Objective:To observe the clinical effect of acupuncture therapy to support yang and consolidate the constitution for cervical spondylosis of vertebral artery type (CSA). Methods:A total of 72 patients with CSA were randomly divided upon the random digital table into an acupuncture- moxibustion group and an acupuncture group, 36 cases in each group. The acupuncture-moxibustion group was treated by electroacupuncture (EA) at bilateral Fengchi (GB 20) and C4-6 Jiaji (EX-B 2), in combination of moxibustion at Baihui (GV 20), Shenshu (BL 23) and Mingmen (GV 4) to support yang and consolidate constitution. The acupuncture group was given by routine EA at bilateral Fengchi (GB 20), cervical Jiaji (EX-B 2), Tianzhu (BL 10), Jianjing (GB 21), Houxi (SI 3), Hegu (LI 4) and Waiguan (TE 5). The treatment was given once every day and continuous six sessions made one course. After 2-course treatment, the improved situation of clinical symptoms and clinical effect were observed in the two groups. Results:The total effective rate was 91.7% in the acupuncture-moxibustion group and 61.1% in the acupuncture group. The difference of the total effective rate between the two groups was statistically significant (P<0.05). After treatment, the scores of the clinical symptoms in the two groups were obviously decreased than those of the same group before treatment (P<0.01). The differences in the scores between the two groups were statistically significant (P<0.05). Conclusion:Acupuncture-moxibustion treatment to support yang and consolidate constitution is better than the routine acupuncture therapy in the therapeutic effect for CSA.

Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.

Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.

Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.

Objectives To investigate the prevalence of heterogeneous vancomycin intermediate Staphylococcus aureus(hVISA) and the sensitivity of hVISA to novel antibiotics,and to explore the risk factors and infection attributable mortality associated with hVISA infection.Methods A total of 456 methicillin resistant Staphylococcus aureus (MRSA) isolates were isolated in Zhongshan Hospital from January,2008 to November,2010.All MRSA isolates were investigated for hVISA by two agar screening methods BHIA5T (brain-heart infusion containing teicoplanin 5 mg/L)or BHIA6V (brain-heart infusion containing vancomycin 6 mg/L),as well as macroEtest method(MET).Possible hVISA isolates were tested by modified population analysis profile-area under the curve (PAP-AUC).The minimal inhibitory concentrations(MICs) of vancomycin,teicoplanin and linezolid were determined by microbroth dilution as recommended by Clinical Laboratory Standards Institute(CLSI).The contribution difference between hVISA and vancomycin susceptible Staphylococcus aureus (VSSA) in different MIC range was compared.A retrospective case-control study of the patients with hVISA infection or VSSA infection was carried out and statistical analysis was performed using t test,Mann-Whitney test,x2 test and Fisher exact test.Results A total of 105 isolates of hVISA were screened by BHIA5T and BHIA6V (23.0％) with other 23 isolates by MET(5.0％) and 21 by PAP-AUC(4.6％).All isolates were 100％ sensitive to vancomycin,teicoplanin and linezolid.The vancomycin MIC [(1.76 ±-0.16) mg/L] in hVISA group was significantly higher than that in VSSA group[(1.09 ± 0.07)mg/L,P ＜ 0.01],which was a potential risk factor for hVISA infection.The retrospective study showed chronic obstructive pulmonary disease (COPD) was also a risk factor for hVISA infection of the lower respiratory tract.No significant difference in infection attributable mortality was showed between the hVISA group and the VSSA group.Conclusions The overall prevalence of hVISA in Zhongshan Hospital is estimated as 4.6％,while the prevalence of hVISA isolated from blood is as high as 12.5％.All isolates are 100％ sensitive to vancomycin and linezolid.COPD is a risk factor for hVISA infection of the lower respiratory tract.

Objective To investigate the effects of berberine on tumor-associated macrophages (TAM)and the expression of cyclooxygenase-2 (COX-2)of intestinal polyps in Apc(Min/+) mice.Methods A total of 20 Apc(Min/+) mice,four weeks old,were equally divided into the control group and the berberine group,10 in each group.The mice of the control group drank plain water,while the mice of berberine group drank water with 0.1 % berberine.After 12 weeks,all the mice were sacrificed.The intestine and colon were isolated,and the numbers of polyps were counted.The expression of F4/80,inducible nitric oxide synthase-2 (iNOS),macrophage mannose receptor (MR)and COX-2 was detected by immunohisto-chemistry method.The relative expression of COX-2 at protein level was measured by Western blotting. The t test was performed for comparison between two independent groups.Results The total number of intestinal polyps,the number of small intestinal polyps and the number of colon polyps of the berberine group (11 .50±2.05 ,10.50±1 .77 and 1 .00±0.46,respectively)were all less than those of the control group (30.63±1 .69,28.00±2.00 and 2.63±0.74,respectively),and the differences were statistically significant (t=16.727,16.952 and 3.162,P =0.001 ,0.001 and 0.010,respectively).The percentage of F4/80 positive cells in the stroma of polyps of the berberine group ((17.40 ±4.23 )%)was less than that of the control group ((31 .24±6.34)%),and the difference was statistically significant (t =5 .327, P =0.043).The percentage of iNOS positive cells in the stroma of polyps of the berberine group ((7.43± 1 .78 )%) was higher than that of the control group ((2.72±0.68)%), and the difference was statistically significant (t=7.335 ,P =0.004).The percentage of MR positive cells in stroma of polyps of the berberine group ((19.52±1 .54)%)was less than that of the control group ((12.63±0.68)%),and the difference was statistically significant (t=5 .634,P =0.016).The percentage of COX-2 positive cells in stroma of polyps of berberine group ((3.38 ± 0.51 )%)was less than that of the control group ((7.60±0.57 )%),and the difference was statistically significant (t = 7.234,P = 0.001 ).The relative expression of COX-2 at protein level of polyps of the berberine group was lower than that of the control group. Conclusion Berberine may take the role in inhibiting the growth of intestinal polyps in Apc(Min/+) mice through interfering the differentiation of TAM in polyps and suppression the expression of COX-2.

