BACKGROUND: Using the rapid growth period of the chorioallantoic membrane from the 8th day to the 12th day, vascular growth at bone defect end and vascular implantation of a bone graft substitute into the human body can be simulated.OBJECTIVE: To load hydroxyapatite into a chick embryo chorioallantoic membrane model, and to establish an implanted revascularization model.METHODS: Twelve rosette eggs were incubated under the same suitable conditions for 8 days. After windowing, the eggs were randomly divided into two groups: experimental group was implanted with hydroxyapatite material to establish the model of chick embryo chorioallantoic membrane carrying hydroxyapatite; control group was implanted with filter paper material, sealed and placed into a constant temperature and humidity box to continue to hatch. At the 12th day of incubation, length of new vessels and length of new vessels per unit area in the two groups were observed, and the angiogenesis on the hydroxyapatite and filter paper was observed.RESULTS AND CONCLUSION: The mean length of new vessels and length of new vessels per unit area were 24.031 mm and 0.242 mm/mm2 in the experimental group, and 23.561 mm and 0.212 mm/mm2 in the control group, respectively. There were no significant differences between two groups. Additionally, vascular tissues were obviously observed on the hydroxyapatite samples in the experimental group. These experimental results show that the establishment of the model of chick embryo chorioallantoic membrane carrying hydroxyapatite provides a simple and accurate in vitro animal model for studies on ideal bone graft materials and angiogenesis. The hydroxyapatite has no effect on the angiogenesis on the chorioallantoic membrane.

Objective To compare the effects and explore mechanism of acupoints on various meridians or nerve segments in treating different symptoms in one disease.Methods Neonatal Wistar rats were randomly divided into 4 groups: normal control group, model group, ST36 group, and LI14 group (n=10-12 in each group).IBS model was established by administering acetic acid enema combining colorectal distension (CRD) test to rats in all groups except the normal control group.These rats were all isolated from their mothers.2-month old rats in the ST36 group and LI14 group received electro-acupuncture five times, once every two days for 20 minutes.Stool consistency was evaluated according to the Bristol Stool Form Scale (BSFS), and visceral sensitivity was observed based on abdominal withdraw reflection (AWR).Levels of TRPV1 and 5-HT2AR in rat colon tissues were measured with immunohistochemical method.Results Before electro-acupuncture stimulation, BSFS score of rats in all groups increased significantly (P<0.01) compared with the normal control group;after electro-acupuncture, BSFS score of the model group (P<0.01) and LI14 group (P<0.05) were increased;threshold value of intestinal pressure of model group and ST36 group decreased (P<0.01), and the contraction waves increased significantly (P<0.01).Positive TRPV1 expression increased (P<0.01);and TRPV1 expression in LI14 group increased (P<0.05).The level of 5-HT2AR in the model group and LI14 group were raised markedly (P<0.01).Compared with the model group, BSFS score of the EA groups decreased significantly (P<0.01);threshold value of intestinal pressure increased (P<0.01);contraction wave decreased significantly (P<0.01), and the expression of TRPV1 and 5-HT2AR reduced significantly (P<0.01).Compared with the ST36 group, BSFS score of the LI14 group increased significantly (P<0.05);expression of TRPV1 reduced (P<0.01), and the level of 5-HT2AR increased significantly (P<0.01).Conclusion Stimulating LI4 and ST36 with electro-acupuncture could reduce the visceral sensitivity and the gastrointestinal motility disorder in IBS model rat.LI4 has a better effect of treating gastrointestinal motility disorder, and ST36 has a better effect of relieving visceral pain.This study illustrates the existence of therapeutic specificity of acupoints, which means acupoints on different meridians or ganglions can be used to treat the same disease yet with different effects.

Objective To observe the analgesic effect of acupuncture at Guanyuan (CV4) and its effect on vasomotor substances in rats with dysmenorrhea due to coagulated cold syndrome. Method The coagulated-cold dysmenorrhea rat model was developed by Estrodiol benzoate and Oxytocin injectin plus physical freezing. The writhing response (writhing latency, writhing frequency, and writhing score) was observed, and the contents of TXB2 and 6-keto-PGF1a were detected by using enzyme-linked immunosorbent assay (ELISA). Result Compared with the saline water group, the writhing latency was significantly shortened, the writhing frequency was significantly increased, and the writhing score was more significantly increased in the model group (P<0.01);compared with the model group, the writhing latency was significantly prolonged, the writhing frequency was decreased, and the writhing score was significantly lower in the acupuncture group (P<0.05, P<0.01). Compared with saline water group, the content of plasma 6-keto-PGF1a was significantly lower (P<0.05) and the content of plasma TXB2 showed an increasing tendency (P>0.05) in the model group. Compared with the model group, the content of plasma 6-keto-PGF1a showed an increasing tendency (P>0.05) and the content of plasma TXB2 showed a decreasing tendency (P>0.05) in the acupuncture group. Conclusion The vasomotor substances are obviously disordered in the blood of cold-coagulated dysmenorrhea rat models. Acupuncture at Guanyuan can improve the writhing response and release pain, and meanwhile positively regulate the vasomotor substances such as TXB2 and 6-keto-PGF1a. The vasomotor substances are plausibly one of the major substances in the action of acupuncture in preventing and treating dysmenorrhea.

