Objective To explore the mechanism of the treatment of primary dysmenorrhea(PD)by mild moxibustion on"Shenque"and"Guanyuan"acupoints based on the regulation of cAMP-PKA signaling pathway on TRPV1.Methods Totally 32 female non-pregnant Wistar rats were randomly divided into blank group,model group,mild moxibustion group and capsazepine group,with 8 rats in each group.Except for the blank group,the other groups all used estradiol benzoate intraperitoneal injection combined with ice water bath to establish a PD cold-dampness stagnation syndrome rat model.Intervention began on the first day of modeling,the mild moxibustion group selects"Shenque"and"Guanyuan"for mild moxibustion,20 min per time,the capsazepine group was injected capsazepine 2 mg/kg,once a day for 10 consecutive days.ELISA was used to detect uterine PGF2α and cAMP content,immunofluorescence staining was used to detect TRPV1 expression in uterine tissue,Western blot was used to detect PKA,p-PKA and TRPV1 protein expression.Results Compared with the blank group,the latency period of body twisting in the model group rats decreased,and the body twisting score increased(P<0.01);the contents of PGF2α and cAMP in uterine tissue increased(P<0.01),and the expressions of TRPV1 and p-PKA proteins increased(P<0.01).Compared with the model group,the mild moxibustion group and capsazepine group showed an increase in the latency period of body twisting and a decrease in the body twisting score(P<0.01);the content of PGF2α and cAMP in uterine tissue decreased(P<0.01),and the expressions of TRPV1 and p-PKA proteins decreased(P<0.05,P<0.01).Compared with the mild moxibustion group,the capsazepine group showed an increase in the latency period of body twisting and a decrease in the body twisting score(P<0.01);the contents of PGF2α and cAMP in uterine tissue decreased(P<0.05,P<0.01),and the expressions of TRPV1 protein decreased(P<0.05).Conclusion Mild moxibustion at"Shenque"and"Guanyuan"acupoints has obvious analgesic effect on PD rats,and its mechanism may be related to the regulation of uterine cAMP-PKA signaling pathway mediated TRPV1 protein expression.

Objective:To observe the effects of sparrow-pecking moxibustion at Shenque(CV8)and Guanyuan(CV4)on the writhing reaction and score,the temperature and blood flow perfusion of moxibustion point area and uterus,the serum levels of arginine vasopressin(AVP),prostaglandin(PG)F2α,and thromboxane(TX)B2 in rats with primary dysmenorrhea(PD)due to cold-dampness stagnation,and to explore the possible mechanism of sparrow-pecking moxibustion in treating PD.Methods:Thirty-two healthy non-pregnant female Wistar rats were randomly divided into a normal group,a model group,an ibuprofen group,and a sparrow-pecking moxibustion group,with 8 rats in each group.Except for the normal group,the other three groups were subjected to modeling with cold water bath combined with estradiol benzoate and oxytocin injection.Rats in the sparrow-pecking moxibustion group were treated with sparrow-pecking moxibustion at Shenque(CV8)and Guanyuan(CV4)on the 8th day of modeling,30 min/time,once a day for 3 d;those in the ibuprofen group were treated with 0.8 mL ibuprofen solution(a specification of 125 mg in 10 mL)on the 8th day of modeling,once a day for 3 d;those in the normal group and the model group were given 0.8 mL normal saline,once a day for 3 d.On the 11th day,rats in each group were intraperitoneally injected with oxytocin(2 U/rat),and the writhing incubation period and writhing score in 20 min were observed;the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus in vivo were detected;the serum levels of AVP,PGF2α,and TXB2 were determined.Results:The writhing incubation period was significantly longer(P<0.01)and the writhing score was significantly lower(P<0.01)in the sparrow-pecking moxibustion group and the ibuprofen group than in the model group;compared with the ibuprofen group,the writhing incubation period was prolonged(P<0.01)and the writhing score was decreased(P<0.01)in the sparrow-pecking moxibustion group;compared with the normal group,the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus were significantly decreased,while the serum PGF2α,AVP,and TXB2 levels were significantly increased(P<0.01)in the model group;compared with the model group,the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus were significantly increased,and the serum levels of PGF2α,AVP,and TXB2 were significantly decreased in the ibuprofen group and the sparrow-pecking moxibustion group(P<0.05 or P<0.01);compared with the ibuprofen group,the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus were significantly increased(P<0.05),the serum AVP and TXB2 levels were significantly decreased(P<0.05),while the serum PGF2α level had no statistical difference in the sparrow-pecking moxibustion group(P>0.05).Conclusion:Sparrow-pecking moxibustion had a remarkable analgesic effect on the rats with PD due to cold-dampness stagnation,and the mechanism may be related to the increased temperature and blood perfusion of the moxibustion point area and uterus,as well as the decreased serum PGF2α,AVP,and TXB2 levels.

