Objective To understand the research status and development trend of TCM in the treatment of lower limb arteriosclerosis obliterans(LLASO);To provide reference for related research.Methods The literature on the treatment of LLASO by TCM was retrieved from CNKI,Wanfang Data and VIP from September 18,1992 to September 18,2022.NoteExpress 3.6.0 was used to manage and remove repetition.Excel 2019 was used to analyze the trend of the number of articles published.VOSviewer 1.6.18.0 and CiteSpace 6.1.R6 were used to analyze the co-occurrence,clustering,timeline and emergence of the included authors,institutions and keywords,and knowledge graph was drawn.Results A total of 1 095 articles were included,and the number of articles published showed an overall upward trend;1 915 authors were involved,and the author with the most articles was Chen Bainan,forming research teams with Xi Jiuyi,Cao Yemin and Ge Jianli as the cores respectively;the main research institutions were Shandong University of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Heilongjiang University of Chinese Medicine and so on,but there was little cooperation among them.High-frequency keywords included arteriosclerosis,diabetes,clinical efficacy,gangrene,hemorheology,etc.,forming 13 clusters such as qi deficiency and blood stasis,Simiao Yong'an Decoction,angiogenesis and so on.The focus of research was external treatment of TCM,clinical efficacy,blood stasis syndrome,medication law,Buyang Huanwu Decoction.Conclusion The research heat of TCM in the treatment of LLASO has gradually increased,and the modern research in this field from 1992 to 2022,such as gene detection and molecular biology based on big data platform,is the research hotspot in this field.The development trend is the combination of internal treatment and external treatment of TCM,as well as the interdisciplinary research of integrated traditional Chinese and Western medicine combined with interventional therapy and other techniques.

OBJECTIVE To establish the quality standard of Clinopodium gracile. METHODS Ten batches of C. gracile were collected to perform appearance and property identification， microscopic identification and thin layer chromatography （TLC） identification. Moisture， total ash， acid-insoluble ash and dilute ethanol extract were detected， and the content of rosmarinic acid was determined by HPLC. RESULTS The stem of C. gracile was slender， square columnar， covered by white fluff， the surface was grayish green or greenish brown； epidermal cells， non-glandular hairs， cortical cells and so on were seen in the cross section of the stem. Non-glandular hairs， ducts， wood fibers， mesophyll cells and so on could be seen in the powder. Results of TLC identification showed that there were spots of the same color in the chromatographic position corresponding to the chromatographic position of buddlejasaponin Ⅳb control. The contents of water， total ash， acid-insoluble ash， dilute ethanol extract and rosmarinic acid in 10 batches of samples were 8.69%-12.33%， 5.96%-13.33%， 0.14%-3.29%， 18.57%-32.61%， 0.35%-0.82%， respectively. The average values were 10.10%， 9.73%， 1.06%， 23.54% and 0.56%， respectively. CONCLUSIONS The established method can be used for quality control of C. gracile. It is preliminarily proposed that the ash content in the herb should not exceed 12.0%， the total ash content should not exceed 12.0%， the acid-insoluble ash content should not exceed 1.5%， the dilute ethanol extract should not be less than 18.0%， and the rosmarinic acid content should not be less than 0.45%.

ObjectiveTo study the accumulation and absorption of dry matter and nutrients in Curcuma phaeocaulis from Sichuan， the origin of Curcumae Radix （tuber） and Curcumae Rhizoma （rhizome）， to explore the growth and development laws and nutrient demand characteristics of the medicinal species， and thus to lay a theoretical basis for rational fertilization. MethodThe plant growth indexes， dry matter accumulation， and nutrient content of C. phaeocaulis at different growth stages in Sichuan were measured and analyzed. ResultThis medicinal species featured the dominant growth of aboveground leaves and stems before October and growth of underground part （particularly the rhizomes and tubers） from October. During the whole growth period， the accumulation of nitrogen， phosphorus， potassium， calcium， magnesium， iron， manganese， zinc， and copper per plant was 2 450.31， 907.09， 3 171.18， 625.94， 493.38， 14.53， 2.24 ， 2.93， 0.46 mg， respectively， with the order of potassium > nitrogen > phosphorus > calcium > magnesium > iron > zinc > manganese > copper. ConclusionThe species needs sufficient potassium and nitrogen， appropriate amount of phosphorus， calcium， and magnesium， a small amount of iron， and very little zinc， manganese， and copper for growth， and potassium is particularly important. Nitrogen， phosphorus， and potassium fertilizers supply macroelements and nitrogen fertilizer should be supplemented at seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome. Phosphorus and potassium fertilizers should be applied at seedling stage， tuber expansion stage， secondary expansion stage of rhizome， and dry matter accumulation stage of tuber and rhizome. At seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome， a variety of medium and trace elements-containing foliar fertilizers should be used.

