A multifunctional non-viral vector for the delivery of MTH1-targeted CRISPR/Cas9 system for non-small cell lung cancer therapy.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system adapted from bacteria is a programmable nuclease-based genome editing tool. The long-lasting effect of gene silencing or correction is beneficial in cancer treatment. Considering the need to broaden the practical application of this technology, highly efficient non-viral vectors are urgently required. We prepared a multifunctional non-viral vector that could actively target tumor cells and deliver CRISPR/Cas9 plasmids into nuclei of cancer cells. Protamine sulfate (PS) which contains nuclear localization sequence was utilized to condense plasmid DNA and facilitate nuclei-targeted delivery. Liposome-coated protein/DNA complex avoided the degradation of nuclease in blood circulation. The obtained PS@Lip/pCas9 was further modified with distearoyl phosphoethanolamine-polyethylene glycol-hyaluronic acid (HA) to endow the vector ability to actively target tumor cell. Results suggested that PS@HA-Lip could deliver CRISPR/Cas9 plasmids into nuclei of tumor cells and induce genome editing effect. With the disruption of MTH1 (mutT homolog1) gene, the growth of non-small cell lung cancer was inhibited. Moreover, cell apoptosis in tumor tissue was promoted, and liver metastasis of non-small cell lung cancer (NSCLC) was reduced. Our study has provided a therapeutic strategy targeting MTH1 gene for NSCLC therapy. STATEMENT OF SIGNIFICANCE: CRISPR/Cas9 as a powerful tool for genome editing has drawn much attention. The long-lasting effect possesses unique advantage in cancer treatment. Non-viral vectors have high loading capacity, high safety and low immunogenicity, playing an important role in CRISPR/Cas9 delivery. In our study, a multifunctional non-viral vector for the efficient delivery of CRISPR/Cas9 plasmid was constructed. With the active targeting ligand and nuclei-targeting component, the cargo was efficiently delivered into cell nuclei and exerted genome editing effect. By using this vector, we successfully inhibited the growth and induced the apoptosis of non-small cell lung cancer by disrupting MTH1 expression with good safety. Our work provided an efficient non-vial vector for CRISPR/Cas9 delivery and explored the possibility for cancer treatment.

Resveratrol ameliorates oxaliplatin-induced neuropathic pain via anti-inflammatory effects in rats.

Oxaliplatin (OXA) is a common chemotherapy drug and exhibits clinical activity in several cancer types. Its anticancer clinical effect is frequently accompanied by neurotoxicity. The symptoms include paresthesia and pain, which adversely affect the quality of life of patients. In the present study, five consecutive intraperitoneal injections of 4 mg/kg OXA were used to mimic chemotherapy in rats. OXA administration induced mechanical allodynia, activated spinal astrocytes and triggered the inflammatory response. To explore potential therapeutic options for OXA-induced neuropathic pain, resveratrol (Res) was intrathecally injected into the spinal cord of OXA-treated rats. Paw withdrawal threshold values of OXA-treated rats were increased, indicating an antinociception effect of Res on OXA-induced pain. Additionally, Res treatment reduced the levels of glial fibrillary acidic protein, TNF-α, IL-1ß and NF-κB, which were upregulated in OXA-treated rats (compared with control). Furthermore, Auto Dock data showed that Res binds to cyclooxygenase-2 (COX-2) through six hydrogen bonds. Western blot analysis and reactive oxygen species (ROS) assays indicated that Res treatment decreased COX-2 expression and suppressed ROS production. In summary, intrathecal injection of Res reduced the spinal COX-2-mediated ROS generation and inflammatory reaction, suppressed astrocytic activation, and alleviated OXA-induced neuropathic pain.

Acupuncture for chronic sciatica: protocol for a multicenter randomised controlled trial.

