Objective To observe the specific changes of content of cyclic guanosine monophosphate (cGMP) around meridian skin and specific influences of electroacupunture Neiguan (PC6) on cGMP of rats with acute myocardial ischemia (AMI). Methods With 6 male Wistar rats as normal group, 18 rats were made AMI model and divided into model group, high-frequency group and low-frequency group (n=6). The high-frequency and low-frequency group were given electroacupuncture in bilateral PC6 with 100 and 2 Hz for 20 min respectively. The content of cGMP in skin around PC6, Ximen (PC4), Tianquan (PC2), Waiguan (TE5) and Zusanli (ST36) was detected by using enzyme-linked immunosorbent method after treatment. Results In model group, cGMP content increased significantly around PC6, PC2, TE5 and ST36 compared with normal group (P <0.05,P <0.001). After the treatment with high-frequency or low-frequency electroacupunture in PC6, cGMP content decreased significantly in skin around PC6, PC2, TE5 and ST36 compared with model group (P <0.05, P <0.001). Conclusion High-frequency electroacupunture or high-frequency electroacupunture used in PC6 can protect rats from the injury of AMI. The mechanism may be related to the decrease of cGMP content around the acupoint areas in heart-related meridian.

Objective To illustrate the composition ratio of ERβ isoforms in paired cancerous and adjacent normal tissues from breast cancer patients.Methods Eighty-seven pairs of cancerous and adjacent normal tissues were obtained from breast cancer patients.RT-qPCR was used to determine the relative mRNA expression levels of ERβ isoforms (ER[β1,ERβ2 and ERβ5),and the composition ratios of ERβ isoforms were analyzed.Results The expression levels of all tested ERβ isoforms (ERβ1,ERβ2 and ERβ5) in breast cancer tissues were much lower than those in adjacent normal breast tissues (P ＜ 0.01).Isoform ratio analysis showed that ERβ5 was the dominant isoform in both cancerous and adjacent normal tissues with a positive detection rate of 54.02 ％ and 75.84 ％,respectively.Meanwhile,ERβ1 had the lowest detection rate (9.74 ％ and 6.77 ％ in cancerous and adjacent normal tissues,respectively).The positive rates for both ERβ1 and ERβ2 were much lower in adjacent normal tissues than those in cancer tissues (Z =-2.24,P =0.025 and Z =-4.85,P ＜ 0.01,separately),while more cancerous tissues were ERβ5-positive in comparison to adjacent normal tissues (Z =-5.32,P ＜ 0.01).Conclusions The expression levels of all the ERβ isoforms are differentially down-regulated with significant alterations in their composition ratios during breast carcinogenesis.Further understanding on molecular mechanisms underlying the differential down-regulation of ER[β isoforms will shed new light on breast carcinogenesis.

Objective:To study the effect of microRNA (miR)-330-3p on hepatic ischemia-reperfusion injury (IRI) in mice, meanwhile, and to determine potential molecular mechanism.Methods:Eighty male C57BL/6 mice, aged 7-8 weeks, 23-25 g, specific pathogen free, were randomly divided into 8 groups (10 mice in each group) using random number table: reperfusion 2 h group, 6 h group, 12 h group, 24 h group, sham group, miR-330-3p agomir group (preoperative injection of agonist), miR-330-3p antagomir group (preoperative injection of inhibitor) and miRNA-NC group. Except for the sham group, the hepatic IRI model were established in mice. Polymerase chain reaction (PCR), Western blot and immunohistochemistry were used to detect the expression of miR-330-3p and phosphoglycerate mutase family member 5 (PGAM5), cleave caspase-1 and GSDMD. Luciferase reporter assay was performed to investigate whether miR-330-3p directly targets PGAM5. At the same time, AML12 cells were also treated with miR-330-3p mimics/inhibitor or PGAM5 siRNA, then the expression of PGAM5, NLRP3, cleave caspase-1 and GSDMD were detected by Western blot analysis.Results:Level of miR-330-3p gradually decreased after reperfusion, however, mRNA level of PGAM5 was increased thereafter ( P<0.05) as compared with the sham group. Serum level of AST and ALT were decreased in miR-330-3p agomir group while that of were increased in miR-330-3p antagomir group as a function of time following reperfusion, and the differences were statistically significant (all P<0.05). Cleave caspase-1 expression was decreased in miR-330-3p agomir group but was increased in miR-330-5p antagomir group ( P<0.05). Luciferase reporter assay was performed to determine PGAM5 was a target gene of miR-330-3p. SiRNA-mediated knockdown of PGAM5 decreased level of GAM5 (0.24±0.09), NLRP3(0.12±0.07), cleave caspase-1 (0.15±0.07) and GSDMD (1.08±0.08) as compared with the siRNA-NC group (1.17±0.14), (0.36±0.09), (0.68±0.09), (1.36±0.08), and the differences were statistically significant (all P<0.05). Conclusion:MiR-330-3p can alleviate hepatic IRI in mice, which may be related to inhibition of PGAM5-induced pyroptosis.