Chinese medicinal material from Fritillaria， Beimu in Chinese， is a commonly used antitussive and expectorant traditional Chinese herbal medicine， with the significant functions of clearing heat and moistening lung，resolving phlegm and relieving cough. Five kinds of Fritillaria were recorded in the 2020 edition of Chinese Pharmacopoeia， including Fritillariae Cirrhosae Bulbus， Fritillariae Ussuriensis Bulbus， Fritillariae Thunbergii Bulbus， Fritillariae Hupehensis Bulbus and Fritillariae Pallidiflorae Bulbus. At present， the reports on Fritillaria mainly focus on the pharmacological effect， chemical composition， identification of authenticity and other aspects， while there were few reports on harvesting and primary processing of original medicinal materials. Fritillaria medicines were perennial medicinal plant with various and complex varieties， their quality and curative effect were greatly affected by harvesting and processing in producing area. The processing method differed according to its variety. Fritillariae Cirrhosae Bulbus mainly from western Sichuan plateau， Fritillariae Pallidiflorae Bulbus from Xinjiang and Fritillariae Ussuriensis Bulbus from Northeast China were mostly harvested from June to July and sun dried directly or dried. But Fritillariae Thunbergii Bulbus and Fritillariae Hupehensis Bulbus from Yangtze River basin were harvested when the plants wilted at the beginning of summer， and auxiliary materials such as shell powder and lime must be added during the processing. At present， the drying methods of Fritillaria were still traditional， which is not suitable for large-scale production of cultivated products. Therefore， it is urgent to find a scientific， reasonable and efficient processing methods. Aimed at providing references for standardization production of Fritillaria， this paper made a textual research on the ancient and modern herbal literature， the Chinese Pharmacopoeia and other medicinal standards， combined with modern literature， the harvesting and processing methods of Fritillaria were sorted out and prospected.

Objective:To learn the depression status of the hospitalized pregnant women in labor and its related influencing factors,and to provide the theoretical basis for exploring depression intervention of the pregnant women in labor.Methods:Self-Rating Depression Scale was used for investigating the depression status of 203 hospitalized pregnant women in labor who were in hospital from November 2014 to May 2015,and the impacts of demographic characteristics,maternal history and physical condition and health care on depression status of hospitalized pregnant women in labor before pregnancy and during pregnancy were analyzed using Logistic regression.Results:Fifty-three people with depressive tendencies were detected among pregnant women in labor,and the positive rate was 26.1％.The differences in positive rates of depression tendency of the pregnant women between less than 30 years,30-39 years and 40 years and above had no statistical significance (P＞0.05).Univariate Logistic analysis showed that the differences of occupation,overweight or obesity,low-dose folic acid supplements before pregnancy were statistically significant (P＜0.01).Multivariate Logistic regression analysis showed that risk factor for pregnant women was overweight or obesity pre-pregnancy,and the risk was 2.335 times to non-overweight or non-obesity pregnancy women (OR=2.335,95％CI:1.101-4.954,P＜0.05).Conclusion:The pregnant women have the depression tendency,especially the pregnant women with high body mass index before pregnancy.The knowledge of psychological health education should be provided for them.

The afsRS(cla) global regulatory genes from Streptomyces clavuligerus activate the production of two antibiotics in Streptomyces lividans. In this study, we gained an increase of 38% in the production of natamycin (3.56 g/L) in an industrial strain Streptomyces gilvosporeus TZ1401 through the integration of pHL851 that bears the afsRS(cla) global regulatory genes into its genome. We discovered by quantitive real-time reverse transcription PCR (qRT-PCR) that the expression of 6 genes of the natamycin biosynthetic gene cluster were improved from 1.9 to 2.7 times. This suggests that afsRS(cla) improve the production of natamycin through increased transcription. This study provides a good example for applying afsRS(cla) in high yield breeding of industrial antibiotic producers.
Anti-Bacterial Agents ; biosynthesis ; Genes, Regulator ; Industrial Microbiology ; Multigene Family ; Natamycin ; biosynthesis ; Streptomyces ; genetics

AIM:To investigate the potential effect of all-trans retinoic acid (ATRA) on the proliferation of human breast cancer MCF-7 cells and its underlying mechanism.METHODS:Human breast cancer MCF-7 cells were in-cubated in DMEM supplemented with 10%fetal bovine serum and 1%penicillin/streptomycin.Western blot was performed to detect the protein expression and its phosphorylation.MTT assay and bromodeoxyuridine ( BrdU) incorporation, and TUNEL staining were carried out to determine the cell proliferation and apoptosis, respectively.Lentivirus carrying shRNA sequences targeting the gene of docking protein 1 ( DOK1 ) was used to silence DOK1 expression.The activity of peroxi-some proliferator-activated receptor gamma ( PPARγ) was measured using a PPARγtranscription factor assay kit.RE-SULTS:ATRA treatment suppressed the proliferation and promoted the apoptosis of MCF-7 cells.ATRA was also found to induce DOK1 expression in a time-dependent manner.Silence of DOK1 mitigated anti-cancer effect of ATRA evidenced by recovered the cell proliferation and reduced the cell apoptosis.In addition, DOK1 knockdown inhibited PPARγexpression and activity.Furthermore, inhibition of PPARγwith its specific inhibitor GW9662 attenuated impacts of ATRA on the cell proliferation and apoptosis.CONCLUSION: ATRA suppresses MCF-7 cell survival through inhibiting cell proliferation and promoting cell apoptosis, which is mediated by DOK1-activated PPARγ.

