Coexistence of a nonresistance-conferring IncI1 plasmid favors persistence of the blaCTX-M-bearing IncFII plasmid in Escherichia coli.

The interaction between coexisting plasmids can affect plasmid-carried resistance gene persistence and spread. However, whether the persistence of the blaCTX-M gene in clinical Enterobacteriaceae is related to the interaction of coresident nonresistance-conferring plasmids has not been reported. This study was initiated to elucidate how a nonresistance-conferring IncI1 plasmid affected the blaCTX-M-bearing IncFII plasmid colocated on the same cell. Herein, we constructed three isogenic derivatives of E. coli C600, designated as C600FII, C600I1, and C600FII+I1, which harbored the blaCTX-M-IncFII plasmid and/or the nonresistance-IncI1 one. We discovered that strain C600FII+I1 conferred higher fitness advantages than strain C600FII; also, the stability of the blaCTX-M-IncFII plasmid was noticeably improved in an antibiotic-free environment when it coexisted with the IncI1 plasmid. To further explore why the IncI1 plasmid enhanced the persistence of the blaCTX-M-IncFII plasmid, we assessed the blaCTX-M-IncFII plasmid's copy numbers, conjugation frequencies, and rep gene expressions in strains C600FII and C600FII+I1. The results demonstrated that the rep expressions of the blaCTX-M-IncFII plasmid in strain C600FII+I1 was greatly decreased, along with the plasmid's copy numbers and mating efficiencies, compared to those in strain C600FII. Moreover, further study revealed that the intracellular ATP levels of strain C600FII+I1 were far lower than those of strain C600FII. Our findings confirmed that coexistence of the nonresistance-IncI1 plasmid can keep the blaCTX-M-IncFII plasmid more stable by increasing the fitness advantages of the host bacteria, which will pose a threat to preventing the long-term presence of the plasmid-carried blaCTX-M gene in clinical Enterobacteriaceae. IMPORTANCE: So far, plasmid-carried blaCTX-M is still the most common extended-spectrum beta-lactamase (ESBL) genotype in clinical settings worldwide. Except for the widespread use of third-generation cephalosporins, the interaction between coexisting plasmids can also affect the long-term stable existence of the blaCTX-M gene; however, the study on that is still sparse. In the present study, we assess the interaction of coinhabitant plasmids blaCTX-M-IncFII and nonresistance-IncI1. Our results confirmed that the increased fitness advantages of strain C600FII+I1 were attributable to the cohabitant nonresistance-IncI1 plasmid, which largely reduced the intracellular ATP levels of host bacteria, thus decreasing the rep gene expression of the blaCTX-M-IncFII plasmid, its copy numbers, and mating efficiencies, while the higher fitness advantages of strain C600FII+I1 enhanced the persistence of the blaCTX-M-IncFII plasmid. The results indicate that the nonresistance-IncI1 plasmid contributes to the long-term existence of the blaCTX-M-IncFII plasmid, implying a potentially new strategy for controlling the spread of resistance plasmids in clinical settings by targeting nonresistance plasmids.

Shengxian decoction improves lung function in rats with bleomycin-induced idiopathic pulmonary fibrosis through the inhibition of PANoptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Shengxian decoction (SXD) is a classic Chinese medicinal formula that can effectively improve clinical symptoms and quality of life and delay disease progression in idiopathic pulmonary fibrosis (IPF) patients; however, the underlying mechanisms remain unclear. AIM OF THE STUDY: This study aimed to observe PANoptosis in bleomycin-induced IPF and to assess the efficacy and mechanism of action of SXD in the treatment of IPF. MATERIALS AND METHODS: Fifty SD rats were randomly divided into the sham, IPF, IPF + pirfenidone (PFD), IPF + SXD-medium dose (SXD-M), and IPF + SXD-low dose (SXD-L) groups. Lung function analysis and microcomputed tomography imaging of the rats with IPF treated with oral pirfenidone or oral SXD for 28 days were performed. Hematoxylin and eosin (HE) staining and Masson's trichrome staining were used to observe pathological lung damage. Enzyme-linked immunosorbent assays (ELISAs) were used to determine the serum levels of IL-1ß, IL-18, TNF-α, and IFN-γ. Pyroptosis, apoptosis, and necroptosis were assessed using TUNEL, TUNEL/caspase-1, and PI fluorescence staining, respectively. GSDMD, caspase-3, and MLKL were examined by immunohistochemistry. The expression of fibrin-, ZBP1-, pyroptosis-, apoptosis-, and necroptosis-related proteins in the lung tissue was determined by western blotting. RESULTS: SXD normalized lung function in rats with bleomycin-induced IPF and reduced serum inflammatory factor levels and lung tissue fibrosis. The underlying mechanism of action involves the inhibition of pyroptosis pathway proteins, such as NLRP3, caspase-1, cleaved caspase-1, and GSDMD; apoptotic pathway proteins, such as Bax, Bcl-2, cleaved caspase-3, and caspase-3; and necroptosis pathway proteins, such as RIPK1, RIPK3, p-MLKL and MLKL. These pathways are modulated by the PANoptosis initiator ZBP1. Notably, the efficacy of SXD is concentration dependent, with a medium dose exhibiting superior effectiveness compared to a low dose. CONCLUSION: Bleomycin induced PANoptosis in the lung tissue of rats with IPF. Additionally, SXD effectively delayed or reversed the early pathological changes in bleomycin-induced pulmonary fibrosis by inhibiting PANoptosis.