Objective To observe and compare the effects of LI 4 and ST 36 electroacupuncture on co-lonic motility and positive expression of M3 receptor in colon of irritable bowel syndrome ( IBS ) model rats, and to explore the mechanism behind those differences. Methods 40 neonatal Wistar rats were randomly divided into blank control group, model group, LI 4 group, and ST 36 group. IBS rat model was built by using neonatal maternal separation and acetic acid enema, and colorectal distension ( CRD) stimulation in rats except the blank control group. At 9 weeks rats in LI 4 group and ST 36 group were given electroacupuncture( EA ) intervention once every other day for 5 times. Blank group and model group didn' t receive acupuncture intervention. After the treatment, Bristol Stool Scale was applied to i-dentify the feces types of the IBS rats before and after electroacupuncture invention;Abdominal withdraw-al reflex ( AWR) was evaluated to asess the intestinal sensitivities and smooth muscle contraction intensi-ty;and latency, contraction wave and pressure value of initial wave were observed;The expression of M3 receptor was measured using immunohistochemistry. Results Bristol scores of LI 4 group and ST 36 group significantly decreased (P<0. 01). Compared with blank group, the stool score of model group was significantly increased ( P<0 . 01 );the latency was shortened ( P<0 . 01 ); the number of contrac-tion waves was significantly increased (P<0. 01);the immunohistochemical expression of M3 receptor in the colon was significantly increased (P<0. 01). The immunohistochemical expression of M3 receptor in the colon in LI 4 group was increased (P<0. 01). Compared with model group, after electroacupuncture stimulation, the stool score of LI 4 group and ST 36 group was significantly decreased (P<0. 01); the latency was significantly prolonged ( P <0 . 01 ); the number of contraction wave was significantly de-creased in LI 4 group (P<0. 01) and ST 36 group (P<0. 05);immunohistochemical expression of M3 receptor in the colon was significantly decreased (P<0. 01). Compared with the LI 4 group, the stool score in ST 36 group was decreased ( P<0 . 05 );the pressure value of initial wave significantly decreased (P <0. 01); the immunohistochemical expression of M3 receptor in the colon was decreased ( P <0. 05 ) . Conclusion Stimulating LI 4 or ST 36 with electroacupuncturing could both inhibit gastrointesti-nal tract contraction, reduce intestinal sensitivity and relieve IBS symptoms in rats. LI 4 appears to be more effective than ST 36 in relieving abdominal pain, while ST 36 is better than LI 4 to stop diarrhea. Stimulating different acupoints or acupoints on different nerve segments with electro-acupuncture have specific effects on IBS rats.

Objective To investigate the expression,synergistic relationship and clinical significance of alcohol de-hydrogenase(ADH1A)and vascular endothelial growth factor-A(VEGFA)in hepatocellular carcinoma(HCC).Methods The expression and correlation of ADH1A and VEGFA in HCC and adjacent normal tissues were ana-lyzed by GEPIA.TCGA and GSEA were used to analyze the pathway of ADH1A in HCC.The clinical and patho-logical data of 84 patients with HCC were collected,and 54 patients with paracancer normal tissue samples were se-lected as controls to analyze the correlation between ADH1A and VEGFA and clinicopathological parameters of HCC.Immunohistochemistry was used to detect the protein expression of ADH1A and VEGFA in cases and con-trols,and the correlation between the expression of ADH1A and VEGFA and the clinical progression and prognosis of patients with HCC was analyzed based on clinical pathological parameters and Kaplan-Meier.Results Bioinfor-matics analysis found that ADH1A was low-expressed in HCC and VEGFA was highly expressed in HCC,and there was a negative correlation between the two(P<0.001);immunohistochemical detection results showed that the expression of ADH1A in HCC tissue was lower than that in normal tissue adjacent to cancer(P<0.01)while the expression rate of VEGFA in HCC tissue was significantly higher than that of normal tissue adjacent to cancer(P<0.01);The recurrence rate of vascular thrombus and HCC patients in HCC group with high expression of ADH1A was lower(P<0.05).The proportion of tumor diameter>5 cm,high TNM stage,microsatellite and G2-G3 dif-ferentiation in HCC tissues in VEGFA high expression group was higher(P<0.05).Kaplan-Meier survival analy-sis showed that patients with high ADH1A expression and low VEGFA expression had a higher five-year survival rate.Conclusion Low expression of ADH1A and high expression of VEGFA in tumor tissues of patients with HCC indicate tumor progression and can be used as one of the prognostic evaluation indicators for patients with HCC.