Objective: To observe the effects of ginger-partitioned moxibustion at Shenque (CV8) and Guanyuan (CV4) on the expression levels of endocrine-related molecules and their receptors in rats with primary dysmenorrhea (PD) due to cold-dampness stagnation, thus to explore their analgesic mechanisms. Methods: Thirty-two female Wistar rats were divided into a normal group, a model group, a ginger-partitioned moxibustion group, and a Western medicine group according to the random number table method, with 8 rats in each group. Except for rats in the normal group, all other rats were treated with oxytocin combined with ice-water bath to establish the rat models of PD due to cold-dampness stagnation. After successful modeling, rats in the normal group and the model group did not receive treatment; rats in the ginger-partitioned moxibustion group received treatments with ginger-partitioned moxibustion at Shenque (CV8) and Guanyuan (CV4); rats in the Western medicine group received ibuprofen by intragastric administration. The writhing response of rats was compared among groups, and the serum levels of prostaglandin F2α (PGF2α), estrogen (estradiol, E2), progesterone (P), and the mRNA expression of PGF2α and E2 receptors in the uterine tissues were detected. Results: No writhing behavior was observed in the normal group; compared with the normal group, the serum PGF2α and E2 levels in the model group were increased (P<0.01), while the P level was decreased (P<0.01), and the mRNA expression levels of the uterine PGF2α and E2 receptors were increased (P<0.01, P<0.05). Compared with the model group, the writhing behavior latency was prolonged, and the writhing response score was decreased in the ginger-partitioned moxibustion group and the Western medicine group (P<0.01); the serum PGF2α and E2 levels in the ginger-partitioned moxibustion group and the Western medicine group were decreased, while the P level was increased (P<0.05 or P<0.01); the mRNA expression levels of the uterine PGF2α and E2 receptors in the ginger-partitioned moxibustion group and the Western medicine group were decreased (P<0.05). Compared with the Western medicine group, the ginger-partitioned moxibustion group showed a prolonged writhing behavior latency, reduced writhing response score (P<0.05), and decreased serum E2 level (P<0.05), while no statistical differences in the serum PGF2α and P levels, or the mRNA expression levels of uterine PGF2α and E2 receptors (P>0.05).Conclusion: The analgesic effect of ginger-partitioned moxibustion on PD due to cold-dampness stagnation may be related to regulating the mRNA expression levels of PGF2α and E2 receptors in the uterine tissues.

AIM: To investigate the effect and mechanism of ursolic acid on high glucose and high-fat injury of human aortic endothelial cells. METHODS: MTT method was used to select the appropriate injury concentration of high glucose and sodium palmitate and UA pre incubation concentration. The levels of NO and ROS, the release rate of lactate dehydrogenase and the activities of antioxidant enzymes including total superoxide dismutase, catalase and glutathione peroxidase were detected by kit. The expression of endothelial nitric oxide synthase, intercellular adhesion molecule, vascular cell adhesion molecule caspase-1 and GSDMD were detected by Western blot. The protective effect of UA on HAEC was observed. Hoechst33342 combined with PI fluorescence staining was used to detect the whole state of cell membrane to explore the occurrence of pyroptosis. RESULTS: Pre-incubation with UA (1 and 5 μmol/L) could reduce the damage of HAEC caused by high glucose and high fat (30 mmol/L Glu + 0.1 mmol/L SPA), enhance HAEC activity, increase NO release and eNOS protein expression, alleviate oxidative stress injury, reduce the protein expression of adhesion molecules and reduce the occurrence of pyroptosis. CONCLUSION: UA may reduce the damage of endothelial cells by inhibiting the oxidative stress response and the occurrence of pyroptosis induced by high glucose and high fat.

The integrated curriculums of basic medicine in Shanxi Medical University are as follows: nine basic medical courses were integrated into 10 medical modules, with additional training including PBL and TBL case discussion, clinical clerkship, flipped classroom and other non-integrated subjects etc. In order to assess the interest of students for integrated curriculum, their intrinsic motivation, as well as their comprehension and application of medical knowledge, we gave anonymous questionnaire to 149 students, 16 teachers in basic medicine, and 10 teachers in clinical medicine. Results showed that more than 90% students were willing to take the integrated curriculum and participate in PBL and TBL case discussion, and students who were unwilling to take the curriculum were less than 10%, they thought that the knowledge of new curriculum system was incoherent. The proportion of students from higher grade who were unwilling to participate in the flipped classroom was increased from 6.7% to 95.0%. Most of the teachers both in basic and clinic medicine believed that new curriculum system was helpful for students to comprehend basic medical knowledge and strengthen their discriminating ability, but did not function in improving students' practical ability. In the further reform on teaching, details like the coherence of knowledge and the content selected when self-studying should be improved.

