[Effect of wheat-grain moxibustion on the expression of Beclin-1/GRP78 in spinal dorsal horn in rats with cervical spondylotic radiculopathy].

OBJECTIVE: To observe the effect of wheat-grain moxibustion at "Dazhui" (GV 14) on the expressions of Beclin-1 and GRP78 in spinal dorsal horn in rats with cervical spondylotic radiculopathy (CSR), and to explore the possible analgesic mechanism of wheat-grain moxibustion for CSR. METHODS: A total of 48 SD rats were randomly divided into a sham operation group, a model group, a wheat-grain moxibustion group and a wheat-grain moxibustion+3-MA group, 12 rats in each group. The CSR model was prepared by spinal cord insertion method. Three days after modeling, the rats in the model group were intraperitoneally injected with 1 mL of 0.9% sodium chloride solution; the rats in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14, 6 cones per time) on the basis of the model group; the rats in the wheat-grain moxibustion+3-MA group were intraperitoneally injected with 3-MA solution and wheat-grain moxibustion at "Dazhui" (GV 14, 6 cones per time). The three groups were intervened for 7 days, once a day. The gait score and mechanical pain threshold were observed before treatment and 7 days into treatment; after the treatment, the expressions of mRNA and protein of Beclin-1 in spinal dorsal horn were detected by real-time fluorescence quantitative PCR and immunohistochemistry; the expression of GRP78 protein in spinal dorsal horn was detected by Western blot method; the autophagosomes and ultrastructure in spinal dorsal horn neurons were observed by electron microscope. RESULTS: After the treatment, compared with the sham operation group, in the model group, the gait score was increased and the mechanical pain threshold was decreased (P<0.01), and the expression of GRP78 protein in spinal dorsal horn was increased (P<0.01). Compared with the model group and the wheat-grain moxibustion+3-MA group, in the wheat-grain moxibustion group, the gait score was decreased and mechanical pain threshold was increased (P<0.01), and the expression of GRP78 protein in spinal dorsal horn was decreased, and the expressions of mRNA and protein of Beclin-1 were increased (P<0.01). Under electron microscope, the ultrastructure of spinal dorsal horn neurons in the wheat-grain moxibustion group was not significantly damaged, and its structure was basically close to normal, and the number of autophagosomes was more than the other three groups. CONCLUSION: Wheat-grain moxibustion at "Dazhui" (GV 14) has analgesic effect on CSR rats. The mechanism may be related to moderately up-regulate the expression of Beclin-1, enhance autophagy and reduce endoplasmic reticulum stress.

Direct moxibustion exerts an analgesic effect on cervical spondylotic radiculopathy by increasing autophagy via the Act A/Smads signaling pathway.

BACKGROUND: Direct moxibustion (DM) is reported to be useful for cervical spondylotic radiculopathy (CSR), but the analgesic mechanism remains unknown. Autophagy plays a protective role in neuronal apoptosis, Act A/Smads signaling pathway has been confirmed to be associated with the activation of autophagy. The study aimed to explore the effect of DM on autophagy in rats with CSR and the involvement of Act A/Smads signaling pathway. METHODS: Rats were randomly divided into Sham, CSR, CSR + DM, CSR + DM + 3-MA (PI3K inhibitor), and CSR + DM + SB (Act A inhibitor) group. Three days after establishment of CSR model with a fish line inserted under the axilla of the nerve roots, DM at Dazhui (GV14) was performed six times once for seven consecutive days. Western blot and immunofluorescence staining were used to observe the expression of the neuronal autophagy molecule LC3II/I, Atg7, and Act A/Smads signaling molecule Act A, p-Smad2, and p-Smad3. Bcl-2/Bax mRNA expression was measured by real time PCR. RESULTS: DM improved the pain threshold and motor function of CSR rats and promoted the expression of Act A, p-Smad2, p-Smad3, LC3II/I, and Atg7 in the entrapped-nerve root spinal dorsal horn. DM reduced the expression of Bax mRNA and decreased the number of apoptotic neurons. 3-MA and Act A inhibitor SB suppressed the expression of above-mentioned proteins and reduced the protective effect of DM on apoptotic neurons. CONCLUSION: DM exerts analgesic effects by regulating the autophagy to reduce cell apoptosis and repair nerve injury, and this feature may be related to the Act A/Smads signaling pathway.

[Effect of mild moxibustion on pain and expression of LC3 and Bax in spinal cord in cervical spondylotic radiculopathy rats].