AIM: To investigate the role of ubiquitin-proteasome system ( UPS) in adipocyte differentiation. METHODS:Differentiation of 3T3-L1 preadipocytes into adipocytes was induced by treatment with insulin, 3-isobutyl-1-methylxanthine and dexamethasone.Western blot and immunoprecipitation were performed to detect the protein abundances and association, respectively.Oil red O staining was used to determine the intracellular lipid of 3T3-L1 adipocytes.The levels of mRNA were measured by reverse transcription polymerase chain reaction ( RT-PCR) .RESULTS:UPS inhibitor bortezomib (BZM) suppressed the differentiation of 3T3-L1 pre-adipocytes, evidenced by reduced intracellular content of triglyceride, and decreased mRNA expression of adipogenic marker proteins such as adiponectin and adipocyte protein 2.In contrast, administration of sildenafil (SDN), an activator of protein kinase G which was also found to activate UPS, promo-ted adipocyte differentiation.In addition, BZM treatment decreased the expression of heat shock protein 90 (HSP90) and peroxisome proliferator-activated receptor gamma ( PPARγ) in the soluble fraction and reduced association of HSP90 and PPARγ.Furthermore, HSP90-specific N-terminal inhibitor geldanamic mitigated SDN-induced increase in PPARγlevel and 3T3-L1 cell differentiation.CONCLUSION:UPS modulates HSP90-dependent PPARγstability, thus leading to pro-motion of adipocyte differentiation.

Objective To investigate the survival of advanced stage periampullary carcinoma and its prognostic factors.Methods The clinical data and follow up of 134 patients with advanced stage periampullary carcinoma admitted to our hospital between January 2007 and December 2010 were retrospectively collected and analyzed.Results Of 134 patients,there were 83 males and 51 females.The mean age±S.D.was 62.49±11.41 (28-83).For the 86 patients who underwent surgery,32 had extensive peritoneal metastases on surgical exploration,27 had metastases/involvement of the liver or stomach,and 27 had major vessel infiltration.For these 86 patients,55 received surgical exploration,whereas 31 had surgical exploration plus bypass anastomosis. 45 patients received chemotherapy,while 67 received chemotherapy plus radiotherapy.The improvements in quality of life and in clinical response rate after concurrent chemoradiotherapy or chemotherapy alone were 73.1％ and 57.8％,respectively.The overall survival was 14.27±1.06 months.The median survival time was 11 (8.95±13.05) months.The 1- and 3-year survival rates were 43％ and 10％,respectively.Karnofsky performance status,and the presence or absence of jaundice co-related with poor prognosis on single factor analysis.The Karnofsky performance status was an independent survival predictor on multifactor analysis.Conclusions The prognosis of patients with advanced stage periampullary carcinoma was poor.Chemotherapy with or without radiotherapy had similar impact on overall survival,Karnofsky performance status was an independent survival predictor.

Tunicamycin, a potent reversible translocase I inhibitor, is produced by several Actinomycetes species. The tunicamycin structure is highly unusual, and contains an 11-carbon dialdose sugar and an α, β-1″,11'-glycosidic linkage. Here we report the identification of a gene cluster essential for tunicamycin biosynthesis by high-throughput heterologous expression (HHE) strategy combined with a bioassay. Introduction of the genes into heterologous non-producing Streptomyces hosts results in production of tunicamycin by these strains, demonstrating the role of the genes for the biosynthesis of tunicamycins. Gene disruption experiments coupled with bioinformatic analysis revealed that the tunicamycin gene cluster is minimally composed of 12 genes (tunA-tunL). Amongst these is a putative radical SAM enzyme (Tun B) with a potentially unique role in biosynthetic carbon-carbon bond formation. Hence, a seven-step novel pathway is proposed for tunicamycin biosynthesis. Moreover, two gene clusters for the potential biosynthesis of tunicamycin-like antibiotics were also identified in Streptomyces clavuligerus ATCC 27064 and Actinosynnema mirums DSM 43827. These data provide clarification of the novel mechanisms for tunicamycin biosynthesis, and for the generation of new-designer tunicamycin analogs with selective/enhanced bioactivity via combinatorial biosynthesis strategies.
Actinobacteria ; enzymology ; genetics ; Base Sequence ; Biological Assay ; Carbohydrate Sequence ; Carbohydrates ; biosynthesis ; genetics ; Cloning, Molecular ; Gene Deletion ; Gene Library ; High-Throughput Screening Assays ; Molecular Sequence Data ; Multigene Family ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; Streptomyces ; enzymology ; genetics ; Tunicamycin ; biosynthesis ; chemistry ; genetics