BACKGROUND: Chronic Sciatica is a disabling condition causing considerable medical, social and financial implications. Currently, there is no recognised long-term effective treatment to alleviate sciatica. Acupuncture has been widely used for treating chronic pains with persistent analgesic effects. We aim to evaluate the efficacy and safety of acupuncture for chronic sciatica with follow-up in 52 weeks. METHODS AND ANALYSIS: This is a multicenter randomised sham-controlled trial. A total of 216 patients with chronic sciatica will be enrolled and randomly assigned to the acupuncture or sham acupuncture group. There will be 10 treatment sessions applied in 4 weeks with frequency decreased over time. Patients will complete follow-ups during 52 weeks. The primary outcomes are changes in leg pain intensity and disability from baseline to week 4. Secondary outcomes include back pain intensity, frequency and bothersomeness, quality of life, and global perceived effect. Adverse events will be recorded in detail. ETHICS AND DISSEMINATION: Ethical approval of this trial was granted from the ethics committee of Beijing University of Chinese Medicine and all study centres (No. 2020BZYLL0803). Written informed consent will be obtained from enrolled patients. Trial results will be disseminated in peer-reviewed publications. TRIAL REGISTRATION NUMBER: ChiCTR2100044585 (Chinese Clinical Trial Registry, http://www.chictr.org.cn, registered on 24 March 2021); preresults.

[Effects of bosutinib on the malignant behavior of thyroid papillary carcinoma B-CPAP cells and its mechanisms].

Objective: To investigate the effects of bosutinib on the malignant behavior of thyroid papillary carcinoma B-CPAP cells and its possible mechanisms. Methods: Thyroid papillary carcinoma B-CPAP cells were cultured in vitro with a concentration gradient of(1、2、3、4 and 5 µmol/L)bosutinib intervened for 24 hours, DMSO was used as the control group. Five parallel compound holes were set in each group. Cell counting kit (CCK-8 method) method was used to detect cell proliferation. Transwell assay and cell wound healing assay were used to detect cell invasion and migration. TUNEL staining assay and flow cytometry were used to detect cell apoptosis. Western blot was used to detect the expressions of autophagic proteins (Beclin-1, LC3, p62) and signal pathway proteins (SIK2, p-mTOR, mTOR, p-ULK1, ULK1). Results: Compared with the control group, the cell proliferation activity, migration ability and invasion ability were decreased (P＜0.01), while the cell apoptosis rate was increased (P＜0.01) in the bosutinib concentration groups of 2, 3, 4 and 5 µmol/L . In the concentration groups of 4 and 5 µmol/L, the expression of Beclin-1 (P＜0.05), LC3- Ⅱ/LC3- Ⅰ (P＜0.05), SIK2 (P＜0.01) and p-ULK1 (P＜0.01) protein was decreased, while the expression of p62 (P＜ 0.05) and p-mTOR (P＜0.01) protein was increased. Conclusion: Bosutinib may inhibit the autophagy of thyroid papillary carcinoma cells through SIK2-mTOR-ULK1 signaling pathway to inhibit their proliferation, invasion and migration and promote apoptosis, thereby weakening their malignant behavior.

Coadministration of iRGD peptide with ROS-sensitive nanoparticles co-delivering siFGL1 and siPD-L1 enhanced tumor immunotherapy.

The continuous activation and expansion of tumor-specific T cells by various means are the main goal of cancer immunotherapy. Tumor cells overexpress fibrinogen-like protein 1 (FGL1) and programmmed death-ligand 1 (PD-L1), which respectively bind to lymphocyte-activation gene 3 (LAG-3) and programmmed death-1(PD-1) on T cells, forming important signaling pathways (FGL1/LAG-3 and PD-1/PD-L1) that negatively regulate immune responses. In order to interfere with the inhibitory function of FGL1 and PD-L1 proteins, we designed a new type of reactive oxygen species (ROS)-sensitive nanoparticles to load FGL1 siRNA (siFGL1) and PD-L1 siRNA (siPD-L1), which was formed from a stimuli-responsive polymer with a poly-l-lysine-thioketal and modified cis-aconitate to facilitate endosomal escape. Moreover, tumor-penetrating peptide iRGD and ROS-responsive nanoparticles were co-administered to further enhance the delivery efficiency of siFGL1 and siPD-L1, thereby significantly reducing the protein levels of FGL1 and PD-L1 in tumor cells. Our findings indicated that the dual delivery of FGL1/PD-L1 siRNA was a new and powerful treatment method, which was characterized by increasing the infiltration of effector CD4+ and CD8+ T cells, effectively alleviating the tumor immunosuppressive microenvironment. These findings also supported the superiority and feasibility of nanoparticle-mediated tumor immunotherapy, and may provide a different perspective for cancer treatment. STATEMENT OF SIGNIFICANCE: In addition to the idea that cancer vaccines can promote T cell immune responses, nanoparticle delivery modulators (such as small interfering RNA (siRNA) targeting immunosuppressive pathways) may provide more information for the research of nanoparticle-mediated cancer immunotherapy. In this study, we designed a new intelligent nano-delivery system for co-delivery of siFGL1 and siPD-L1, and demonstrated the ability to down-regulate the expression levels of FGL1 and PD-L1 proteins in tumor cells in vitro and in vivo. The constructed nanoparticle had a good tumor microenvironment responsiveness, and the delivery efficiency was enhanced by co-injection with tumor penetrating peptide iRGD. This project proposed a new strategy for tumor immunotherapy based on smart nano-delivery systems, and explored more possibilities for tumor therapy.