Characterization of a Tigecycline-Resistant and blaCTX-M-Bearing Klebsiella pneumoniae Strain from a Peacock in a Chinese Zoo.

In Chinese zoos, there are usually specially designed bird parks, similar to petting zoos, that allow children and adults to interact with diverse birds. However, such behaviors present a risk for the transmission of zoonotic pathogens. Recently, we isolated eight strains of Klebsiella pneumoniae and identified two blaCTX-M-positive strains from 110 birds, including parrots, peacocks, and ostriches, using anal or nasal swabs in a bird park of a zoo in China. There, K. pneumoniae LYS105A was obtained from a diseased peacock with chronic respiratory diseases by a nasal swab, which harbored the blaCTX-M-3 gene and exhibited resistance to amoxicillin, cefotaxime, gentamicin, oxytetracycline, doxycycline, tigecycline, florfenicol, and enrofloxacin. According to an analysis by whole-genome sequencing, K. pneumoniae LYS105A belongs to serotype ST859 (sequence type 859)-K19 (capsular serotype 19) and contains two plasmids, of which pLYS105A-2 can be transferred by electrotransformation and harbors numerous resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91. The above-mentioned genes are located in a novel mobile composite transposon, Tn7131, which makes horizontal transfer more flexible. Although no known genes were identified in the chromosome, a significant increase in SoxS upregulated the expression levels of phoPQ, acrEF-tolC, and oqxAB, which contributed to strain LYS105A acquiring resistance to tigecycline (MIC = 4 mg/L) and intermediate resistance to colistin (MIC = 2 mg/L). Altogether, our findings show that bird parks in zoos may act as important vehicles for the spread of multidrug-resistant bacteria from birds to humans and vice versa. IMPORTANCE A multidrug-resistant ST859-K19 K. pneumoniae strain, LYS105A, was obtained from a diseased peacock in a Chinese zoo. In addition, multiple resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91 were located in a novel composite transposon, Tn7131, of a mobile plasmid, implying that most of the resistance genes in strain LYS105A can be moved easily via horizontal gene transfer. Meanwhile, an increase in SoxS can further positively regulate the expression of phoPQ, acrEF-tolC, and oqxAB, which is the key factor for strain LYS105A to develop resistance to tigecycline and colistin. Taken together, these findings enrich our understanding of the horizontal cross-species spread of drug resistance genes, which will help us curb the development of bacterial resistance.

Decreased cyclic-AMP caused by ATP contributes to fosfomycin heteroresistance in avian Escherichia coli.

BACKGROUND: Fosfomycin is an important broad-spectrum bactericidal antibiotic to treat multidrug-resistant bacteria infections. It is generally accepted that heteroresistant bacteria are an intermediate stage in the formation of drug resistance, but there are few studies on the formation mechanism underlying fosfomycin heteroresistance (FHR). OBJECTIVES: To reveal the characteristics and formation mechanisms of FHR in Escherichia coli isolates obtained from chickens. METHODS: We identified the FHR according to the population analysis profile (PAP) test and in vitro time-kill assay. Growth curves for FHR E. coli and their subpopulations were measured. Also, the subpopulations were repeatedly cultured in fosfomycin-free medium for 5-20 overnight incubation periods. The formation mechanisms of FHR in E. coli isolates were identified through accumulation assay, carbohydrate utilization testing, real-time relative quantitative PCR analysis, DNA sequencing, transcriptomic analysis, intracellular ATP and cAMP-level assessment. RESULTS: Four of six E. coli strains were confirmed to show FHR, with a total of six subpopulations. The subpopulations restored phenotypic susceptibilities to fosfomycin within 5-20 overnight incubation sessions, but four of six subpopulations still maintained FHR characteristics. Differing from their parental isolates, the uptake of fosfomycin in the subpopulations through GlpT was reduced remarkably. Further studies identified that the low expression of glpT was due to the decrease of intracellular cAMP levels in the subpopulations, which was caused by the decreased ATP levels in cells. CONCLUSIONS: Our findings revealed the formation mechanism of E. coli isolates showing FHR obtained from chicken in China and characterized the dynamic change traits in vitro of the subpopulations.

