infiltration of Gr1hi Monocytes.

Emodin, as a major active component of Rheum palmatum L. and Polygonum cuspidatum, has been reported to have antifibrotic effect. However, the mechanism of emodin on antifibrotic effect for liver fibrosis was still obscure. In the present study, we aimed to investigate whether emodin can alleviate carbon tetrachloride- (CCl4-) induced liver fibrosis through reducing infiltration of Gr1hi monocytes. Liver fibrosis was induced by intraperitoneal CCl4 injection in mice. Mice in the emodin group received emodin treatment by gavage. Pretreatment with emodin significantly protected mice from liver inflammation and fibrosis revealed by the decreased elevation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as reduced hepatic necrosis and fibrosis by analysis of hematoxylin-eosin (HE) staining, Masson staining, α-smooth muscle actin (α-SMA), and collagen-I immunohistochemistry staining. Further, compared to CCl4 group, mice in the emodin group showed significantly less intrahepatic infiltration of Gr1hi monocytes. Moreover, emodin significantly inhibited hepatic expression of interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), transforming growth factor-ß1 (TGF-ß1), granulin (GRN), monocyte chemoattractant protein 1 (MCP-1), and chemokine ligand 7 (CCL7), which was in line with the decreased numbers of intrahepatic Gr1hi monocytes. In conclusion, emodin can alleviate the degree of liver fibrosis by reducing infiltration of Gr1hi monocytes. These results suggest that emodin is a promising candidate to prevent and treat liver fibrosis.

Curcumin protects against liver fibrosis by attenuating infiltration of Gr1hi monocytes through inhibition of monocyte chemoattractant protein-1.

BACKGROUND AND AIMS: Liver fibrosis is concomitant with monocyte infiltration, which has been highlighted as novel therapeutic targets for chronic liver diseases. We aimed to investigate whether curcumin might protect the liver from carbon tetrachloride (CCl4)-induced fibrosis by attenuating the recruitment of Gr1hi monocytes through inhibition of monocyte chemoattractant protein-1 (MCP-1). METHODS: Mice were intraperitoneally injected with CCl4 to induce liver fibrosis. Curcumin was orally administrated to mice. Hepatic inflammation and fibrosis were evaluated by analysis of liver function and hepatic histopathology. Infiltration of the Gr1hi monocytes was assessed by flow cytometry and immunohistochemistry. Moreover, mRNA expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1ß, and transforming growth factor (TGF)-ß1 were determined by real time PCR. Hepatic expression of MCP-1 was determined by real time PCR and immunohistochemistry. RESULTS: Curcumin significantly attenuated inflammation and fibrosis, as revealed by histological and biochemical analysis. The intrahepatic infiltration of Gr1hi monocytes was attenuated by curcumin administration. T cells, NK cells, NKT cells, and dendritic cells were not affected by curcumin. Curcumin significantly reduced the expression of TNF-α and TGF-ß1, which is in line with the decreased numbers of intrahepatic Gr1hi monocytes. Intrahepatic MCP-1 expression of CCl4-challenged mice was inhibited by curcumin. CONCLUSIONS: The anti-inflammatory and antifibrotic effects of curcumin could be contributed to its prevention of Gr1hi monocyte infiltration into the injured livers through inhibition of MCP-1. These new findings extend our understanding on the mechanisms of the anti-inflammatory and antifibrotic effects of curcumin.

Schistosoma japonicum calcium-binding tegumental protein SjTP22.4 immunization confers praziquantel schistosomulumicide and antifecundity effect in mice.

A family of platyhelminth tegument-specific proteins comprising of one or two calcium ion binding EF-hand and a dynein light chain-like domain, termed tegumental proteins, are considered as candidates of vaccine. In this study, we cloned and characterized SjTP22.4, a novel membrane-anchored tegumental protein in Schistosoma japonicum with theoretic MW of 22.4. The recombinant SjTP22.4 could be recognized by S. japonicum infected sera. Immunofluorescence revealed that this protein is not only located on the surface of tegument of adult and schistosomulum and cercaria, but also in the parenchymatous tissues and intestinal epithelium. Circular dichroism (CD) measurement demonstrated rSjTP22.4 had Ca(2+)-binding ability. The rSjTP22.4 vaccination without adjuvants produced comparable high level of antibody with that of immunization with adjuvants together indicated it was an antigen of strong antigenicity. The level of IgG1 is much higher than that of IgG2a and IgE is nearly negative in S. japonicum-infected and rSjTP22.4 immunized mice. In cercaria challenge experiment, mice vaccinated with SjTP22.4 showed no reduction in adult burden and egg production, comparing with the control mice, but 41% decrease in egg mature rate and 32% reduction in liver egg granuloma area. However, the SjTP22.4 immunized mice that received praziquantel treatment at 10d post infection caused 26% reduction in adult burden and 53% decrease in egg mature rate, comparing with the control mice only received praziquantel treatment. In conclusion, SjTP22.4 is a valuable vaccine candidate for S. japonicum of anti-pathogenesis and anti-transmission effect and plays a synergetic role in praziquantel to kill schistosomulum.