Objective To evaluate the association of gastric hyperplastic polyps with colorectal neo-plasia.Methods Data of consecutive patients who underwent esophagogastroduodenoscopy (EGD)and colonoscopy between January 2011 and December 2013 were reviewed retrospectively.They were compared with patients without gastric polyps who also underwent EGD in the same period.The relationship between gastric polyps and colorectal neoplasia was analyzed.Results The incidence of colorectal neoplasia in gas-tric hyperplastic polyps group was higher than that of the control group [24.0% (46 /192)VS 10.4%(40 /384),P =0.000].An increased incidence of colorectal adenomas in gastric hyperplastic polyps group was found,but there was no difference in the incidence of colorectal cancer in gastric hyperplastic polyps group and control group[2.1%(4 /192)VS 2.3%(9 /384),P =1.000].Stratification analysis suggested that the incidence of colorectal neoplasia in gastric hyperplastic polyps group who aged over 50 was signifi-cantly higher than that in the control group[28.5%(43 /151)VS 10.6%(29 /274),P =0.017],regard-less of different genders and the number of gastric hyperplastic polyps.Conclusion The incidence of color-ectal neoplasia in gastric hyperplastic polyps has significantly increased,especially in those aged over 50 years,regardless of different genders and the number of gastric hyperplastic polyps.Such patients should undergo colonoscopy to detect colorectal neoplasia.

Objective To observe the effect of acupoint application of Chinese medicine on preventing constipation after stroke. Methods By means of randomized, double-blind, controlled study design, 96 patients of yang-syndrome stroke were randomly divided into observation group and control group, 48 cases in each group. Both groups were given conventional treatment for stroke and routine nursing for preventing constipation, and additionally, the control group was given acupoint application of placebo while the observation group was given acupoint application of self-made umbilical paste of Radix et Rhizoma Rhei(Dahuang) and Natrii Sulfas (Mangxiao). The incidence of constipation, frequency of laxative usage and adverse reactions of the two groups were observed. Results (1) The incidence of constipation of the observation group was lower than the control group on treatment day 7 and 14 , the difference being statistical significant(P < 0.05 or P < 0.01).(2)On treatment day 7 and 14, the Bristol scores of stool properties of observation group were increased compared with those before treatment(P < 0.05), but the changes of scores in the control group were insignificant(P > 0.05). The observation group had better effect on improving stool properties(P < 0.05).(3) The frequency of use of laxatives was reduced in the observation group compared with the control group, the difference being significant(P<0.05).(4) The incidence of adverse reaction was 6.25%in the control group and was 8.333%in the observation group, the difference being insignificant (P>0.05). The adverse reaction of both groups was mild, and had no effect on the accomplishment of the treatment. Conclusion Acupoint application of Chinese medicine is effective and safe on preventing constipation after stroke, showing good prospects for clinical application.