Thoughts on the effects of moxa smoke in the epidemic prevention.

Through summarizing and analyzing the modern mechanism researches and controversial questions of moxibustion fumigation in the epidemic prevention, the thoughts on tackling the critical points are proposed in the paper, such as the recognition of moxa smoke in traditional Chinese medicine (TCM), the mechanism of moxa smoke in air disinfection and the characteristics of clinical application of moxa smoke so as to provide the references to the prevention and control of COVID-19 in TCM.

[Control observation between mild moxibustion and TDP for obsolete collateral ligament injury of interphalangeal joints].

OBJECTIVE: To compare the effects between mild moxibustion and specific electromagnetic spectrum therapy apparatus (TDP) for obsolete collateral ligament injury of interphalangeal joints. METHODS: Sixty patients were randomly divided into a mild moxibustion group and a TDP group, 30 cases in each one. In the mild moxibustion group, pure moxa sticks were used at the affected digital joints locally for 20-30 min a time. In the TDP group, TDP was applied at the affected digital joints locally for 20-30 min a time. The treatment was given once a day for two courses, and 10-day treatment was made into a course. Visual analogue scale (VAS) for pain, swelling degree of the affected digital joints before and after treatment were observed and the clinical efficacy and safety were evaluated in the two groups. RESULTS: The excellent rate was 56.7% (17/30) and the excellent and, good rate was 83.4% (25/30) in the mild moxibustion group,which were better than 36.7% (11/30) and 76.7% (23/30) in the TDP group respectively (both P < 0.01). After treatment the score of VAS and digital joints swelling degree were improved than those before treatment in the two groups (P < 0.01, P < 0.05), and the improvements of the mild moxibustion group were better than those of the TDP group (P < 0.01, P < 0.05). CONCLUSION: Mild moxibustion can apparently relieve the painful and swelling degree of obsolete collateral ligament injury of interphalangeal joints, which is superior to TDP.

[Influence of Electroacupuncture Intervention on Glutamic Acid and Ca2+ Contents and Expression of NMDA Receptor Protein in Hippocampus in Vascular Dementia Rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) intervention on hippocampal glutamate (Glu) and Ca2+ contents, and expression of Glu-N-methyl-D-aspartic acid receptor(NMDAR) and the learning-memory ability in vascular dementia (VD) rats, so as to reveal its mechanisms underlying improvement of VD. METHODS: SD rats were rando-mized into sham operation (sham) group (n=9), model group (n=11) and EA groups (n=10). The VD model was established by repeated bilateral common carotid arteries occlusion and reperfusion plus intraperitoneal injection of sodium nitroprusside. EA (2 Hz, 2 mA) was applied to "Baihui" (GV 20)-"Housanli" (ST 36) and "Geshu" (BL 17)-"Dazhui" (GV 14) for 10 min, once a day for 15 consecutive days. The neurological function was assessed by using stroke index (0-10 points) and neurological deficit scaling(0-10 points). The learning-memory ability was evaluated by using step-down tests. The contents of Glu and Ca2+ in the right hippocampal tissue were determined by using aspectrophotometer and the expression of NMDAR protein in the right hippocampus was detected by immunoblotting. RESULTS: Compared with the sham group, the stroke index and neurological deficit scores, and the reaction latency and the error times of step-down tests, as well as the contents of Glu and Ca2+ and the expression level of NMDAR in the right hippocampus were significantly increased in the model group (P<0.05, P<0.01), while the step-through latency was considerably decreased (P<0.01), suggesting a neurological disorder and a cognitive decline. After EA intervention, the reaction latency and error times of step-down tests, the contents of Glu and Ca2+ and the expression level of NMDAR in the right hippocampus were significantly down-regulated, and the step-through latency was notably increased in comparison with the model group (P<0.01). CONCLUSIONS: EA intervention is able to improve the cognitive ability of VD rats, which may be associated with its effects in reducing the excitatory neurotoxicity of hippocampal Glu-NMDAR and lowering cellular Ca2+ load to resist neuronal injury.

