System-Pharmacology Dissection of Traditional Chinese herbs SINI Decoction for Treatment of Cardiovascular Diseases.

Abstract: Cardiovascular diseases (CVDs) are leading causes of death in the world, owing to noticeable incidence and mortality. Traditional Chinese Medicine (TCM) SINI Decoction (SND) is used to prevent and treat CVDs, which has attracted extensive attention for its moderate and little side effects. However, the involved molecular mechanisms are exceedingly complicated and remain unclear. Systems pharmacology, as a novel approach that integrates systems biology and pharmacology plays a significant role in investigating the molecular mechanism of TCM. In systems pharmacology approach, we use to systematically uncover the mechanisms of action in Chinese medicinal formula SND as an effective treatment for CVDs, which mainly includes:1) molecular database building; 2) ADME evaluation; 3) target-fishing 4) network construction and analysis. The results show that 78 underlying valid ingredients and their corresponding 71 direct targets of SND were obtained. And SND take part in cardiomyocyte protection, blood pressure regulation, and lipid regulation module in treatment of CVDs by cooperative way. Systems pharmacology as an emerging field that investigates the molecular mechanisms of TCM through pharmacokinetic evaluation target prediction, and pathway analysis, which will facilitate the development of traditional Chinese herbs in modern medicine.

[Response of volatiles of Artemisiae Argyi Folium to climatic factors].

Combined headspace solid-phase microextraction with gas chromatography-mass spectrometry (HS-SPME-GC-MS), this paper studied the volatiles of Artemisiae Argyi Folium that respectively collected from Kongdong, Huanglong, Ankang, Qichun and Tongbai，and then explored the influence of climatic factors on the main components of the volatiles. The results showed that 54, 52, 57, 47 and 55 compounds were respectively identified from the leaves from five regions, mainly consist of monoterpenes, sesquiterpenoids and their oxygen-containing derivatives, a few aldehydes, alkanes and benzenes. It is different that the volatiles of Artemisiae Argyi Folium that collected in different habitats, neither composition nor content. There are total 18 kinds of compounds that exist in all five leaves. Eucalyptol, camphor, terpinen-4-ol, eugenol and caryophyllene could be considered to be the main conponents because of their efficacy or toxicity and relatively high content. Grey correlation analysis shows that the annual lowest temperature and annual sunshine duration have a great influence on the content of eucalyptol and camphor; the effect of annual precipitation and annual accumulated temperature on the content of terpinen-4-ol and caryophyllene was relatively significant; while the annual sunshine duration and the annual highest temperature are the main influencing factors for eugenol. The research suggests that long-day and extreme temperature may be more conductive to the accumulation of volatiles. According to the laws of effect of climatic factors on volatiles, we can optimize the cultivation conditions and select suitable locality to improve the quality of Artemisiae Argyi Folium.

[Screening of different AFLP fragments between near-isogenic lines of male sterile and fertile Salvia miltiorrhiza and their comparison analysis].

There is distinctive advantage of using male sterile lines to breed new cultivar and produce hybrids, when compared with general breeding method on yield and quality. In our previous work, near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained through continuous hybridization in many years. In this investigation, 378 primer combination were screened by using AFLP and BSA technique, in which 26 markers amplified from seven primers were found to tightly link to male sterile gene. Based on these markers, two linkage genetic maps were constructed. A 2 027,2 028 bp fragment was amplifed from NILs of fertile and sterile S. miltiorrhiza, respectively, using genome walking technique and previous E11/M4-208 marker as template. Four base mutations were found in intron when comparing both fragments. Among all different markers between NILs of male sterile and fertile S. miltiorrhiza, four was found to have 100% identities to chromosome 1, 3 and 5 of Arabidopsis, namely, E01/M09-418, E05/M13-308, E05/M04-750 and E01/M01-204. The E01/M09-418 marker was very close to male sterile gene of S. miltiorrhiza with distance of 2.1 cM, which also had 100% identities to male sterile gene MS2 in Arabidopsis. Both were distributed in chromosome 3 of Arabidopsis. The 2 028 bp fragment also had 100% identities to MS2 gene. Another E05/M04-750 marker that had 100% identities to chromosome 5 of Arabidopsis was found to have high identities to POP085-M05 gene of poplars and low affinity calcium antiporter CAX2 of Arabidopsis with very low E-value. The constructed genetic map and differential fragments with potential functions found in this study provide a solid foundation to lock male sterile genes in S. miltiorrhiza genome and to discover their functions.

Identification of Amplified Fragment Length Polymorphism (AFLP) Markers Tightly Associated with Drought Stress Gene in Male Sterile and Fertile Salvia miltiorrhiza Bunge.