Clinical characteristics and risk factors for symptomatic venous thromboembolism in hospitalized COVID-19 patients: A multicenter retrospective study.

BACKGROUND: High incidence of asymptomatic venous thromboembolism (VTE) has been observed in severe COVID-19 patients, but the characteristics of symptomatic VTE in general COVID-19 patients have not been described. OBJECTIVES: To comprehensively explore the prevalence and reliable risk prediction for VTE in COVID-19 patients. METHODS/RESULTS: This retrospective study enrolled all COVID-19 patients with a subsequent VTE in 16 centers in China from January 1 to March 31, 2020. A total of 2779 patients were confirmed with COVID-19. In comparison to 23,434 non-COVID-19 medical inpatients, the odds ratios (ORs) for developing symptomatic VTE in severe and non-severe hospitalized COVID-19 patients were 5.94 (95% confidence interval [CI] 3.91-10.09) and 2.79 (95% CI 1.43-5.60), respectively. When 104 VTE cases and 208 non-VTE cases were compared, pulmonary embolism cases had a higher rate for in-hospital death (OR 6.74, 95% CI 2.18-20.81). VTE developed at a median of 21 days (interquartile range 13.25-31) since onset. Independent factors for VTE were advancing age, cancer, longer interval from symptom onset to admission, lower fibrinogen and higher D-dimer on admission, and D-dimer increment (DI) ≥1.5-fold; of these, DI ≥1.5-fold had the most significant association (OR 14.18, 95% CI 6.25-32.18, p = 2.23 × 10-10 ). A novel model consisting of three simple coagulation variables (fibrinogen and D-dimer levels on admission, and DI ≥1.5-fold) showed good prediction for symptomatic VTE (area under the curve 0.865, 95% CI 0.822-0.907, sensitivity 0.930, specificity 0.710). CONCLUSIONS: There is an excess risk of VTE in hospitalized COVID-19 patients. This novel model can aid early identification of patients who are at high risk for VTE.

Efficacy of acupuncture for sciatica: study protocol for a randomized controlled pilot trial.

BACKGROUND: Acupuncture is widely used for pain diseases while evidence of its efficacy for sciatica is insufficient. We aim to explore the feasibility and efficacy of acupuncture with different acupoint selecting strategies for sciatica induced by lumbar disc herniation. METHODS: This is a multicenter, three-arm, patient-assessor-blinded randomized controlled pilot trial. Ninety patients will be assigned randomly into 3 groups including disease-affected meridians (DAM) group, non-affected meridians (NAM) group, and sham acupuncture (SA) group in a 1:1:1 ratio. The trial involves a 4-week treatment along with follow-up for 22 weeks. The primary outcome is the change of leg pain intensity measured by the visual analogue scale (VAS) from baseline to week 4 after randomization. Secondary outcomes include functional status, back pain intensity, and quality of life. Adverse events will also be recorded. DISCUSSION: The results will inspire the optimal acupuncture strategy for sciatica and help establish a better design as well as power calculation for a full-scale study. TRIAL REGISTRATION: ChiCTR2000030680 (Chinese Clinical Trial Registry, http://www.chictr.org.cn , registered on 9 March 2020).

Stimuli-responsive nanoparticles for the codelivery of chemotherapeutic agents doxorubicin and siPD-L1 to enhance the antitumor effect.