OBJECTIVE: To observe the effect of mild moxibustion (Moxi) at "Dazhui" (GV14) on neuropathic pain, expression of autophagy and apoptosis factor LC3 and Bax proteins and mRNAs in the spinal cord tissue in rats with cervical spondylotic radiculopathy (CSR), so as to explore its underlying mechanism underlying relief of CSR-induced pain. METHODS: Forty rats (half male half female) were randomly divided into blank control, model, Moxi, Moxi+autophagy inhibitor 3-methyladenine (3-MA, Moxi+3-MA) groups, with 10 rats in each group. The CSR model was established by loose ligature of the local cervical nerve roots. Three days after modeling, mild Moxi was applied to GV14 for 10 min, once daily for 7 days. Rats of the Moxi+3-MA group received intraperitoneal injection of 3-MA(1 mL, 15 mg/kg+ saline) before Moxi, once daily for 7 consecutive days. Rats of the model and Moxi groups were also given normal saline (i.p., 1 mL), once daily for 7 days. The gait behavior score (1-3 points) was scaled according to the rats' pain reaction and foot paw contracture produced walking disorder and the mechanical pain threshold (MPT) was detected before and after the treatment. The expression of spinal cord LC3 and Bax proteins and mRNAs were detected by immunohistochemistry and quantitative RT-PCR, respectively. RESULTS: Compared with the blank control group, the gait disorder score, and percentage of Bax positive cells and expression of Bax mRNA were significantly increased (P<0.01, P<0.05), and MPT was markedly decreased in the model group (P<0.01). After the treatment, the gait disorder score, percentage of Bax positive cells and Bax mRNA expression were significantly down-regulated (P<0.01, P<0.05), while the MPT and percentage of LC3 positive cells and LC3 mRNA expression were considerably increased (P<0.01, P<0.05) in both Moxi and Moxi+3-MA groups. The therapeutic effects of mild Moxi were remarkably superior to those of Moxi+3-MA in downregulating gait disorder score, Bax positive cell percentage and Bax mRNA expression, and in up-regulating MPT, LC3 positive cell percentage and LC3 mRNA expression (P<0.05), suggesting a reduction of the function of mild Moxi after administration of 3-MA. CONCLUSION: Mild Moxi at GV14 can relieve neuropathic pain in CSR rats, which may be related to its functions in up-regulating LC3 autophagy, thereby inhibiting the expression of Bax pro-apoptotic protein in spinal cord to reduce apoptosis and to repair nerve injury.

Serotype distribution and drug resistance of Streptococcus pneumoniae in hospitalized patients in Yulin, Guangxi / 中国热带医学

@#Abstract: Objective　To analyze the serotype distribution, drug resistance rate and drug resistance gene carrying of Streptococcus pneumoniae isolates in hospitalized patients, and evaluate the coverage of the vaccine to the serotype of Streptococcus pneumoniae in this area, so as to provide reference for the rational use of antibiotics in clinic. Methods　A total of 150 strains of non-repetitive Streptococcus pneumoniae isolated from inpatients from January 2015 to December 2019 were collected for serotyping and antimicrobial sensitivity test. The carrying rates of pbp2b, ermB and tetM were detected by PCR. Results　The PCR classification rate of 150 strains of Streptococcus pneumoniae was 93.1%, and the classification rate of capsular swelling test was 100%, and a total of 19 serotypes were divided, mainly 19F and 6B. Children's serotypes were predominantly 19F, 6B, and 15A; adult serotypes were predominantly 19F, 14, and 23F. The coverage rates of the PCV7, PCV10, PCV13 and PPV23 vaccines were 36.8%, 42.1%, 57.9% and 68.4%, respectively. Strains with serotypes of 19F, 6B, 3, and 23F had higher rates of resistance to antimicrobials. The sensitivity of Streptococcus pneumoniae to penicillin was greater than 96.0%. Antimicrobials with significant differences in resistance rates between invasive and non-invasive strains were penicillin, moxifloxacin, and levofloxacin. The percentage of strains carrying both ermB and tetM resistance genes was 96.0%, and the concordance rate between pbp2b, ermB and tetM resistance genes and the resistance phenotype was >98.0%. A total of 10 multi-resistance combinations were detected, with a multi-resistance rate of 62.6%, and the multi-drug resistance pattern of Streptococcus pneumoniae was mainly concentrated in the 19F and 6B serotypes. Conclusion　There are significant age differences in the serotypes of Streptococcus pneumoniae in this area. The vaccine currently used has low coverage in this region and therefore offer limited protection to the population. The drug resistance rates of Streptococcus pneumoniae varied significantly among serotypes. Erythromycin and tetracycline are not recommended for clinical treatment of Streptococcus pneumoniae. Penicillin can still be used as the first choice for clinical treatment of Streptococcus pneumoniae infection.