Objective:To discuss the effect of sialic acid-binding immunoglobulin-like lectin-15(Siglec15)derived from M2 tumor-associated macrophages(M2-TAMs)on promoting the malignant biological behavior of the esophageal squamous cell carcinoma(ESCC)through bioinformatics analysis,and to validate the findings through cell experiment.Methods:The Tumor Immune Estimation Resource(TIMER)online Database was used to analyze the expression differences and immune infiltration of Siglec15 in pan-cancer and adjacent normal tissues.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Siglec15 mRNA in M2-TAMs and ESCC EC109 and KYSE150 cells.Based on the non-contact co-culture of M2-TAMs and ESCC cells,the following groups were set up,such as EC109/KYSE150 group,EC109/KYSE150+si-NC group(transfected with si-NC sequence),and EC109/KYSE150+si-Siglec15 group(transfected with si-Siglec15#1 and si-Siglec15#2 sequences).CCK-8 method was used to detect the proliferation activities of the cells in various groups;wound healing assay was used to detect the wound healing rates of the cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups.Results:The bioinformatics analysis results showed that compared with adjacent normal tissue,the expression levels of Siglec15 mRNA in pan-cancer tissues such as esophageal cancer,colon cancer,and head and neck squamous cell carcinoma tissues were increased(P＜0.05 or P＜0.01),and the expression level of Siglec15 mRNA in esophageal cancer tissue was significantly positively correlated with the infiltration of the macrophages(P＜0.05).Compared with the EC109 cells and KYSE150 cells,the expression level of Siglec15 mRNA in M2-TAMs was significantly increased(P＜0.01).There was no significant difference in the proliferation rate of the cells among EC109/KYSE150 group,EC109/KYSE150+si-NC group,and EC109/KYSE150+si-Siglec15 group(P＞0.05).Compared with EC109/KYSE150 group,after treated for 24 and 48 h,the wound healing rate of the cells in EC109/KYSE150+si-NC group was increased(P＜0.01),the numbers of migration and invasion cells were increased(P＜0.05),and the apoptotic rate was decreased(P＜0.01).Compared with EC109/KYSE150+si-NC group,the wound healing rates of the cells in EC109/KYSE150+si-Siglec15#1 group and EC109/KYSE150+si-Siglec15#2 group were decreased(P＜0.05),the numbers of migration and invasion cells were decreased(P＜0.05),and the apoptotic rates of the cells had no significant difference(P＞0.05).Conclusion:Siglec15 derived from M2-TAMs may be a key factor in promoting the migration and invasion of the ESCC cells.

The researches on the acupoint effect specificity were summarized to explore whether the integrality of the acupoint effect specificity existed and analyze the connection between the relativity and integrality of the acupoint effect specificity. The literature on the clinical and experimental researches relevant with the acupoint effect specificity was retrieved through CNKI from January 2007 to October 2017. A total of 39 papers met the retrieving criteria. Separately, in terms of the holism of TCM theory, the holism of meridian theory and modern research, the integrality of acupoint effect specificity was analyzed. The relativity and the integrality are indicated in the acupoint effect specificity. The integrality of acupoint effect specificity is closely related to the holism of TCM theory as well as the meridians. Just because of its integrality, the acupoint effect specificity is relative, rather than absolute.

OBJECTIVETo compare the effects of electroacupuncture (EA) at "Dachangshu" (BL 25) or "Tianshu" (ST 25) for visceral sensitivity, gene expression product c-kit of colonic Cajal interstitial cells (ICC) and capsaicin receptor 1 (TRPV1) of irritable bowel syndrome (IBS) rats, so as to investigate the effect and mechanism differences of EA at the back point and the front point of large intestine for IBS rats.

METHODSForty-two Wistar neonatal rats were randomly divided into a blank group (9 rats) and a model group (33 rats). IBS model was established with mother and child separation, acetic acid enema in young rats and colorectal dilatation method. Twenty-seven IBS rats in life were randomly divided into a model control group, a Dachangshu group and a Tianshu group, 9 rats in each group. EA (disperse-dense wave, 2 Hz/100 Hz, 0.1-0.3 mA) for 20 min was used at "Dachangshu" (BL 25) and "Tianshu" (ST 25) respectively in the Dachangshu and Tianshu groups, once every other day, totally 5 times. The rats in the model control group were fixed with soft cloth sleeve for 20 min, without acupuncture. No intervention was used in the blank group. The stool property Bristol grading score was recorded before and after intervention in each group. The visceral sensitivity was evaluated by abdominal withdrawal reflex. The latency until the first systolic wave occurred and the number of systolic wave within 90 s were observed. Immunohistochemical was used to detect the positive expressions of c-kit and TRPV1, the ICC colon specific marker.

RESULTSCompared with the blank group, the Bristol score increased，latency period shortened, systolic wave number increased, c-kit and TRPV1 positive expressions increased in the model control group (all <0.01). Compared with the model control group, the Bristol score decreased, latency period increased, systolic wave number decreased, c-kit and TRPV1 positive expressions decreased after intervention in the Dachangshu and Tianshu groups (<0.05, <0.01). Compared with the Dachangshu group, the TRPV1 positive expression decreased after intervention in the Tianshu group (<0.05).

CONCLUSIONEA at "Dachangshu" (BL 25) or "Tianshu"(ST 25) can improve the diarrhea in IBS model rats, reduce the visceral sensitivity, and its mechanism may be related to regulating the expressions of colon c-kit and TRPV1. EA at "Tianshu" (ST 25) is more apparent for TRPV1 than at "Dachangshu" (BL 25).