Cancer cells have been reported to exhibit high resistance against immune system recognition through various cell intrinsic and extrinsic mechanisms. Considerable challenges have been encountered in monotherapy with chemotherapeutics to attain the desired antitumor efficacy. In this study, a nanodelivery system was designed to incorporate doxorubicin (DOX) and programmed death-ligand 1 (PD-L1) small interfering RNA (siRNA), that is, siPD-L1. DOX and siPD-L1 were formed from a stimuli-responsive polymer with a poly-L-lysine-lipoic acid reduction-sensitive core and a tumor extracellular pH-stimulated shedding polyethylene glycol layer. The codelivery system was stable under physiological pH conditions and demonstrated enhanced cellular uptake at the tumor site. Moreover, the combined treatment of DOX and siPD-L1 exhibited improved antitumor effect in vitro and in vivo compared with either modality alone. The combination of chemotherapy and immunotherapy presented in this work through the codelivery of a chemotherapeutic agent and a gene-silencing agent (siRNA) may provide a new strategy for cancer treatment.

Doxorubicin and siRNA-PD-L1 co-delivery with T7 modified ROS-sensitive nanoparticles for tumor chemoimmunotherapy.

Tumor cells avoid immunosurveillance during the tumorigenesis, metastasis and recurrence periods thanks to the overexpressed immunosuppressive molecules on their surface. For instance, the programmed cell death 1 ligand (PD-L1) binds with the T-cells' programmed cell death receptor 1 (PD-1) impairing the anti-tumor activity of the host T cells. In this study, a new reactive oxygen species (ROS) responsive nanoparticle (NP), modified with the HAIYPRH (T7) peptide, was developed for the co-delivery of siRNA-PD-L1 and doxorubicin (Dox). These NPs can block the inhibitory signal responding to T cells and enhance cytotoxicity of Dox against tumor cells. The T7 modification binds to the overexpressed transferrin receptor on tumor cells facilitating its cellular uptake. Dox rapid release is then triggered by the high tumor cells cytoplasmic concentration of ROS, leading to cell apoptosis. Our results demonstrated these NPs exhibited a T7-mediated cellular uptake and an intracellular ROS-triggered payloads release in vitro. They also suggested an improved in vivo 4T1 tumor targeting efficiency and chemoimmunotherapy. Most notably, the co-delivery system exhibited a significantly enhanced antitumor effect over Dox-only loaded NPs following prompting the proliferation of T cells by siRNA-PD-L1. In conclusion, these ROS-responsive NPs provided a promising strategy to combine siRNA-PD-L1 immunotherapy and Dox chemotherapy.

Up-regulation of microRNA-136 induces apoptosis and radiosensitivity of esophageal squamous cell carcinoma cells by inhibiting the expression of MUC1.

OBJECTIVE: This present study is performed to figure out the role of microRNA-136 (miR-136) in radiosensitivity of esophageal squamous cell carcinoma (ESCC) cells through the regulation of MUC1. METHODS: Seventy-four ESCC patients were divided into radiotherapy sensitive group and radiotherapy resistance group. Colony formation assay and flow cytometry were used to test the radiosensitivity of radiotherapy resistant strain and parent strain. The expression of miR-136 between radiotherapy resistant strain and parent strain was detected by RT-qPCR, and the expression of miR-136 in Eca109 and TE-1 cells as well as Eca109-R and TE-1-R cells was detected after different doses of X-ray irradiation. Eca109 and TE-1 cells as well as Eca109-R and TE-1-R cells with overexpression of miR-136 or co-overexpression of miR-136 and MUC1 were constructed. Cell proliferation, colony formation and apoptosis was detected by CCK-8 assay, colony formation assay, and flow cytometry, respectively. RESULTS: The expression of miR-136 in ESCC tissues was lower and MUC1 mRNA and protein expression was higher than that in adjacent normal tissues. The expression of miR-136 was negatively correlated with the expression of MUC1 mRNA in ESCC. Low expression of miR-136 and high expression of MUC1 were associated with tumor size, lymph node metastasis and distant metastasis. The expression of miR-136 increased while the expression of MUC1 decreased in the radiotherapy sensitive group of ESCC patients relative to the radiotherapy resistant group. The colony formation ability of radiation resistant cell line was stronger than that of parent cell line, and the apoptosis rate showed an opposite trend. Up-regulation of miR-136 reduced the survival rate, suppressed colony formation ability and induced apoptosis of ESCC cells under irradiation, which was reversed by upregulated MUC1. CONCLUSION: This study demonstrates that up-regulation of miR-136 induces apoptosis and radiosensitivity of ESCC cells by inhibiting the expression of MUC1.

