Integration of network pharmacology and bone marrow mesenchymal stem cells experimental research to reveal the molecular mechanisms for Hai Honghua medicinal liquor against osteoporosis

Background:Hai Honghua medicinal liquor(HHML)formula has been used in clinical practice for a long time,mainly for the treatment of freshly closed fractures,to promote osteogenesis,to increase bone mass,and thus to promote fracture healing.However,the underlying mechanisms of HHML in the treatment of osteoporosis(OP)are still unclear.Methods:Firstly,Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform and The Encyclopedia of Traditional Chinese Medicine were used to screen the targets of the active compounds of HHML.At the same time,OP targets were identified through GeneCards,Online Mendelian Inheritance in Man,DisGeNET,Therapeutic Target Database,Comparative Toxicogenomics Database and Human Phenotype Ontology databases.Next,protein-protein interaction and pathway networks were constructed for compound-disease common targets,and core targets and compounds were screened for molecular docking.Furthermore,rat bone marrow mesenchymal stem cells were extracted as model cells,and the osteogenic effects of HHML were verified via in vitro experiments.Results:Total of 343 common targets of HHML-OP and the top 10 target proteins in the protein-protein interaction network are TP53,AKT1,STAT3,HSP90AA1,ESR1,TNF,IL6,MAPK1,MAPK3 and MAPK8.Enrichment analysis yielded that the key molecular pathway was the PI3K/Akt signaling pathway.Molecular docking analysis showed that baicalein,erysodienone and naringenin docked with the target proteins AKT1,STAT3 and TP53,respectively,with low binding energy and high affinity.In addition,In vitro experiments demonstrated that HHML promoted the proliferation of bone marrow mesenchymal stem cells;compared with the control group,HHML-treated cells showed enhanced alkaline phosphatase staining,promoted the expression of OCN,RUNX2,BSP,and COL1 mRNAs as well as the expression of PI3K and AKT phosphorylated proteins.Secondly,the expression of target proteins revealed that HHML promoted the phosphorylation of STAT3 protein and inhibited the expression of P53.Conclusions:Our study investigated that HHML treatment with OP promotes bone formation possibly through activation of the PI3K/Akt signaling pathway and may involve STAT3 and TP53 target interactions.

The mechanistic study on Wa medicine Niang-Mu-Liang medicinal liquor mitigating diabetes mellitus erectile dysfunction in rats by inhibiting ferroptosis

Background:This study aims to investigate the therapeutic effect of Wa medicine Niang-Mu-Liang medicinal liquor(NML)on rats with diabetes mellitus erectile dysfunction(DMED)and its impact on the ferroptosis signaling pathway.Methods:Thirty Sprague-Dawley rats were randomly divided into three groups:Control,DMED,and NML.After establishing the DMED model,treatments were administered for 8 weeks.After the administration,apomorphine hydrochloride tests were conducted to measure the mass and organ index of testes and epididymides,sperm concentration and viability in each group.Penile corpus cavernosum tissues were stained with hematoxylin and eosin.Nitric oxide and cyclic guanosine monophosphate levels in the penile corpus cavernosum tissues were determined using biochemical kits and enzyme-linked immunosorbent assay,while the expression of proteins related to the ferroptosis signaling pathway was measured by Western blot.Results:Compared to the DMED group,the DMED rats treated with NML showed significantly increased erection frequency,testicular and epididymal mass and index,sperm count and viability,along with noticeable improvement in the pathological morphology of penile corpus cavernosum.The content of nitric oxide and cyclic guanosine monophosphate,and the expression of ferritin heavy chain,ferritin light chain,and glutathione peroxidase 4 proteins in penile corpus cavernosum tissue were elevated,while the expression of transferrin and STEAP3 proteins was reduced.Conclusion:NML can improve erectile function in DMED rats by inhibiting the ferroptosis signaling pathway.