In order to explore the effect of flipped classroom in theoretical teaching of biochemistry,lipids metabolism was selected to carry out four stages flipped classroom.The teaching effect of flipped classroom was evaluated by test and questionnaire survey of students.The results indicated that most undergraduate students could accept novel teaching method.However,the consciousness and the ability of students' independent learning need to be further improved.It also takes time for the students to adapt to the transition from traditional teaching to flipped c1assroom.In summary,the specificity of Biochemistry may limit the carrying out of flipped classroom to some extent.Therefore,which kind of teaching method is adopted depends on the biochemistry theoretical content.Our work may provide some references for further application of flipped classroom in biochemistry teaching.

Objective To investigate the association of ulcerative colitis (UC) with fork head/ winged helix transcription factor-3 (Foxp3) polymorphisms in Han population in Zhejiang province,China.Methods A total of 381 UC patients and 490 healthy controls were enrolled in this study.The four single nucleotide polymorphisms (SNPs) of Foxp3 (rs3761547,rs2232365,rs2294021,rs3761548) were examined by SNaPshot.The analyses of linkage disequilibrium (LD) and haplotype were also performed in all study subjects.Results When male and female UC patients were compared with their corresponding controls respectively,the alleles and genotypes of the four SNPs were not statistically different (all P ＞0.05).According to severity and location of the disease,the UC patients were divided into different subgroups.The alleles (C,G,A) of (rs2232365,rs2294021,rs3761548) were more frequent in male patients with severe UC than in the male controls (69.6％ vs 34.3％,P =0.001;69.6％ vs 34.3％,P =0.001;39.1％ vs 14.4％,P =0.002,respectively).As compared with the female controls,the alleles (C,G,A) and genotypes (TC + CC,AG + GG,CA + AA) of (rs2232365,rs2294021,rs3761548) were significantly increased in the female patients with severe UC (51.9％ vs 38.0％,63.5％ vs 39.2％,53.8％ vs21.4％,80.8％ vs57.7％,84.6％ vs58.4％,76.9％ vs34.7％,all P＜0.05).The four SNPs above were shown to be in a strong LD both in male and in female subjects.When male and female UC patients were compared with their corresponding controls respectively,nevertheless,each haplotype frequency was not statistically different (all P ＞ 0.05).Conclusions Foxp3 (rs2232365,rs2294021,rs3761548) variations might engender the increased risk of severe UC in Chinese Han patients.

OBJECTIVETo compare the effects of acupuncture, electroacupuncture (EA) and moxibustion on functional constipation in rats.

METHODSSixty male Sprague-Dawley rats were divided into a control group (=8), a model group (=11), a medication group (=8), an acupuncture group (=11), an EA group (=11) and a moxibustion group (=11) by random number table. The rats in the model group, medication group, acupuncture group, EA group and moxibustion group were treated with intragastric administration of loperamide hydrochloride for 6 days continuously to establish the functional constipation models, while equal volume of drinking water was administrated to rats in the control group at the same time. The rats in the acupuncture group, EA group and moxibustion group were respectively treated with acupuncture, EA and moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) one hour after intragastric administration; rats in the medication group were treated with intragastric administration of cisapride suspension. All the treatment was given once a day for 6 days. At the last day of intervention, the 24-hour food intake, stool quantity and its water content were measured in each group; the pushing rate of intestine was measured; the structure of colon tissue and acidic mucus in its mucous layer were observed by hematoxylin-eosin dyeing and alcian blue dyeing; the expression of stem cell factor (SCF) and c-kit mRNA was detected by real-time PCR.