[Efficacy of Modified Acupuncture Method at Renying (ST 9) for Patients with Cervical Spondylosis of Vertebral Artery Type and Its Impact on Velocity of Cervical Blood Flow].

OBJECTIVE: To observe the clinical efficacy of modified acupuncture at Renying point (ST 9) for patients with cervical spondylosis of vertebral artery type and its influence on velocity of cervical blood flow. METHODS: Fifty-nine cases of vertebral artery type cervical spondylosis were randomly divided into control group (n=30) and treatment group (n=29). Both groups were acupunctured at ST 9, with routine acupuncture technique used in the control group and modified technique in the treatment group, respectively. All cases received two courses of treatment, each course covered consecutive 6 once-per-day treatments. Before and after treatment, transcranial Doppler (TCD) was used to measure the systolic peak blood flow velocity (Vs) of left vertebral artery (LVA), right vertebral artery (RVA) and basilar artery (BA), and the scores of "cervical vertigo symptoms and functional assessment scale" (CVSFAS) were also assessed, separately. RESULTS: CVSFAS scoring, Vs of LVA, RVA and BA after treatment showed significant improvement compared with those before treatment (P<0.01, P<0.05). The efficacy of the treatment group in the above mentioned indexes was superior to that of the control group (P<0.05). The total effective rate of the treatment group was 93.1% (27/29), superior to 70.0% (21/30) of the control group (P<0.05). CONCLUSIONS: The modified acupuncture method at ST 9 is clinically effective in the treatment of cervical spondylosis of vertebral artery type via increasing the Vs of vertebral-basilar artery, improving the local blood circulation and relieving pain.

[Observation on clinical therapeutic effect of acupuncture treatment on functional dyspepsia based on syndrome differentiation].

OBJECTIVE: To observe the clinical efficacy difference in treatment of functional dyspepsia (FD) between syndrome differentiation based acupuncture and ordinary acupuncture. METHODS: Seventy FD patients were assigned to a syndrome differentiation based acupuncture group (Group A) and an ordinary acupuncture group (Group B) by Excel Software randomization. Zhongwan (RN12 ), Tianshu (ST25), and Zusanli (ST36) were needled as main points for patients in Group A. Meanwhile, different combined acupoints were needled according to syndrome differentiation. Only the same main points were needled for patients in Group B. All patients were needled once per day, 30 min each time, 6 days as one treatment cycle, 2 treatment cycles in total. Fasting serum levels of gastrin (GAS) and motilin (MTL) were determined before treatment and after 2 treatment cycles. 36-item Short-form Heath Survey (SF-36) and Nepean Dyspepsia Index [NDI, including Nepean Dyspepsia Symptom Index (NDSI) and Nepean Dyspepsia Life Quality Index (NDLQI)] were assessed before treatment, after 2 treatment cycles, and one month after treatment. RESULTS: Compared with before treatment in the same group, serum levels of GAS and MLT increased in the two groups after 2 treatment cycles (P <0. 05), but changes were more obvious in Group A (P <0. 05). Compared with before treatment in the same group, SF-36 and NDLQI score increased, and NDSI score decreased in the two groups after 2 treatment cycles and 1 month after treatment (all P <0. 05). Compared with Group B, SF-36 and NDLQI score increased in Group A after 2 treatment cycles and 1 month after treatment (P <0. 05, P <0. 01). But NDSI score at 1 month after treatment was lower in Group A than in Group B (P <0.01). CONCLUSION: Syndrome differentiation based acupuncture could evidently improve dyspeptic symptoms of FD patients, and significantly improve their quality of life with remarkable curative effect.

[Effects of syndrome-differentiation acupuncture on life quality in patients with functional dyspepsia].