[Body acupuncture combined with auricular acupressure for menstrual headache: a randomized controlled clinical trial].

OBJECTIVE: To observe the therapeutic effect of body acupuncture combined with auricular acupressure for treatment of menstrual headache of hyperactivity of "liver fire". METHODS: A total of 85 menstrual headache patients with hyperactivity of "liver-fire" were randomly divided into control group (n = 42) and treatment group (n = 43). Patients of the control group were treated by oral administration of Flunarizine Hydrochloride Capsules (Sibelium, 5 mg/time, twice daily) for 5 days beginning at the headache attack in the first menstrual cycle, and for two weeks (except weekends) in the second and third menstrual cycles. Patients of the treatment group were treated by manual acupuncture stimulation of body acupoints Baihui (GV 20), Fengchi (GB 20), Sanyinjiao (SP 6), Taichong (LR 3), Xiaxi (GB 43), Taiyang (EX-HN 5), Hanyan (GB 4), Xuanlu (GB 5) and Shuaigu (GB 8) beginning from the headache attack, and otopoint-pellet pressure of otopoints unilateral Endocrine, Ovaries, Shenmen, Cortex, Liver, Spleen and Kidney (beginning 5 days before menstrual onset) and bilateral Shenmen, Nie, Liver, Gallbladder, Subcortex and Jiaogan during headache attack. Clinical symptom scores were assessed according to the "Guiding Principles for Clinical Research on New Chinese Herbal Drugs" issued by the Chinese Ministry of Health. RESULTS: After the treatment, of the 42 and 43 cases in the control and treatment groups, 9 (21.43%) and 20 (46.51%) were cured, 12 (28.57%) and 14 (32.56%) had a marked improvement, 13 (30.95%) and 7 (16.28%) were effective, 8 (19.05%) and 2 (4.65%) invalid, with the effective rates being 80.95% and 95.35% , respectively. The curative effect of the treatment group was significantly better than that of the control group (P<0.05). The symptom score and VAS score after the treatment and 3 months post-treatment in the treatment group were significantly lower than those of the control group (P<0.01). The headache scores and VAS scores were significantly reduced following the treatment and 3 months after the treatment in both control and treatment groups (P<0.01). After the treatment for 2-3 therapeutic courses, the headache attack duration during menstruation was markedly and gradually shortened in both groups (P<0.01), and therapeutic effect of the treatment group was significantly better than that of the control group (P<0.01). CONCLUSION: Body acupuncture combined with auricular acupressure treatment can effectively relieve menstrual headache in menstrual women with hyperactivity of "liver fire".

[Efficacy of acupuncture combined with auricular point sticking on the content of serum prostaglandin F2α, and plasma arginine vasopressin in patients with menstrual headache].

OBJECTIVE: To observe the clinical efficacy of acupuncture combined with auricular point sticking for menstrual headache and to discuss its mechanism. METHODS: Eighty-five patients with menstrual headache were randomly divided into an observation group (43 cases) and a control group (42 cases). The observation group was treated with body acupuncture combined with auricular point sticking and the control group was treated with flunarizine hydrochloride capsules orally. The treatments of 3 menstrual cycles were required. The clinical efficacy was observed in the two groups. The content of serum prostaglandin F2α, (PGF2α) and plasma arginine vasopressin (AVP) in the menstrual periods of some patients randomly selected in the two groups was tested before and after treatment and was compared with that of 20 cases in a normal group. Results The total effective rate was 95.4% (41/43) in the observation group which was obviously superior to 81.0% (34/42) in the control group (P<0.01). Before treatment, the content of serum PGF2α and plasma AVP of patients in the two groups was higher than that in the normal group (all P<0.01). After treatment,the content of serum PGF2α and plasma AVP was lower than that before treatment in the two groups (P<0.01, P<0.05). The content of serum PGF2α in the observation group was decreased significantly compared with that in the control group (P<0.05) and returned to the level of the normal group. CONCLUSION: Body acupuncture combined with auricular point sticking achieves positive efficacy for menstrual headache and its mechanism could be related to regulating the abnormal levels of serum PGF2α and plasma AVP.

[Effect of electroacupuncture intervention at different time-points of post-modeling on hippocampal monoamine neurotransmitter levels in mice with vascular dementia].