Research of the molecular mechanism of Hai Honghua medicinal liquor in the treatment of fracture based on network pharmacology combined with molecular docking

Background:Hai Honghua medicinal liquor(HHML),a famous hospital formula composed of 19 traditional Chinese medicines,has been successfully applied in treating soft tissue injury,fresh closed fracture,limb dysfunction after fracture healing,shoulder,neck and leg pain,knee joint pain and other clinical multiple diseases for 30 years in clinical.However,research on the material basis of HHML for the treatment of fracture healing-related disorders is still in a gap.Therefore,it is particularly important to explore the active ingredients,core targets and potential pharmacological mechanisms of HHML to promote fracture healing.Methods:We screened the core active components of each traditional Chinese medicine in formula and its action targets through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Encyclopedia of Traditional Chinese Medicine database.The fracture related targets were retrieved from several different public databases,including GeneCards,Online Mendelian Inheritance in Man,DisGeNET and Therapeutic Target Database.Bioinformatics analysis to obtain key bioactive components,underlying targets and signaling pathways,containing the Venn diagram of the intersection with components and diseases gene targets,protein–protein interaction,as well as the Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes analysis,and finally molecular docking.Results:A total of 249 bioactive ingredients of HHML and 325 HHML-fracture-related targets were screened.The network analysis revealed that quercetin,luteolin,kaempferol,Licochalcone A,naringenin and 8-Isopentenyl-kaempferol may be potential candidate agents.Multiple targets are involved including TP53,MAPK3,STAT3,AKT1,MAPK1,HSP90AA1,ESR1 and PIK3CA may be closely linked targets.PI3K-AKT signaling pathway may play a significant role of HHML in treatment of fracture.What’s more,molecular docking suggested that 8-isopentenyl kaempferol,glycyrrhiza chalcone A,and naringenin bound to AKT1,PIK3CA,and ESR1,respectively,exhibiting lower energy and more stable characteristics.Conclusions:The findings indicate the potential active ingredients,target proteins and molecular mechanisms of HHML for the treatment of fractures to provide the exact idea for the next research on the mechanism of action of HHML formula for fracture treatment.

Mesenteric phlebosclerosis with amyloidosis in association with the long-term use of medicinal liquor: A case report

BACKGROUND Mesenteric phlebosclerosis(MP)is a rare disease of the colon.The clinical manifestations of this disease are nonspecific and it may easily be misdiagnosed.We report a case of MP with amyloidosis in the colonic vessel walls in a patient with hypertension who had been consuming Chinese medicinal liquor for 10 years.We also review the relevant literature and summarize the characteristics of MP in patients in China's Mainland.CASE SUMMARY A 64-year-old man was referred to our department from his primary hospital because of abdominal pain,diarrhea,and fever for almost 10 d.Computed tomography showed colon wall thickening,with threadlike calcifications in the mesenteric vein in the transverse colon.Colonoscopy revealed purple-blue mucosa with multiple ulcers in the ascending and transverse colon.Biopsy showed thickening and calcification of the vein walls,perivascular and mucosal collagen degeneration,and amyloidosis.The patient had been consuming Chinese medicinal liquor,mainly that made from gardenia fruit,for 10 years.Based on these results,a diagnosis of MP with amyloidosis was made.After conservative treatment,the patient’s discomfort subsided and he was followed closely.The use of Chinese herbal medicine was suspected to play a role in the pathogenesis of MP.CONCLUSION The clinical manifestations of MP are nonspecific.Recognition of its typical imaging findings,including multiple calcifications on computed tomography and purple-blue mucosal discoloration on colonoscopy,is vital.