OBJECTIVE: To observe effects of syndrome-differentiation acupuncture on life quality in patients with functional dyspepsia (FD) in order to evaluate its clinical efficacy. METHODS: One hundred and five cases of FD were randomly divided into a syndrome-differentiation acupuncture group, a regular acupuncture group and a non-acupoint group, 35 cases in each one. Zhongwan (CV 12), Tianshu (ST 25), Zusanli (ST 36) were selected as main acupoints in the syndrome-differentiation acupuncture group. After syndrome differentiation, Danzhong (CV 17) and Zhangmen (LR 13) were added for those with stagnation of liver qi; Pishu (BL 20) and Weishu (BL 21) were added for those with deficiency of spleen-stomach qi; Qimen (LR 14) and Taichong (LR 3) were added for liver-qi invading stomach and Yinlingquan (SP 9) and Neiting (ST 44) were added for dampness-heat blocking stomach. The selection of acupoints in the regular acupuncture group was the same as main acupoints in the syndrome-differentiation acupuncture group. The points 10 mm lateral to the main acupoints were selected in the non-acupoint group. The treatment was given once a day, six days as a treatment course and totally two courses were required. The symptom total score, health-related quality of life survey (SF-36) and Nepean dyspepsia index (NDI) were evaluated before and after the treatment as well as one month after the treatment (follow-up visit), respectively. The efficacy was also assessed. RESULTS: After the treatment, the total effective rate was 87.5% (28/32) in the syndrome-differentiation acupuncture group, which was superior to 74.2% (23/31) in the regular acupuncture group and 20.7% (6/29) in the non-acupoint group (P < 0.05, P < 0.01). Compared before the treatment, the SF-36, NDI and symptom total score after the treatment and in the follow-up visit were all obviously improved in the syndrome-differentiation acupuncture group and regular acupuncture group (all P < 0.05), which was the most obvious in the syndrome-differentiation acupuncture group [after the treatment, SF-36: 84.54 +/- 5.93 vs 81.44 +/- 6.22, 63.46 +/- 6.59; NDSI: 18.94 +/- 9.30 vs 21.23 +/- 8.39, 43.93 +/- 11.26; NDLQI: 71.42 +/- 7.23 vs 63.11 +/- 7.06, 54.87 +/- 6.00; symptom total score: 22.06 +/- 15.80 vs 32.52 +/- 16.88, 47.97 +/- 10.92]; the improvement in the regular acupuncture group was more obvious than that in the non-acupoint group (P < 0.01, P < 0.05). Compared before the treatment, only NDSI score was improved in the non-acupoint group after the treatment (P < 0.05). CONCLUSION: The syndrome-differentiation acupuncture could obviously improve patient's life quality in the treatment of FD, which is an effective therapy for FD.

[Clinical observation on therapeutic effect of electroacupuncture at Quchi (LI 11) for treatment of essential hypertension].

OBJECTIVE: To observe the therapeutic effect of electroacupuncture (EA) at Quchi (LI 11) on blood pressure and blood plasma catecholamines in the patient of essential hypertension and to investigate the mechanism. METHODS: Sixty cases of essential hypertension were randomly divided into an EA group (n=30) and a control group (n=30). In the EA group, bilateral Quchi (LI 11) were selected; and in the control group, western medicine Nicardipine was taken. The variation of blood pressure and blood plasma catecholamines were examined before and after the treatment. RESULTS: (1) After treatment, there were significant reduction in the levels of systolic blood pressure and diastole blood pressure in both groups (P < 0.01); (2) After treatment, significant reduction in levels of adrenaline and noradrenaline were also found in both groups (P < 0.01), however, no significant differences in the level of dopamine were observed in both groups (P > 0.05); (3) The effective rate of 66.7% in the EA group was similar to that of 70.0% in the control group (P > 0.05). CONCLUSION: Both EA at Quchi (LI 11) and western medicine are able to beneficially regulate blood pressure of patients with essential hypertension through adjusting blood plasma catecholamines.

[Therapeutic effects of scalp-acupuncture in patients with vascular dementia induced by cerebral infarction: a randomized controlled trial].