Objeetive To analyze the clinical efficacy of liver transplantation (OLT) for patients with hepatopulmonary syndrome (HPS).Methods From 2008 to 2013,420 adult patients underwent liver transplantation in our hospital.There were 91 patients with,and 329 patients without,HPS.The 5-year survival and mortality rates after OLT for the two groups were retrospectively analyzed.Results There were no significant differences between patients without and with HPS in age,primary disease,Child-Pugh score,MELD score,cold ischemia time and warm ischemia time.However,the differences on serum albumin [(29.6 ± 1.2) g/L vs.(26.4 ± 1.6) g/L] and blood oxygen pressure [(61.0 ±9.0) mmHg (1 mmHg =0.133 kPa) vs.(87.0 ± 6.0) mmHg] were significantly different (P ＜ 0.05).The 1-year cure rate was 65.9％ (60/91) in 91 patients with HPS after liver transplantation.The 1,3,5-year cumulative survival rates for patients without HPS were 97.3％,90.9％ and 80.3％,respectively,and the main causes of death were primary graft dysfunction,recurrent cardiovascular events and primary disease recurrence or tumors.The 1,3,5-year cumulative survival rates for patients with HPS were 65.9％,59.3％ and 56.0％,and the main causes of death were multiple-organ failure,pulmonary infection and cerebrovascular events.Kaplan-Meier survival curve analysis showed that the survival of patients with HPS was significantly lower than that of patients without HPS (P ＜ 0.05).Conclusions Liver transplantation is the most effective treatment for patients with HPS,but the short-term mortality rate is relatively high.We still need to learn more about HPS to improve the survival rate of patients with HPS after liver transplantation.

Objective To explore the effect of different acupuncture stimuli on uterine micro-circulation of dysmenorrheal rats with cold stagnation syndrome. Methods Totally 32 three-month old female SD rats in diestrus were randomly divided into saline control group, model group, A stimuli group, and B stimuli group, 8 rats in each group. Model group and treatment groups were given whole body freezing combined with estradiol benzoate injection method to establish models. A stimuli group was given deep puncture with manipulation, while B stimuli group was treated by shallow puncture without manipulation. Diameter of uterine capillary,micro-vessel, TXB2, and 6-keto-PGF1αlevels were observed in each group. Results Compared with the saline group, capillary diameter in model group was significantly reduced at 5, 10, 20, 30 min time point (P<0.01);micro-vascular diameter was significantly reduced at 5, 10, 20, 30, 40 min time point (P<0.01);plasma 6-keto-PGF1α levels decreased (P<0.01);TXB2/6-keto-PGF1αincreased significantly (P<0.01). Compared with model group, capillary diameter in A stimuli group enlarged at 5, 10, 20, 30 min time point (P<0.05), micro-vascular diameter dilated at 5, 10, 20, 30, 40 min time point (P <0.01), plasma 6-keto-PGF1α level increased (P <0.05), TXB2/6-keto-PGF1αdecreased significantly (P<0.05);micro-vascular diameter in B stimuli group dilated at 20, 30 min time point (P<0.05), plasma TXB2/6-keto-PGF1α decreased significantly (P<0.05). Compared with B stimuli group, capillary diameter in A stimuli group dilated at 5, 10, 20, 30 min time point (P<0.05) and micro-vascular diameter dilated at 5, 10, 20, 30, 40 min time point significantly (P<0.01). Conclusion Dysmenorrheal rats with cold stagnation syndrome show obvious disorder of the uterus micro-circulation and circulation related substances. Both A and B acupuncture stimuli improved uterus micro-circulation of dysmenorrheal rats with cold stagnation syndrome, and its mechanism may be related to the recovery the balance between TXB2 and 6-keto-PGF1α.

