Differential expression pattern, bioinformatics analysis, and validation of circRNA and mRNA in patients with arteriosclerosis.

Background: Lower limb arteriosclerosis obliterans (ASO) is the formation of atherosclerotic plaques in lower limb arteries, leading to vascular stenosis and occlusion, and is a major factor leading to lower limb amputation. The ASO seriously endangers the physical and mental health of patients. As living standards improve, the disease tends to occur in younger patients, and the incidence keeps increasing year by year. The circular RNAs (circRNAs) have been found to be tissue-specific, and they play an important role in a variety of diseases, but there are few studies on the pathogenic role and expression of circRNAs in ASOs. Method: Three diseased arteries from patients with ASO and three healthy arteries from healthy donors were collected for second-generation sequencing, and the pathogenic pathways and possible pathogenic circRNAs related to ASO were screened through bioinformatics analysis. PCR and agarose gel electrophoresis were used to validate the sequencing results. The expression of circRNA-0008706 in human arterial smooth muscle cells (HASMCs) was knocked down using siRNA technology to explore its function. Result: We identified 480 differentially expressed (DE) circRNAs and 2,997 DEmRNAs. Functional analysis revealed that epithelial-to-mesenchymal transition (EMT), lipid transport, regulation of extracellular matrix disassembly, regulation of cardiac muscle cell proliferation, branched-chain amino acid biosynthetic process, and positive regulation of cell growth and migration were enriched. Based on our previous microRNA array results, we constructed an ASO disease-specific competing endogenous (ceRNA) network. After validation, circRNA-0008706 was selected for functional analysis. Knockdown of circRNA-0008706 significantly suppressed the proliferation and migration phenotype of HASMCs and decreased the BCAT1 expression, which may be due to the specific binding of circRNA-0008706 to microRNA-125b-5p. Conclusion: This study is the first to compare the circRNA and mRNA expression profiles of ASOs and healthy arterial specimens and to construct a disease-specific ceRNA network for ASOs. This study may provide a new therapeutic target for ASO.

Combination of intramuscular transplantation of autologous mononuclear bone marrow cells with sympathectomy (L2, 3) in patients with peripheral arterial disease(PAD).

BACKGROUND: In most patients with severe, chronic extremity ischemic diseases, intervention or surgical treatment is often not suitable. Combination of intramuscular transplantation of autologous monocular bone marrow cells (AMBMCs) and sympathectomy (L2, 3) has been proved therapeutically beneficial. METHODS: We studied 170 patients (combined group 80, control group 90) with extremity ischemia (TAO, ASO FontaineⅡ,Ⅲ, Ⅳ) between January 2013 and September 2019. RESULTS: In contrast to pre-operation, the walking distance of patients increased significantly (from 61.34 ± 52.23 m to 156.0 ± 32.4 m, p < 0.01), and the ankle-brachial index (ABI) remarkably improved (from 0.28 ± 0.13 to 0.59 ± 0.23, p < 0.05). CONCLUSION: Combined therapy is feasible and effective for patients with peripheral arterial disease (PAD).

[Warming needling combined with Zhuyu Tongluo Xuebi decoction for arteriosclerosis obliterans with stasis].