OBJECTIVE: To observe the effect of electrocupuncture (EA) intervention at different time-points of post-modeling on behavior and hippocampal monoamine neurotransmitter noradrenalin (NE), dopamine (DA) and 5-hydroxytrypamine (5-HT) contents in vascular dementia (VD) mice, so as to study its mechanism underlying improvement of VD. METHODS: A total of 60 Kunming mice were randomized into sham-operation control (n = 20), VD model (n = 20), EA-day (D)-1 (EA treatment was given from the 1st day on after modeling, n = 10), EA-D-3 (EA was given from the 3rd day on after modeling, n = 10) groups. VD model was established by occlusion of the bilateral cervical common arteries and reperfusion. EA (2 Hz/80 Hz) was applied to "Baihui" (GV 20), "Dazhui" (GV 14),"Zusanli" (ST 36) and "Geshu"(BL 17) for 10 min, once daily for 15 days. Hippocampal NE, DA and 5-HT contents were assayed by fluorospectrophotometry. The mouse's learning-memory ability was assessed by step-down tests. RESULTS: In comparison with the sham-operation control group, the learning-memory ability (marked increase of reaction time and error times, decrease of step-down latency) was apparently lowered in the model group (P < 0.01). The hippocampal NE, DA and 5-HT contents were significantly lower in the model group than in the sham-operation group (P < 0.01). Compared with the model group, the mice's learning-memory ability (marked decrease of reaction time and error times, increase of step-down latency) was significantly increased in EA intervention groups (P < 0.01), and hippocampal NE, DA and 5-HT levels were significantly increased (P < 0.01), and the effect of EA-D-3 group was obviously better than that of the EA-D-1 group (P < 0.05, P < 0.01). CONCLUSION: EA can improve the VD mice's learning-memory ability, which is closely related to its effects in up-regulating hippocampal NE, DA and 5-HT contents, and the effect of later EA intervention after modeling is better.

[Effect of mild moxibustion on macrophage number and collagen expression of raw surface tissue in chronic refractory skin ulcer rats].

OBJECTIVE: To observe the effect of mild moxibustion on the number of macrophages and contents of collagen I and III in the raw surface tissue in chronic skin ulcer rats so as to study its mechanism underlying promoting scar formation. METHODS: Eighty male SD rats were randomly divided into normal (n = 8), model (n = 24), TDP (n = 24) and moxibustion (n = 24) groups. Chronic refractory skin ulcer was established by making an open wound at the back and local intramuscular injection of hydrocortisone sodium succinate. For rats of the TDP and moxibustion groups, TDP irradiation or mild moxibustion was applied to the raw surface, bilateral "Shenshu" (BL 23) and "Zusanli" (ST 36) for 15 min, once daily for 7, 10 and 14 days, respectively. The number of macrophages in the raw surface tissue was counted under light microscope after H. E. staining and col- lagen I and III expressions of the raw surface tissue were detected by immunohistochemistry. RESULTS: In comparison with the normal group, the numbers of macrophages in the raw surface tissue were increased significantly in the model group on day 7, 10 and 14 (P < 0.05); while compared with the model group, the numbers of macrophages were increased further obviously in the moxibustion group on day 7 and 10 and in the TDP group on day 10 after the treatment (P < 0.05). Compared with the model group, the numbers of macrophages in both TDP and moxibustion groups were down-regulated obviously (P < 0. 05). In regard to collagen I and III expression of the raw surface tissue, compared with the normal group, the collagen I protein expression level was down-regulated markedly in the model group on the 7th day (P < 0.01); whereas in comparison with the model group, the expression levels of collagen I and III were increased considerably in the TDP and moxibustion groups on day 7 and 14 after the treatment (P < 0.05, P < 0.01). The ratios of collagen I/III expression were remarkably higher in the model group than in the normal group on day 7 and 14 (P < 0.05), and significantly lower in the TDP group on day 7 and 14 and in the moxibustion group on day 14 than in the model group (P < 0.05, P < 0.01). The effects of moxibustion were obviously superior to those of TDP in up-regulating macrophage number on day 10, up-regulating collagen I and III expressions on day 14, and down-regulating macrophage number on day 14 after the treatment (P < 0.05, P < 0.01). No significant differences were found between the TDP and moxibustion groups in up-regulating macrophage number, and collagen I and III protein expressions, and in down-regulating the ratios of collagen I/III expression on day 7 after the treatment (P > 0.05). CONCLUSION: Mild moxibustion can regulate the number of macrophages and strengthen the expression of collagen proteins in the raw surface tissue in the chronic skin ulcer rats, which may contribute to its effect in promoting wound healing and reducing scar formation.