Beijing Tong Ren Tang Medicinal Liquor Plant

The Beijing Tong Ren Tang Medicinal Liquor Plant under the China Beijing Tong Ren Tang Holdings Corporation, officially built in Tongxian County in the suburbs of Beijing in 1971, has gradually developed into the biggest producer of medicinal liquors in the country in the past 20 years, and became a State second-class enterprise in 1990. The plant is known for its

间接竞争酶联免疫吸附测定法快速筛查药酒中双氯芬酸

目的建立间接竞争酶联免疫吸附测定法快速筛查药酒中双氯芬酸(diclofenac,DCF)。方法采用活性酯法和混合酸酐法分别将DCF与载体蛋白耦合,得到DCF的免疫原和检测抗原。采用DCF-牛血清白蛋白免疫BALB/c小鼠,随后用杂交瘤等技术制备抗DCF单克隆抗体(monoclon alantibody,mAb),基于DCFmAb建立了间接竞争酶联免疫吸附测定法,对该检测方法的性能(准确度、精密度和特异性)进行鉴定。结果紫外扫描结果表明DCF已成功与载体蛋白偶联;获得最优株抗DCF的杂交瘤细胞株(4B9),其IC50值为0.61ng/mL;该检测方法在药酒中的DCF平均添加回收率为85.9%,其批间变异系数(coefficient of variation,CV)(5.3%~9.7%)均大于批内CV(4.9%~9.1%),与类似物(酮洛芬、阿司匹林、吲哚美辛、布洛芬、舒林酸、萘普生、罗非昔布)没有交叉反应。结论本研究建立的间接竞争ELISA法为DCF在药酒中的残留提供了一种新的筛查手段。

中药酒剂配合集束化护理在颈肩腰腿痛中的应用

目的探讨中药酒剂配合集束化护理在颈肩腰腿痛治疗中的应用。方法选取2020年6月—2022年6月收治的颈肩腰腿痛患者50例,随机分为两组,对照组采取基础护理及电磁治疗仪治疗,研究组在对照组基础上采取中药酒剂配合集束化护理。比较两组不同时间点疼痛程度、运动功能改善效果及护理满意度。结果两组护理后3 d、护理后1周疼痛评分均低于护理前(P<0.05),研究组护理后3 d、护理后1周疼痛评分均低于对照组(P<0.05);研究组运动功能改善优于对照组(P<0.05);研究组护理满意度96.0%(24/25)高于对照组的76.0%(19/25)(P<0.05)。结论中药酒剂配合集束化护理应用在颈肩腰腿痛患者护理中,可以明显减轻患者疼痛感,改善患者运动功能,有利于患者身体康复,患者满意度较高,值得护理临床应用。

医院制剂蛇药酒抑制眼镜蛇毒酶活性的实验研究

目的探讨医院制剂蛇药酒抗眼镜蛇毒中4种主要酶活性的效果。方法采用不同浓度蛇药酒与蛇毒共孵育作为蛇药酒组,同时设置无蛇药酒的蛇毒对照组。通过琼脂平板法比较蛇药酒组及蛇毒对照组产生透明圈的直径,以检测不同浓度蛇药酒对眼镜蛇毒中磷脂酶A_(2)(PLA_(2))、纤维蛋白原溶酶(以下简称纤溶酶)的抑制作用;分别以福林法和浊度法检测两组的蛋白水解酶和透明质酸酶活性,观察蛇药酒对眼镜蛇毒中蛋白水解酶、透明质酸酶的抑制作用。结果蛇药酒浓度为0.0008、0.004、0.02、0.1、0.5 mg/ml时,对眼镜蛇毒中的PLA_(2)有较强的抑制作用;蛇药酒浓度为1.3、13、130 mg/ml时,可明显抑制纤溶酶活性;蛇药酒浓度为0.5、2、8、32 mg/ml时,对蛇毒中蛋白水解酶抑制作用显著;蛇药酒浓度为0.0625、0.125、0.25、0.5、1 mg/ml时,对眼镜蛇毒中透明质酸酶抑制作用呈剂量依赖性。结论医院制剂蛇药酒在体外有直接抑制眼镜蛇毒中PLA_(2)、纤溶酶、蛋白水解酶及透明质酸酶的作用。