Consistent grain yield in drought environment has attracted wide attention due to global climate change. However, the important drought-related traits/genes in crops have been rarely reported. Many near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained in our previous work through testcross and backcross in continuous field experiments conducted in 2006-2009. Both segregating sterile and fertile populations were subjected to bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) with 384 and 170 primer combinations, respectively. One out of 14 AFLP markers (E9/M3246) was identified in treated fertile population as tightly linked to the drought stress gene with a recombination frequency of 6.98% and at a distance of 7.02 cM. One of 15 other markers (E2/M5357) was identified in a treated sterile population that is closely associated with the drought stress gene. It had a recombination frequency of 4.65% and at a distance of 4.66 cM. Interestingly, the E9/M3246 fragment was found to be identical to another AFLP fragment E11/M4208 that was tightly linked to the male sterile gene of S. miltiorrhiza with 95% identity and e-value 4 × 10-93. Blastn analysis suggested that the drought stress gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1-5.

The Optimal Time of Ovarian Reserve Recovery After Laparoscopic Unilateral Ovarian Non-Endometriotic Cystectomy.

Background: Laparoscopic ovarian cystectomy is established as the standard surgical approach for the treatment of benign ovarian cysts. However, previous studies have shown that potential fertility can be directly impaired by laparoscopic ovarian cystectomy, diminished ovarian reserve (DOR), and even premature ovarian failure. Therefore, fertility-preserving interventions are required for benign gynecologic diseases. However, there are still little data on the time period required for recovery of ovarian reserve after the laparoscopic unilateral ovarian cystectomy, which is very important for the individualization of treatment protocols. This study aimed at investigating the time needed for the ovarian reserve to recover after laparoscopic unilateral ovarian non-endometriotic cystectomy. Materials and Methods: Sixty-seven patients with unilateral ovarian non-endometriotic cyst from Zhoupu and Punan Hospitals who underwent laparoscopic unilateral ovarian cystectomy were recruited as a postoperative observation group (POG). Also, 69 healthy age-matched women without ovarian cyst who did not undergo surgery were recruited as a referent group (RFG). Ovarian reserve with the serum anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), estradiol (E2) levels, ovarian arterial resistance index (OARI), and antral follicle counts (AFCs) were measured on the third to fifth days of the same menstrual cycle. A postoperative 6-month follow-up of cases was performed. Results: Compared with RFG, AFC of cyst side in the POG group showed no difference in the first, third, and sixth postoperative month (F = 0.03, F = 0.02, F = 0.55, respectively; p = 0.873, p = 0.878, p = 0.460, respectively). The OARI of cyst side in the POG group revealed no differences in the first, third, and sixth postoperative month (F = 0.73, F = 3.57, F = 1.75, respectively; p = 0.395, p = 0.061, p = 0.701, respectively). In the first month, the postoperative AMH levels significantly declined, reaching 1.88 ng/ml [interquartile range (IQR): 1.61-2.16 ng/ml] in POG and 2.57 ng/ml (IQR: 2.32-2.83 ng/ml) in RFG (F = 13.43, p = 0.000). For the data of AMH levels stratified by age, the same trend was observed between less than 25 and more than 26 years old. At this same time interval, the postoperative rate of decline was significantly lower compared to the preoperative one in POG (32.75%). The same trend was observed between the POG and RFG groups (26.67%). Conclusions: The optimal time for recovery of ovarian reserve after laparoscopic unilateral ovarian cystectomy is estimated to be 6 months.

iTRAQ-based proteomic analysis reveals several key metabolic pathways associated with male sterility in Salvia miltiorrhiza.

Male sterility is a common phenomenon in flowering plants, and it has been widely used in hybrid seed production in a number of economically important crops. In 2002, our team discovered a natural male sterile mutant of Salvia miltiorrhiza. It provided us with the possibility of obtaining stable and controllable quality. To study the molecular mechanism of male sterility in S. miltiorrhiza, we generated proteomic profiles comparing the male sterile mutant type (MT) and wild type (WT) using iTRAQ sequencing. We found a total of 639 differential abundant proteins (DAPs) between MT and WT buds. The DAPs associated with male sterility were mainly involved in (1) carbohydrate and energy metabolism, and (2) protein synthesis and degradation. Based on a comparison between the protein expression profiles of MT and WT, we elucidated a potential protein interaction network involved in male sterility. These results provide new potential biomarkers and insights into the molecular mechanism of male sterility in S. miltiorrhiza.

Clarifying of the potential mechanism of Sinisan formula for treatment of chronic hepatitis by systems pharmacology method.

Chronic hepatitis is a general designation class of diseases, which results in different degrees of liver necrosis and inflammatory reaction, followed by liver fibrosis, may eventually develop into cirrhosis. However, the molecular pathogenesis of chronic hepatitis is too complex to elucidate. Herbal medicines, featured with multiple targets and compounds, have long displayed therapeutic effect in treating chronic hepatitis, though their molecular mechanisms of contribution remain indistinct. This research utilized the network pharmacology to confirm the molecular pathogenesis of chronic hepatitis through providing a comprehensive analysis of active chemicals, drug targets and pathways' interaction of Sinisan formula for treating chronic hepatitis. The outcomes showed that 80 active ingredients of Sinisan formula interacting with 91 therapeutic proteins were authenticated. Sinisan formula potentially participates in immune modulation, anti-inflammatory and antiviral activities, even has regulating effects on lipid metabolism. These mechanisms directly or indirectly are involved in curing chronic hepatitis by an interaction way. The network pharmacology based analysis demonstrated that Sinisan has multi-scale curative activity in regulating chronic hepatitis related biological processes, which provides a new potential way for modern medicine in the treatment of chronic diseases.