[Effect of electroacupuncture on lipid metabolism in male and female obesity rats].

OBJECTIVE: To investigate the influence of electroacupuncture (EA) on lipid metabolism in both sexes of obesity model rats. METHODS: A total of 30 male and 30 female SD rats were randomized into normal control, model and EA groups respectively, with 10 rats in each group. EA (2 Hz, 1-2 mA) was applied to the ipsilateral "Zusanli" (ST 36)-"Sanyin-jiao"(SP 6) and bilateral "Fenglong" (ST 40) for 30 min, once daily for 28 days. In addition,"Quchi" (LI 11),"Zhongwan" (CV 12) and "Guanyuan" (CV 4) were punctured with filiform needles and stimulated by manipulating the needle for 1 min, separately. Obesity model was duplicated by subcutaneous injection of 15% glutamic sodium (0.2 mL/10 g) once daily for 5 days and by feeding the animal with high fat forage for 36 days. Lee's index was calculated, and plasma triglyeride (TG) concentration detected by glycerol-phosphoric acid oxidase-peroxydase method, total cholesterol (TC) detected by cholesterol oxidase method, and plasma low-density lipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol (HDL-C) contents were assayed by one-step method, respectively. RESULTS: In both male and female rats, compared to their own normal groups, the Lee's index, plasma TG, TC and LDL-C contents were increased significantly and plasma HDL-C contents decreased obviously in the model groups (P<0.01), while compared to their own model groups, the Lee's index, plasma TG, TC and LDL-C levels in the EA groups were down-regulated considerably and plasma HDL-C contents up-regulated remarkably after the treatment (P<0.01, P<0.05). The efficacies of the EA intervention were obviously better in reducing Lee's index for male rats, and in down-regulating plasma TG and TC contents in female rats (P<0.05). No significant differences were found between male and female rats in EA intervention for plasma LDL-C and HDL-C levels. CONCLUSION: EA intervention can effectively regulate obesity rats' Lee's index and plasma TG, TC, LDL-C and HDL-C levels. The effects are different in male and female rats.

[Effect of mild-warm moxibustion on microcirculation in the raw surface tissue of chronic refractory wound in skin ulcer rats].

OBJECTIVE: To observe the effects of mild-warm moxibustion on dynamic blood flow, microvessel count (MVC)and vascular endothelial growth factor (VEGF) expression in the wound tissue of the chronic skin ulcer in rats, so as to reveal its underlying mechanism in promoting wound recovery. METHODS: A total of 104 male SD rats with skin injury were randomly divided into control group (n=8), model group (n=32), TDP (far-infrared heating device) group (n=32) and moxibustion group (n=32). Chronic refractory raw surface wound model was established by muscular injection of Hydrocortisone Sodium Succinate. For rats of the TDP and moxibustion groups, TDP irridiation and mild-warm moxibustion were applied to the raw surface, bilateral "Shenshu" (BL 23) and "Zusanli" (ST 36) for 15 min, once daily for 3, 7 and 14 days respectively. The healing rate and the healing time of raw surface of the wound were observed. The blood flow of the raw surface of the wound tissue was measured by laser Doppler flowmeter and the MVC in granulation tissue of chronic skin ulcer was counted under light microscope. VEGF expression was detected by immunohistochemistry. RESULTS: In comparison with the control group, the healing rate of the wound raw surface was significantly lower and the healing time was prolonged in the model group (P < 0.01). Compared with the model group, the healing rates on day 3, 7, 10 and 14 were significantly higher and the healing time was strikingly faster in both TDP and moxibustion groups (P < 0.01, P < 0.05), and the effects of the moxibustion group in increasing the healing rate and shortening the healing time were significantly better than those of TDP group (P < 0.01). In comparison with the model group, the blood flow volume, MVC and VEGF expression levels on day 3 and 7 were upregulated significantly in both TDP and moxibustion groups (P < 0.01, P < 0.05); while the blood flow volume, MVC and VEGF expression level in the moxibustion group and the blood flow volume and VEGF expression level in the TDP group downregulated considerably on day 14 (P < 0.01). No significant difference was found between the TDP and moxibustion groups in the MVC on day 14 after the treatment (P > 0.05). CONCLUSION: Mild-warm moxibustion can promote wound healing, which is closely with its effects in increasing blood flow and MVC, and upregulating VEGF expression in the wound granulation tissue of the chronic skin ulcer.