Treatment of Chinese traditional medicine wastewater in a submerged membrane bioreactor

A pilot scale test was conducted in a submerged membrane bioreactor SMBR with capacity of 10. 0 m^3/d for 120 days to treat high-strength Chinese traditional medicine wastewater. Performance of the SMBR was investigated with a sludge retention time （ TSR ） of 50 days, a hydraulic retention time （ THR ） of 8.0 h, membrane flux of 8. 0 IV（ m^2 · h） and dissolved oxygen （DO） concentration of 2. 0 - 3. 0 mg/L, respectively. It was observed that the SMBR had high capacity of COD and suspended solid （SS） removal. The influent COD concentration was fluctuated between I 000 and 5 000 mg/L, while the averaged effluent COl） concentration was only 44. 6 mg/L. The influent SS concentration was fluctuated between 1 000 and 1 600 mg/L, while little effluent SS was detected. It was found that the COD remove rate increased with mixed liquor suspended solids （MLSS） and organic loading rate （ROL）. In order to obtain good-quality effluent, the operational conditions of the SMBR were suggested as follows： the temperature was controlled above 10 ℃, MLSS about 7000 mg/L, R,L under 24. 76 kg/（ m^3 · d）, low vacuum value and constant water flux.

应用液质联用法(LC-MS/MS)快速测定药酒中11种钩吻生物碱的含量

目的建立高效液相色谱-质谱联用法(液质联用法,LC-MS/MS)测定药酒中钩吻素甲等11种钩吻生物碱的含量,以期为药酒中毒事件中样品的钩吻碱等生物碱的应急检测提供参考。方法采用LC-MS/MS法,色谱柱为Agilent ZORBAX Eclipse XDB C18柱(2.1×150 mm,3.5μm);柱温为40℃;流动相中A相为0.1%甲酸水溶液,B相为0.1%甲酸乙腈;流速为0.5 mL/min;以钩吻素甲等11种钩吻生物碱为标准对照品,同时测定药酒样品中各物质含量。结果11种钩吻生物碱在5.0~300.0 ng/mL范围内线性关系良好,相关系数为0.9981~0.9998,回收率在85.2%~98.4%之间,检出限为0.1~0.6μg/kg,定量限为0.3~2.0μg/kg。结论高效液相色谱-质谱联用法具有选择性强、灵敏度好、准确度高的优势,可用于药酒样品中生物碱的快速定性和定量。

中药酒制法的研究进展

目的综述酒制法的历史沿革和现代研究现状。方法查阅中药酒制法相关文献,并加以归纳和整理。结果酒制法始于春秋战国时期,唐代酒制法进一步发展,明代立说"酒制升提"。酒制法的现代研究包括酒制理论性研究和实验性研究。理论性研究重点讨论酒制升提的科学性与局限性。实验性研究包括酒制原理、酒制工艺规范化研究和酒制中药类饮片质量标准,其中酒制原理主要从药性、成分、药理方面说明酒制对中药作用的影响。结论中药酒制法仍有许多发展空间和展望,对于酒制机理的进一步研究也具有一定的临床价值。

响应面法优化壳聚糖澄清药酒工艺及其效果评价

以中药原酒为原料,根据Box-Behnkend试验设计原理,采用响应面分析法,建立壳聚糖澄清药酒的二次多项数学模型,并以药酒澄清度为响应值和等高线建立响应面,研究壳聚糖浓度、处理温度、处理时间对药酒澄清效果的影响。结果表明:壳聚糖澄清药酒的最佳工艺条件为澄清剂添加剂量8.0g/L、处理温度32.6℃、处理时间46.4min;与常规法澄清药酒相比,经过处理的药酒澄清度提高了13.4%。