Objective:To investigate the clinical efficacy of dopamine hydrazine combined with carlobidopa controlled-release tablets in the treatment of Parkinson's disease and its effect on inflammatory reaction, serum homocysteine and YKL-40 levels in patients with Parkinson's disease.Methods:A case-control study was conducted to retrospectively analyze the clinical data of 230 patients with Parkinson's disease who received treatment at Xianyang Hospital of Yan 'an University from January 2020 to December 2022. According to the different treatment methods, these patients were divided into a dopamine hydrazine (treatment with dopamine hydrazine, n = 113) and a combination group (treatment with dopamine hydrazine and carlobidopa controlled-release tablets, n = 117). All patients were treated for 6 weeks. Inflammatory reactions, serum homocysteine levels, and serum YKL-40 levels post-treatment were compared with pre-treatment levels. Clinical efficacy and the incidence of adverse reactions were evaluated between the two groups. Results:After 6 weeks of treatment, the levels of tumor necrosis factor α, interleukin-1β, interleukin-6, and high-sensitivity C-reactive protein in the combination group were (9.76 ± 1.57) ng/L, (27.02 ± 3.41) ng/L, (10.29 ± 1.89) ng/L, and (6.62 ± 0.99) mg/L, respectively, which were significantly lower than those in the dopamine hydrazine group [(12.34 ± 1.97) ng/L, (30.84 ± 4.73) ng/L, (13.83 ± 2.18) ng/L, (8.77 ± 1.55) mg/L, t = -10.98, -7.04, -13.16, -12.58, all P < 0.05]. The serum levels of homocysteine and YKL-40 in the combination group were (19.08 ± 5.70) μmol/L and (3.33 ± 0.97) g/L, respectively, which were significantly lower than those in the dopamine hydrazine group [(22.54 ± 5.62) μmol/L, (4.03 ± 1.23) g/L, t = -4.63, -4.81, both P < 0.05]. The total response rate in the combination group was 94.02% (110/117), which was significantly higher than 78.76% (89/113) in the dopamine hydrazine group (χ2 = 11.47, P < 0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Conclusion:The use of dopamine hydrazine combined with carbidopa controlled-release tablets for the treatment of Parkinson's disease reduces inflammatory reactions, decreases serum homocysteine and YKL-40 levels, enhances clinical efficacy, and does not increase the incidence of adverse reactions.

Objective:To explore the effects of the γ-aminobutyric acid(GABA) neurons and melanin-concentrating hormone (MCH) neurons of the nucleus accumbens (NAc)-lateral hypothalamic area (LHA) neural pathway on the rewarding feeding(palatable food sweat condensed milk) in the obesity rats.Methods:Total 142 male Wistar rats of SPF grade were divided into normal diet (ND) group ( n=68) and high-fat diet induced obesity (DIO) group ( n=74) according to the principle of body mass matching. The rats in the two groups were given normal diet and high-fat diet for 8 weeks. Eight weeks later, 6 DIO rats were randomly selected to observe the nerve projection from GABA neurons in NAc to MCH neurons in LHA by fluorogold retrograde tracing combined fluorescence immunohistochemistry. And the expressions of c-Fos and MCH in LHA after ingestion of sweet condensed milk(rewarding feeding) were observed by fluorescence immunohistochemistry (6 rats in each group). GABA receptor agonist Musimol or GABA receptor antagonist Bicuculine was microinjected into the nucleus of LHA to observe the effect of GABA on rewarding food intake in ND and DIO rats ( n=8 in each group), and the changes of rewarding food intake after blocking MCH signal ( n=8 in each group). SPSS 17.0 was used for statistical analysis, two-way ANOVA and post hoc Bonferroni test were used for comparison among multiple groups, and t-test was used for comparison between two groups. Results:After 8 weeks of high-fat diet modeling, the intake of delicious food in DIO rats was significantly higher than that in ND rats((12.52±2.29) mL, (7.45±1.23) mL, t=4.778, P<0.01) after satiety.The results of fluorogold retrograde tracing combined with fluorescence immunohistochemistry showed that GABA neurons in NAc projected nerve fibers to neurons in LHA, and GABA A receptors in some neurons in LHA coexisted with MCH.The results of NAc-LHA pathway on delicious food intake showed that the interaction between rat group and drug intervention was significant( F=9.869, P<0.01). Simple effect analysis showed that the intake of delicious food after microinjection of Musimol into LHA nucleus of ND rats was significantly lower than that of microinjection normal saline ((4.25±1.38) mL, (7.29±1.49) mL, P<0.01), while the intake of delicious food after injection of Bicuculine was significantly higher than that of microinjection normal saline((10.72±2.11) mL, (7.29±1.49) mL, P<0.05). The intake of delicious food after microinjection of Musimol into LHA nucleus in DIO group was significantly lower than that of microinjection normal saline((3.51±1.77)mL, (13.68±2.95) mL, P<0.01), but there was no significant difference between microinjection Bicuculine and microinjection normal saline ((14.83±3.44) mL, (13.68±2.95) mL, P>0.05). The results of blocking MCH signal on delicious food intake showed that the interaction effect between SNAP-94847 and Bicuculine intervention was not significant ( F=1.468, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=15.880, P<0.01)and the main effect of Bicuculine intervention was significant ( F=6.930, P<0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of ND rats was significantly less than that of injection normal saline((4.78±1.72) mL, (7.63±2.77) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((6.24±2.18) mL, (4.78±1.72) mL, P>0.05). In the DIO rats, the interaction effect between SNAP-94847 and Bicuculine intervention was not significant( F=0.006, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=18.46, P<0.01) and the main effect of Bicuculine intervention was not significant ( F=2.059, P>0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of DIO rats was significantly lower than that of injection normal saline((6.89±2.11) mL, (12.19±4.36) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((8.72±2.26) mL, (6.89±2.11) mL, P>0.05). Conclusion:GABAergic signal in NAc can regulate the expression of MCH in neurons of LHA. In the DIO rats, the sensitivity of MCH neurons in LHA to satiety signal decreases and the hedonic feeding increases.