OBJECTIVE: To observe the effect difference of warming needling combined with Zhuyu Tongluo Xuebi decoction and ciloprost for arteriosclerosis obliterans (ASO) with stasis. METHODS: A total of 96 ASO patients with stasis were randomly assigned into a combination group and a western medication group, 48 cases in each group. Anti-hypertension, glucose-lowering and lipid lowering therapies were applied in the two groups. Ciloprost was prescribed orally in the western medication group, twice a day, 100 mg a time. The main acupoints in the combination group were Sanyinjiao (SP 6), Yinlingquan (SP 9), Zusanli (ST 36), Guanyuan (CV 4), and Xuehai (SP 10), matched with Yanglingquan (GB 34) and Weizhong (BL 40). Warming needling was used at Sanyinjiao (SP 6), Zusanli (ST 36), Xuehai (SP 10) and Guanyuan (CV 4), 5 times a week, once a day, 20 min a time. At the same time, self-made Zhuyu Tongluo Xuebi decoction was applied in the combination group, 1 dose a day, twice a day. All the treatment was given for continuous 3 courses, 1 month as a course. The indexes were the symptom scores for cool limb skin, sour swelling, numbness, pain, abnormal complexion, ankle brachial index (ABI) and blood biochemical indexes, including fasting blood-glucose (FPG), triacylglycerol (TG), cholesterol total (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminase (AST), serum creatinine (Scr) and blood urea nitrogen (BUN). The adverse reactions were recorded. The clinical effect was evaluated. Two-month follow-up was carried out. RESULTS: After treatment, the symptom scores for cool limb skin, sour swelling, numbness, pain, abnormal complexion and total score decreased in the two groups (all P<0.05), with better results in the combination group (all P<0.05). The bilateral ABI were higher than those before treatment in the two groups (all P<0.05), with better results in the combination group (both P<0.05). The FPG, TG, TC, HDL-C, LDL-C, ALT, AST, Scr, BUN before and after treatment had no statistical significance in the two groups (all P>0.05). There was no adverse reaction on acupuncture and moxibustion. The total effective rate of the combination group was 95.8% (46/48), which was better than 91.7% (44/48) of the western medication group (P<0.05). The recurrence and aggravation rate in the combination group was 8.7% (4/46), which was lower than 18.2% (8/44) in the western medication group (P<0.05). CONCLUSION: Warming needling combined with Zhuyu Tongluo Xuebi decoction for ASO are better than simple oral ciloprost, with safety.

Warming needling combined with decoction for arteriosclerosis obliterans with stasis / 中国针灸

<p><b>OBJECTIVE</b>To observe the effect difference of warming needling combined with decoction and ciloprost for arteriosclerosis obliterans (ASO) with stasis.</p><p><b>METHODS</b>A total of 96 ASO patients with stasis were randomly assigned into a combination group and a western medication group, 48 cases in each group. Anti-hypertension, glucose-lowering and lipid lowering therapies were applied in the two groups. Ciloprost was prescribed orally in the western medication group, twice a day, 100 mg a time. The main acupoints in the combination group were Sanyinjiao (SP 6), Yinlingquan (SP 9), Zusanli (ST 36), Guanyuan (CV 4), and Xuehai (SP 10), matched with Yanglingquan (GB 34) and Weizhong (BL 40). Warming needling was used at Sanyinjiao (SP 6), Zusanli (ST 36), Xuehai (SP 10) and Guanyuan (CV 4), 5 times a week, once a day, 20 min a time. At the same time, self-made decoction was applied in the combination group, 1 dose a day, twice a day. All the treatment was given for continuous 3 courses, 1 month as a course. The indexes were the symptom scores for cool limb skin, sour swelling, numbness, pain, abnormal complexion, ankle brachial index (ABI) and blood biochemical indexes, including fasting blood-glucose (FPG), triacylglycerol (TG), cholesterol total (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminase (AST), serum creatinine (Scr) and blood urea nitrogen (BUN). The adverse reactions were recorded. The clinical effect was evaluated. Two-month follow-up was carried out.</p><p><b>RESULTS</b>After treatment, the symptom scores for cool limb skin, sour swelling, numbness, pain, abnormal complexion and total score decreased in the two groups (all <0.05), with better results in the combination group (all <0.05). The bilateral ABI were higher than those before treatment in the two groups (all <0.05), with better results in the combination group (both <0.05). The FPG, TG, TC, HDL-C, LDL-C, ALT, AST, Scr, BUN before and after treatment had no statistical significance in the two groups (all >0.05). There was no adverse reaction on acupuncture and moxibustion. The total effective rate of the combination group was 95.8% (46/48), which was better than 91.7% (44/48) of the western medication group (<0.05). The recurrence and aggravation rate in the combination group was 8.7% (4/46), which was lower than 18.2% (8/44) in the western medication group (<0.05).</p><p><b>CONCLUSION</b>Warming needling combined with decoction for ASO are better than simple oral ciloprost, with safety.</p>

Clinical features and predictors of patients with critical limb ischemia who responded to autologous mononuclear cell transplantation for therapeutic angiogenesis.