顶空固相微萃取-气相色谱-质谱法测定药香型白酒中萜烯类化合物

应用顶空固相微萃取与气相色谱-质谱联用技术定量药香型白酒中的痕量萜烯类化合物。实验考察萃取头、样品酒精度、萃取温度和萃取时间对萜烯类化合物萃取效果的影响。最优顶空固相微萃取条件为将酒样稀释到酒精度10%,采用50/30μm碳分子筛/聚二乙烯苯/聚二甲基硅氧烷(DVB/CAR/PDMS)萃取头在60℃条件下萃取45min。该方法具有良好的线性,所有化合物的线性相关系数良好,R2≥0.9921,检测限为0.01～3.62μg/L,定量限为0.03～12.05μg/L,相对标准偏差小于15%,加标回收率为81.01%～118.94%。用该方法对药香型白酒的成品酒、原酒中的萜烯类化合物进行测定,共定量41种挥发萜烯类化合物,结果发现大曲香醅原酒中的萜烯总含量要多于小曲酒醅原酒中萜烯总含量,D-樟脑等17萜烯只在大曲香醅原酒中检测到,在小曲酒醅原酒中没有检测到,排除蒸馏时酸、热等带来的影响,推测这17种萜烯主要来源于大曲香醅,相反卡拉烯只在小曲酒醅原酒中检测到,推测其来源于小曲酒醅。顶空固相微萃取适合药香型白酒中的萜烯类化合物定量分析。

调配型五味子药酒工艺研究

[目的]以五味子为主要原料、五味子乙素含量为指标,研制调配型五味子药酒。[方法]以正交试验优化五味子提取工艺,通过调配得到五味子药酒。[结果]五味子乙素最佳提取工艺为:乙醇浓度80%,料液比1∶8,提取时间3 h,提取3次,五味子乙素提取率达4.93%。五味子低、高度药酒的影响因素顺序均为柠檬酸添加量、蜂蜜用量、蔗糖用量。其中,低度五味子药酒的最佳配方为:按体积添加5.0%的蔗糖、0.2%的柠檬酸、0.2%的蜂蜜;高度五味子药酒的最佳配方为:按体积添加5.0%的蔗糖、0.2%的柠檬酸、0.4%的蜂蜜。[结论]按此最优工艺得到的五味子药酒甜酸适口、风味较好。

英平诸痹灵药酒中铜、锰、镍、钴、锶、锂的含量及分析

利用火焰原子吸收法测定 5种英平诸痹灵药酒中铜、锰、镍、钴、锶、锂的含量。结果表明 ,该类药酒中锰含量较丰富 ,锂含量较低 ,其含量由多到少的顺序为 :锰 >锶 >镍 >铜 >钴 >锂。探讨了英平诸痹灵药酒中微量元素与治疗类风湿疾病的关系。

人参蚂蚁药酒对小鼠的抗疲劳作用

目的:研究人参蚂蚁药酒对小鼠的抗疲劳作用。方法:将150只雄性昆明种小鼠按体重随机分为5组,分别灌胃不同剂量的人参蚂蚁药酒及红景天溶液(阳性对照组),同样体积蒸馏水(阴性对照组),14 d后称量小鼠体重,分别检测小鼠力竭游泳时间,运动后血清尿素氮、全血乳酸以及肝、肌糖原含量。结果:灌胃15 d后人参蚂蚁药酒对小鼠的体重增长无明显影响。经人参蚂蚁药酒处理的小鼠力竭游泳时间明显高于阴性对照组小鼠,而运动后血清尿素氮和血乳酸浓度明显低于阴性对照组,肝糖原和肌糖原均显著高于阴性对照组,与阳性对照组相似。结论:人参蚂蚁药酒对小鼠有明显的抗疲劳效果。