RESULTSCompared with the control group, the 24-hour food intake and stool quantity were reduced in the model group (both<0.01), and the water content of stool and pushing rate of intestine were reduced (both<0.01); compared with the model group, the stool quantity and its water content were increased in the medication group, acupuncture group, EA group (<0.05,<0.01), which were not significantly different from those in the moxibustion group (both>0.05). The pushing rate of intestine in each intervention group was increased (all<0.01). The 24-hour food intake and stool quantity in the medication group were not significantly different from those in the acupuncture group, EA group and moxibustion group (all>0.05), and the water content of stool was only reduced in the moxibustion group (<0.01). The pushing rate of intestine in the acupuncture group and moxibustion group was lower than that in the medication group (both<0.01), while that in the EA group was not significantly different from that in the medication group (>0.05). The water content of stool in the moxibustion group was lower than that in the acupuncture group and EA group (both<0.01). The pushing rate of intestine in the acupuncture group and moxibustion group was lower than that in the EA group (both<0.01). The HE staining result indicated the structure of colon tissue was normal, complete and similar in each group; the alcian blue staining indicated the acidic mucosubstance in the model group was lower than that in the control group; compared with the model group, the acidic mucosubstance in the medication group, acupuncture group, EA group and moxibustion group was all increased. Compared with the control group, the expression of SCF and c-kit mRNA was reduced in the model group (both<0.05); compared with the model group, the expression of SCF and c-kit mRNA was increased in the medication group, acupuncture group, EA group and moxibustion group (all<0.05); compared with the moxibustion group, the expression of c-kit mRNA was reduced in the acupuncture group and EA group (both<0.05).

CONCLUSIONSAcupuncture, EA and moxibustion all can play a positive regulative role on functional constipation in rats, in which EA has the best efficacy, followed by acupuncture.

Aim To investigate the effects of midazolam on porcine isolated coronary artery rings pre-contracted by potassium chloride(KCl)and the possible mechanism.Methods The vessel tension recorder system was used.Isotonic tension of porcine isolated coronary artery rings precontracted by KCl(30 mmol·L-1)was recorded.The vasorelaxing action of midazolam and effects of various drugs were observed in the rings.Results Midazolam(3×10-6～1×10-4 mol·L-1)respectively concentration-dependently reduced the contraction induced by KCl,and there was significant difference between the rings with intact and denude endothelium(P<0.05).On KCl-induced precontraction,midazolam′s relaxation was depressed by L-NAME and the blend of L-NAME and L-Arg(P<0.05),but was not affected by Indo,L-Arg and 1400W.The contraction was not prevented by pretreatment with the inhibitor of Na+/Ca2+ exchanger(KB-R7943).The inhibitor of KATP(Gli)restrained the diastolic function of midazolam(P<0.05),while the inhibitor of BKCa(TEA),Kir(BaCl2),KV(4-AP)had no obvious effect.Conclusions Midazolam produces remarkable vasodilatation on KCl pre-contracted porcine isolated coronary artery rings.Its relaxtion effect is via concentration-dependent and endothelium-dependent mechanisms and relevant to the production of NO.Na+/Ca2+ exchanger is not involved midazolam′s vasodilatation on KCl pre-contracted porcine coronary artery rings.The relaxant mechanism of midazolam may be concerned with KATP.The Kir,BKCa and KV may be not involved.

Aim To explore the effects of chrysin on endothelial dysfunction induced by acute high glucose.Methods ① The effects of chrysin on normal isolated aortic at contraction induced by PE and on endothelial dysfunction induced by high glucose were tested in the following medium: normal group,chrysin group;normal-glucose group: glucose 11mmol·L-1 in Krebs' solution;high-glucose group: glucose 44 mmol·L-1 in Krebs' solution;mannitol group: mannitol 33 mmol·L-1 in Krebs' solution and chrysin group: 44 mmol·L-1 Glu+chrysin 1.0 μmol·L-1 in Krebs' solution.② The effects of chrysin on HUVEC cell viability after incubated in high glucose were observed in the following groups: normal-glucose group: glucose 5.5 mmol·L-1 in culture solution;high-glucose group: glucose 33.3 mmol·L-1 in culture solution;mannitol group: mannitol 27.8 mmol·L-1 in culture solution and chrysin group: chrysin(25,50 μmol·L-1)in culture solution.And the NO release was also testd in these groups.Results ① Chrysin could induce vaso-dilation in a dose-dependent manner at normal glucose.The Emax was(58.94±9.61)％,and the EC50 value was 51.9 μmol·L-1.After incubating the aortic rings with high glucose(44 mmol·L-1)for 4 h,there were significant differences in ACh-induced vascular relaxation between the normal glucose group and the high glucose group.The Emax was(32.12±3.92)％and the EC50 value was 78.0 μmol·L-1 of high glucose group(P<0.01).The endothelium-independent relaxation induced by SNP was not significantly different between the two groups.And chrysin(1.0 μmol·L-1)could reverse the decline of ACh-induced vasorelaxation response induced by high glucose(44 mmol·L-1).The Emax was(70.7±3.87)％and the EC50 value was 0.852 μmol·L-1.② The cell viability of HUVEC was depressed after incubated in high glucose,and chrysin could reverse the decline in a concentration-dependent way.And chrysin in defferent concentrations could increase the cell NO release.Conclusion Chrysin could prevent the acute high glucose-induced vascular endothelial dysfunction and could increase the NO release.