[Therapeutic effect of acupuncture combined with wheat-grain moxibustion on perimenopausal depression with kidney deficiency and liver depression].

OBJECTIVE: To compare the therapeutic effect of acupuncture combined with wheat-grain moxibustion and oral fluoxetine hydrochloride capsule on perimenopausal depression with kidney deficiency and liver depression. METHODS: A total of 60 patients of perimenopausal depression with kidney deficiency and liver depression were randomized into an observation group and a control group, 30 cases in each one. Acupuncture combined with wheat-grain moxibustion were adopted in the observation group. Acupuncture was applied at Baihui (GV 20), Yintang (GV 29), Fengchi (GB 20), etc. for 30 min. Wheat-grain moxibustion was applied at Ganshu (BL 18), Shenshu (BL 23), Mingmen (GV 4) and Yongquan (KI 1), 3 moxa-cones for each acupoint. The treatment in the observation group was given once every 2 days, 3 times a week. Fluoxetine hydrochloride capsule was given orally in the control group, 20 mg each time, once a day. Treatment for 8 weeks was required in the both groups. The scores of Hamilton depression scale (HAMD) and modified Kupperman scale were observed in the both groups before and after treatment, and at 1, 3, 6-month follow-up. The therapeutic efficacy was evaluated after treatment. RESULTS: Compared before treatment, the scores of HAMD and modified Kupperman scale after treatment and at each time point of follow-up were decreased in the both groups (P<0.01), and the HAMD scores in the observation group were lower than the control group (P<0.01). The total effective rate was 93.3% (28/30) in the observation group, which was superior to 80.0% (24/30) in the control group (P<0.01). CONCLUSION: Acupuncture combined with wheat-grain moxibustion can effectively treat perimenopausal depression with kidney deficiency and liver depression, and have more stable and sustained therapeutic effect compared with oral fluoxetine hydrochloride capsule.

[Effects of mild moxibustion on Beclin-1/Bcl-2 expression in spinal cord of rats with cervical spondylotic radiculopathy].

OBJECTIVE: To observe the effects of mild moxibustion on the expression of autophagy and apoptosis factors Beclin-1, Bcl-2 mRNA and protein in spinal cord (including nerve root tissues) of cervical spondylotic radiculopathy (CSR) rats, so as to explore the analgesic mechanism of mild moxibustion at "Dazhui" (GV14) on CSR. METHODS: SD rats were randomly divided into blank group, model group, mild moxibustion group and mild moxibustion+3-methyladenine(3-MA) group, with 10 rats in each group. CSR model was established by inserting the wire into the cervical nerve root. The rats in the blank group were only fed normally without any intervention.The rats in the mild moxibustion group and mild moxibustion+3-MA group were given mild moxibustion at GV14 for 10 min each time,and intraperitoneal injection of 1 mL 0.9% normal saline and 1 mL 3-MA(15 mg/kg)separately. Rats in the model group were given 0.9% normal saline every day. All the three interventions were started from the 3rd day after modeling for 7 days. The rat's behavioral reaction of gait was scored and the pain threshold of rat was measured with a pain analyzer; the expressions of Beclin-1 and Bcl-2 mRNA and protein in the spinal cord (including nerve root) were detected by fluorescence quantitative PCR and immunohistochemistry, separately. The autophagosome and ultrastructure of the spinal nerve root tissue were observed by transmission electron microscope. RESULTS: After modeling, the gait score was significantly increased (P<0.01) and the pain threshold significantly decreased (P<0.01) in the model group in comparison with the blank group. There was no statistical difference in Beclin-1 and Bcl-2 mRNA and protein expression between the blank and the model groups. After intervention, compared with the model group, the gait scores were significantly reduced (P<0.01), the pain threshold and the expressions of Beclin-1 and Bcl-2 mRNA and protein were significantly increased (P<0.01，P<0.05) in the mild moxibustion and mild moxibustion+3-MA groups. The improvement of the above indicators as more significant in the mild moxibustion group than that in the mild moxibustion+3-MA group (P<0.05). After modeling, the organelles in the spinal nerve root tissue cells of the model group were damaged and there were a small amount of autophagosomes. Compared with the model group, the ultrastructure of the spinal nerve root tissue cells in the mild moxibustion group were relatively complete, and the number of autophagosomes increased. CONCLUSION: Mild moxibustion at GV14 has a good analgesic effect on CSR rats, which may be related to its effects in up-regulation of Beclin-1 and Bcl-2 expressions and activation of autophagy and inhibition of apoptosis.