七味药酒治疗压疮的观察与护理体会

目的:观察七味药酒联合护理治疗压疮的临床疗效及护理体会。方法:112例压疮患者随机分为治疗组和对照组各56例,2组均采用压疮常规护理,在此基础上,治疗组加用七味药酒外敷,观察2组临床疗效和伤口愈合时间、压疮愈合计分表评分差异。结果:治疗组总有效率91.07%;对照组总有效率75%;2组比较,有统计学意义(P<0.05);组间治疗后比较,治疗组伤口愈合时间和PUSH评分较对照组相比明显缩短,2组比较,有统计学意义(P<0.05)。结论:七味药酒联合护理治疗压疮疗效确切,值得临床推广应用。

超滤法精制生脉饮口服液初探

通过生脉饮口服液的超滤澄清与精制，对采用传统工艺和超滤技术制备的成品制剂在澄清度、除杂降浊效果、有效成份含量等方面进行了质量考察，并对温度、料液流速等因素进行研究。结果表明，超滤技术不仅能除去杂质并能较为有效地保留原方配伍成份，可用于复方中药口服液的精制。

中药912液对急性肺损伤大鼠干预作用的实验研究

目的 :从细胞凋亡角度探讨急性肺损伤 (AL I)发病机制及中药 912液的干预作用。方法 :Wistar大鼠46只 ,随机分成 3组 :1模型组 (AL I组 ) 30只 ,静注标准大肠杆菌菌株制成 AL I模型 ,分别于给药后 4、8及 12小时采集样品 (各时间点 10只 ) ;2假手术组 (SO组 ) 8只 ,静注等量生理盐水 ,12小时采样 ;3中药组 (912液组 ) 8只 ,制模半小时后再静注 912液 ,12小时采样。以肺组织病理切片、血气氧分压 (Pa O2 )、肺系数 (L I)、肺通透指数 (L PI)及肿瘤坏死因子α(TNFα)监测模型 ;以电镜、流式细胞仪术和 TU NEL法检测细胞凋亡。结果 :AL I组给药后 4小时起光镜下可见肺组织肺泡腔狭窄、中性粒细胞渗出等 AL I表现 ,且随时程延长病变加重。 Pa O2 降低 ,L I、L PI及 TNFα增高 ,与 SO组相比有显著性差异 (P均 <0 .0 1)。凋亡结果 TUNEL 染色AL I组部分肺组织细胞呈阳性染色 ,流式细胞术显示 AL I组 4、8及 12小时凋亡细胞分别为 (7.15± 1.2 6 ) % ,(12 .39± 1.6 7) % ,(6 .72± 1.11) % ,呈先增加后下降趋势 ,与 SO组相比 ,P均 <0 .0 1。 912液组对肺功能各项指标均有明显改善 ,凋亡细胞数增高 (2 4.6 7± 2 .85 ) % ,与 AL I组相比 ,有显著性差异 (P<0 .0 1)。结论 :细胞凋亡在 AL I发生机制中起重要作用?

附子药酒抗肿瘤初步研究

目的探讨附子药酒的抗肿瘤作用。方法体内实验:建立小鼠肝癌H22皮下移植动物模型,灌胃给药后测定原位肿瘤体积、质量及免疫器官指数。用试剂盒测定血清中过氧化氢酶(CAT)、肌酸激酶(CK)、总超氧化物歧化酶(T-SOD)的活力,并通过基质金属明胶酶谱电泳考察附子药酒对基质金属蛋白酶的影响。体外实验:用MTT法测定附子药酒对小鼠乳腺癌4T1细胞和小鼠脾淋巴细胞增殖的影响。结果体内实验:在本实验剂量下,与模型组比较,附子组肿瘤生长减缓,抑瘤率为15.65%,胸腺指数、脾脏指数均降低;与模型组比较,附子药酒增强CAT和T-SOD活力,降低CK活力,并使基质金属酶-2(MMP-2)表达显著提高。体外实验:实验所用浓度的附子药酒对小鼠乳腺癌细胞无直接生长抑制作用,对小鼠脾淋巴细胞增殖无影响。结论附子药酒虽有一定抗肿瘤作用,也有对阻止肿瘤生长不利的方面,在肿瘤治疗中建议与其他中药联合使用。