Objective Infrared thermal imaging can be applied to the diagnosis and auxiliary diagnosis of some diseases . The aim of this study is to explore acupuncture-induced changes in skin temperature in acupoint areas and whether skin temperature in -creases or decreases in the acupoint areas along meridians . Methods Thirty two female SD rats were randomly and equally divided into four groups:saline control,cold congealing and dysmenorrhea model , Sanyinjiao (SP6), and Guanyuan( CV4).Models were es-tablished in the latter three groups by subcutaneous injection of estradiol benzoate at 0.5 mg for 10 successive days and , 1hour after the last administration , intraperitoneal injection of oxytocin at 2 U, followed by exposure of the rats to-25℃in a freezer 4 hours a day for 5 days.Meanwhile , the control rats received normal saline only and were not exposed to low temperature .Infrared thermal imaging was used to measure the skin temperature at the acupoint areas of SP6, Xuehai (SP10), and CV4 before and at 0, 5, 10, 20, 30, 40, 50 and 60 min after needling . R esults At 0 to 5 min after nee-dling, the skin temperature of the left SP6 and right SP10 was signifi-cantly decreased in both the SP6 and CV4 groups ( [ -0. 56 ± 0.22]℃and [-0.48 ±0.11]℃, P<0.01), and so was that of the right SP10 ([ -0.64 ±0.21]℃ and [ -0.45 ±0.13]℃, P<0.05).At 5 to 10min, the skin temperature of the right SP6 and SP10 was markedly increased in the SP6 group ([-0.49 ±0.35]℃and [-0 .18 ±0.20]℃, P<0.01), and so was that of the right SP6 in the SP6 group at 20 to 30 min ([ -0.14 ±0.25]℃) as compared with the model and CV4 groups (P<0.01).At 30 to 40 min, the skin temperature of the right SP10 was remarkably elevat-ed in the SP6 group ([ -0.03 ±0.11]℃) in comparison with the model group (P<0.01).No significant differences were observed in the skin temperature of the left SP10 and CV4 at different time points among the four groups (P>0.05). Conclusion The skin temperature of SP6 and SP10 can be regulated by needling both the acupoints of SP 6 and CV4.The increase in the skin temperature of the right SP6 and SP10 in the SP6 group and no change in the CV 4 group indicated dynamic temperature changes in the acupoint area along the meridian after needling.

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.

Back-shu points were firstly discussed in(the), and various location methods of back-points had been proposed by acupuncturists until conclusive method was made indynasty. In this paper, the different location methods of back-points were reviewed; based on this, the reasons of divergences among each theory on location methods were discussed, and the theoretical background and reference of the original establishment of back-points were further explored. Therefore, it was proposed that the standardized location of back-points should be just considered as the center of possible distribution range, and adjustment should be made during clinical application according to the variability of individual combined with finger pressing.

Objective According investigate the expression of NLRP3 in liver tissues of mice with hepatic ischemia-reperfusion injury (HIRI),to determin the role of NLRP3 in the process of HIRI.Methods Established mice model of partial HIRI.Forty-two male C57BL/6 mice (aged 7 to 8 weeks,weight 20 to 25 g) were respectively divided into 7 groups:no-treatment control group,sham operation group,HIRI groups (2、6、12、24 h) and CY-09 group,6 mice in each group.The injury of the hepatic tissues in the 7 groups was analyzed based on detecting the levels of alanine transaminase (ALT),aspartate transaminase (AST),interleukin-1β (IL-1β),interleukin-18 (IL-18),tumor necrosis factor-α (TNF-α) by ELISA.HE and TUNEL staining were used to observe the pathological changes of liver tissues after HIRI.Western blotting assay were carried out to detect the expressions of NLRP3 and Caspase-1.Measurement data were expressed as mean ± standard deviation (Mean ± SD),and one-way variance analysis was used for comparison between groups.If the variance was not uniform,Dunnett C test was used.Results Serum ALT,AST,IL-1 β,IL-18 and TNF-α of mice detected in HIRI groups were higher than no-treatment control group and sham operation group at all endpoints (P ＜ 0.05).The relative expression of NLRP3 and Caspase-1 in the liver tissues of mice in the HIRI groups were significantly higher than that in the no-treatment control group and sham operation group.Serum ALT,AST,IL-1β,IL-18 and TNF-α of mice detected in CY-09 group were lower than HIRI groups at all endpoints (P ＜ 0.05).Less hepatocellular necrosis were exhibited in CY-09 group,comparing to HIRI groups.The hepatocyte apoptosis rate of mice in the CY-09 group was significantly lower than that in the 12 h HIRI group (P ＜ 0.05).The relative expression of NLRP3 in the liver tissues of mice in the CY-09 group was significantly lower than that in other groups.The relative expression of Caspase-1 in the liver tissues of mice in the CY-09 group was significantly lower than that in other groups except the no-treatment control group and sham operation group.Conclusions HIRI cause an increase in NLRP3 expression.The inhibition of NLRP3 can reduce HIRI.

V set and Ig domain containing 4 (VSIG4), a co-inhibitory molecule expressed by macrophages, is a B7 family-related protein. It can serve as the second signal of T cell activation and regulate the function of T cells. It has been found that VSIG4 is essential in maintaining immune tolerance. Moreover, VSIG4 can also act as a complement receptor, playing a role in recognizing pathogens, regulating the complement alternative pathway and inhibiting inflammatory response. This review summarized the expression of VSIG4 and its role in the immune system.