The clinical features of patients with critical limb ischemia (CLI) who responded to angiogenesis using autologous peripheral blood mononuclear cell transplantation (PB-MNC) have not yet been fully characterized, and there are no useful predictors to judge the curative effect in the early period after PB-MNC. This study sought to clarify the clinical features and predictors in patients with CLI who were successfully treated using PB-MNC. 30 consecutive patients [arteriosclerosis obliterans: 24 patients, thromboangiitis obliterans: 6 patients] who were diagnosed with major amputation despite maximal medical therapy were enrolled in this study. The study endpoint was major amputation within 3 months after PB-MNC. The collected data were evaluated for correlation between patients with and without major amputation within 3 months after PB-MNC. Six patients underwent major amputation and 1 patient underwent minor amputation. In the patients with major amputation, transcutaneous oxygen tension before PB-MNC and transplanted CD34-positive cells were lower than those of patients without major amputation. In the patients with amputation, interleukin-6 (IL-6) continued to increase after the first PB-MNC, and basic fibroblast growth factor (bFGF) decreased within 3 days after the first PB-MNC. PB-MNC was useful for the patients who were managed for inflammation and who had revascularization of the upper-popliteal arteries and two of the infra-popliteal arteries by endovascular and/or surgical revascularization. Variation in IL-6 and bFGF in the early period after PB-MNC could be useful predictors for the requirement of amputation within 3 months after PB-MNC.

Effect of Foot Baths containing Artificial CO2 on Peripheral Circulation / 日本温泉気候物理医学会雑誌

<b>Purpose</b>: We examined the effects of foot baths containing artificial carbon dioxide (CO₂) on peripheral circulation. Our goal was to obtain quantitative results bearing on the treatment and preservation of refractory leg ulcers and gangrene, which would occur in arteriosclerosis obliterans (ASO) and diabetic peripheral circulation disorders.<br><b>Procedures</b>: The lower legs of six healthy volunteers were placed for 10min into a 42°C bath of 10<i>l</i> in which 45g of artificial CO₂ “BUB” was dissolved. A laser Doppler blood-flowmeter was firmly attached. The measurements of blood flow were taken before, during, and 5, 15, 25, and 35min after each subject took a foot bath. Next, in order to examine the effects of peripheral blood flow on the whole body during the foot baths, each laser Doppler blood-flowmeters was attached on the subject's right arm, 10cm above the wrist joint.<br><b>Results</b>: The blood flow increased during the foot bath and decreased after the bath was removed; during in a 10min bath, the flow was 264±135 (%) (p<0.05) of the flow before the feet were placed into the bath. 5min after the bath was removed, the flow was 256±174 (%) of the original pre-bath rate. 15min later it was 146±60 (%), 25min later it was 112±23 (%), and 35min later it was 107±24 (%), as low as the flow before the feet were placed in the bath. The arm blood flow also increased during the foot bath and decreased after the bath was removed. During a 10min bath, the flow was 119±49 (%) of the rate before the feet were placed in the bath. 5min after the bath was removed, the flow was 120±66 (%) of the original pre-bath rate. 15min later it was 113±28 (%), 25min later it was 109±16 (%), and 35min later it was 95±14 (%), essentially the same as the flow before the feet were placed in the bath.<br><b>Discussion</b>: Our work demonstrates an increase in blood flow, by 2.6X, by using a foot bath containing artificial CO₂. The increase was maintained for 5min after removed of the foot from the bath. However, the blood flow decreased 15min after the baths were removed. We hope that this simple and effective foot bath will be used both at home and elderly care service stations. Its use could see prevention from refractory leg ulcers or gangrene, particularly as average life span continues to increase in Japan.