OBJECTIVE: To investigate the effects of scalp-acupuncture on intelligence in patients with vascular dementia (VaD). METHODS: A total of 108 VaD patients were randomly divided into two groups: scalp-acupuncture group and Western medicine group. The scores of Mini-Mental State Examination (MMSE), Blesse Dementia Scale (BDS), Hasegawa Dementia Scale (HDS), Abilities of Daily Life (ADL) before and after treatment in the two groups were detected. RESULTS: A total of 92 patients completed the treatment. The scores of MMSE, BDS and HDS in the two groups were significantly increased, and the ADL scores significantly declined (P<0.05). There was no difference in the scores of MMSE, BDS, HDS and ADL between the two groups before and after the treatment. There was no difference in the total effectiveness between the two groups (P>0.05). CONCLUSION: Scalp-acupuncture therapy could improve the clinical intelligence level of VaD patients.

[Effects of electroacupuncture at "Fenglong" (ST 40) on NO, ET and CGRP levels in the rat with hyperlipidemia].

OBJECTIVE: To probe the regulative effect of electroacupuncture (EA) at "Fenglong" (ST 40) on blood lipids in hyperlipidemia (HLP) rats and the mechanism. METHODS: Eighty Wistar rats were randomly divided into a normal group (fed with basal forage), a model group (fed with high fat forage), an EA group (fed with high fat forage + EA treatment), a western medicine group (fed with high fat forage + Pravastatin sodium). Contents of serum total cholesterol (TC), triacylglycerol (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), endothelin (ET), nitric oxide (NO) and calcitonin gene-related peptide (CGRP) were determined before and after treatment. RESULTS: Compared with the normal group, the body weight, levels of TC, TG, LDL-C and ET were significantly elevated (P < 0.05, P < 0.01) and the levels of HDL-C, NO and CGRP were significantly decreased (P < 0.05) in the model group; compared with the model group, the body weight, levels of TC, TG and LDL-C were significantly decreased (P < 0.01) and the levels of NO and CGRP were significantly increased in the western medicine group and the EA group (P < 0.01, P < 0.05); compared with the EA group, HDL-C level significantly increased in the western medicine group (P < 0.01), and ET level decreased in the EA group and the western medicine group with no significant difference between the two groups (P > 0.05). CONCLUSION: Both EA and Pravastatin sodium have better benign regulative effects on TC, TG, LDL-C, NO and CGRP and can decrease ET level to a certain extent in the rat of hyperlipidemia.

[Effect of electro-acupuncture at Fenglong (GV 16) on nitric oxide and endothelin in rats with hyperlipidemia].

OBJECTIVE: To investigate the effect of electro-acupuncture (EA) at Fenglong (GV 16) on body weight, blood lipids, nitric oxide (NO) and endothelin (ET) in rats with hyperlipidemia (HLP). METHODS: Eighty Wistar rats were randomly divided into normal control group (fed normal diet), untreated group (fed a high-fat diet), EA-treated group (fed a high-fat diet plus EA therapy) and pravastatin-treated group (fed a high-fat diet plus pravastatin tablet). There were 20 rats in each group. The body weight and the blood content of total cholesterol (TC), triacylglycerol (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), NO the and ET of the rats in different groups were measured before experiment and after 30-day treatment. A modified method of cardiac puncture for blood sampling was used for blood collection. RESULTS: Compared with the normal control group, the body weight and the levels of TC, TG, LDL-C and ET in the untreated group were significantly elevated (P<0.01, P<0.05), while the levels of HDL-C and NO were obviously decreased (P<0.05). The body weight and the levels of TC, TG, LDL-C demonstrated significant reduction in pravastatin-treated group and EA-treated group as compared with the untreated group (P<0.01), and the NO content in pravastatin-treated group and EA-treated group was higher than that in the untreated group (P<0.01). Compared with the untreated group, HDL-C level was elevated significantly in pravastatin-treated group, while HDL-C level in EA-treated group was not changed significantly, and there was significant difference in the HDL-C level between pravastatin-treated group and EA-treated group (P<0.01). The level of ET was decreased obviously in pravastatin-treated group (P<0.05), while the level of ET in EA-treated group was not changed significantly (P>0.05). CONCLUSIONS: Both EA therapy and pravastatin have efficient regulation of body weight and the content of TC, TG, LDL-C and NO in HLP rats. To some extent, they are able to regulate the imbalance between ET and NO content under the condition of HLP. Western medicine such as pravastatin can regulate the HDL-C level in HLP rats, while the effect of EA therapy on regulation of the HDL-C level is limited.