[Preliminary study on the characteristics of Bupleurum chinense in nutrition uptake].

It was studied that the characteristics of Bupleurum chinense DC. in nutrition uptake based on potted and field experiment to provide a properly fertilizer application measure. The result showed that the absorption amounts of N, P and K were in the order of K > N > P under field condition. For one-year-old B. chinesnse DC., the average N, P2O5 and K2O absorption per 100 kg dry matter of root were 11.77 kg, 2.43 kg, 14.07 kg respectively; for two-year-old, the average N, P2O5 and K2O absorption amount per 100 kg dry matter of root were 12.11 kg, 2.74 kg, 18.39 kg respectively. The total absorption amount of N, P2O5 and K2O were lower in the first year, while the percentage of content was higher, that was to say, the demand for N, P and K was urgent in the first year. The accumulation peak of N, P2O5 and K2O occurred regreening to later elongation stage in the second year. The ratio of N, P and K were differential in different growth stages.

[Preliminary study on N, P, K fertilizer to control of root rot of Bupleurum chinense].

The application of N and K fertillizer could improve the sensibility of Bupleurvum chinense DC. to Root Rot, while large application of P fertilizer could decrase the sensibility. The fertilizer measure which could obtain highest yield but could not increase its disease resistense. To protect Bupleurum chinense against root rot, more phosphorous fertilizer, certain nitrogen and potassium ferilizer should be applied in early elongation stage in the second growing year.

[Microecological mechanisms of red-leaf disease occurrence in Salvia miltiorrhiza Bge].

A comparative study was made on the nutrient content in rhizosphere soil and the microflora in rhizosphere soil and on rhizoplane of healthy and red-leaf diseased Salvia miltiorrhiza plants, aimed to approach the microecological mechanisms of red-leaf disease occurrence in S. miltiorrhiza. The N, P, K, and Mn contents in the diseased plant leaves were significantly lower than those in the healthy plant leaves (P < 0.05). No significant difference was observed in the available P content in the rhizosphere soils of diseased and healthy S. miltiorrhiza, but the available N and K contents were significantly higher (P < 0.05) in the rhizosphere soil of diseased S. miltiorrhiza. These results indicated that the red-leaf disease occurrence in S. miltiorrhiza was related to plant P deficiency, but the lack of P in the plants was not caused by the insufficient soil P supply. As compared with those in healthy S. miltiorrhiza rhizosphere, the bacterial number in diseased S. miltiorrhiza rhizosphere soil decreased by 41.3% , while the fungal and actinomycetes numbers increased by 156.6% and 189.5% (P < 0.05), respectively. Similar variations in the numbers of bacteria, fungi, and actinomycetes were observed on diseased S. miltiorrhiza rhizoplane. In the rhizosphere soil and on the rhizoplane of diseased S. miltiorrhiza, the predominant microbial species that might be harmful included six fungi (Fusarium solani, Myrothecium roridum, F. tricinctum, Aspergillus calidoustus, F. oxysporum, and Dothideomycetes sp.), four actinomycetes (Streptomyces lateritius, Lentzea waywayandensis, S. stelliscabiei and S. collinus), and two bacteria (Bacillus aryabhattai and Piscinibacter aquaticus). These predominant soil microbes likely caused plant P deficiency via negatively affecting the growth of roots and their absorption of soil nutrients. It was suggested that the red-leaf disease occurrence in S. miltiorrhiza was closely related to the plant P deficiency caused by the abnormality of soil microflora in the rhizosphere soil and on the rhizoplane of S. miltiorrhiza.

A dominant gene for male sterility in Salvia miltiorrhiza Bunge.

A natural male sterile mutant of Salvia miltiorrhiza (Labiatae, Sh-B) was found during field survey in 2002. Our objective was to analyze its genetic mechanism for producing F1 hybrid seeds and to develop a molecular marker linked to male sterile gene for selection of a hybrid parent line. The segregation ratios of male sterile plants to fertile plants in the progenies of both testcross and backcross were 1:1 in continuous experiments conducted in 2006-2009. The male sterile Sh-B was heterozygous (Msms). The male sterile plants could capture most pollen (2 granule/cm(2)·24 h) with row ratio (female:male 2:1) within 45-cm distance and harvest the largest amount of 6495 g hybrid seeds per hectare. We also developed DNA markers linked to the male sterile gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 128 primer combinations. One out of fourteen AFLP markers (E11/M4208) was identified as tightly linked to the dominant male sterile gene with a recombination frequency of 6.85% and at a distance of 6.89 cM. This marker could be converted to PCR-based assay for large-scale selection of fertile plants in MAS (marker-assisted selection) at the seedling stage. Blastn analysis indicated that the male sterile gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1-5, and was more likely to encode S-adenosylmethionine-dependent methyltransferase, in which DNA methylation regulated the development of plant gametogenesis.