Objective To explore the effect of electroacupuncture at Hegu (LI 4) on the expressions of P2X2 and P2X3 receptors in experimental pulpalgia rats.Method Forty-two male SD rats were randomized into a normal group (group N), a control group (group C), a pulpalgia model group (group M), an antagonist group (group A), an electroacupuncture group (group E), and an antagonist+electroacupuncture group (group AE), 7 rats in each group. Group N didn’t receive any interventions; group C received injection of normal saline into pulp cavity of the same dose as the injection in group M, and the cavity was then blocked by dental fillings 5-6 min later; in group M, maxillary first and second molar teeth were drilled (drill bit of 1 mm in diameter) to expose pulp and lipopolysaccharide (LPS) solution 5μg/μL was injected into the holes (1~3μL for each hole), and the holes were then covered by dental fillings 5-6 min later; group A received the same modeling method as that in group M, but A-317491 was injected together with LPS (0.5 mg/kg); group E received electroacupuncture at bilateral Hegu (LI 4) with needles retained for 30 min, once a day, totally for 3 times; group AE received the same electroacupuncture intervention after receiving the same treatments as that in group A. The rats’ behaviors and weight were observed for 30 min after intervention each day. The rats were sacrificed on the 4th day, and the mRNA expressions of P2X2 and P2X3 receptors andβ-actin in trigeminal ganglion were detected by using RT-PCR. The mRNA expressions were then compared among the groups.Result The behavioral changes in group M, E, and AE were more significant than that in group C and N (P<0.01); the behavioral changes in group A, E, and AE were less significant than that in group M (P<0.01). The weight in group C was significantly lower than that in group N (P<0.01); the weights in group M, A, E and AE were significantly lower than that in group C and N (P<0.01); the weights in group E and AE were significantly higher than that in group M and A (P<0.01,P<0.05); the weight in group A was slightly higher than that in group M (P<0.05); the weights in group AE was significantly higher than that in group E (P<0.01). The mRNA expressions of P2X2 receptor in group M, A, and AE were significantly higher than that in group N and C (P<0.01); the mRNA expressions of P2X2 receptor in group A, E, and AE were lower than that in group M (P<0.05); the mRNA expression of P2X2 receptor in group A was lower than that in group E (P<0.05); the mRNA expression of P2X2 receptor in group E was higher than that in group AE (P<0.05). The mRNA expressions of P2X3 receptor in group M, A, and E were significantly higher than that in group C (P<0.05) and group N (P<0.01); the mRNA expressions of P2X3 receptor in group A, E, and AE were significantly lower than that in group M (P<0.01); the mRNA expression of P2X3 receptor in group AE was markedly lower than that in group E (P<0.01).Conclusion The expressions of P2X2 and P2X3 receptors in trigeminal ganglion were increased in LPS-induced pulpalgia rats. Electroacupuncture at Hegu (LI 4) and injection of A-317491 both can down-regulate the mRNA expressions of P2X2 and P2X3 receptors, which is plausibly the action mechanism of electroacupuncture at Hegu (LI 4) in analgesia.

OBJECTIVETo determine the regulatory effects of the circadian clock gene Per1 on cell cycle-related genes and its influence on the proliferation, apoptosis, cycle, and tumorigenicity in vivo of human oral squamous cell carcinoma SCC15 cells.

METHODSThree groups of the short hairpin RNA (shRNA) of lentivirus recombinant plasmids were designed against the RNA of Per1 and then transfected to the SCC15 cells. The optimum interference group was screened through Western blot and quantitative real-time PCR (qRT-PCR) and assigned as the experimental group. The transfected lentivirus plasmid without an interference effect on any gene was set as the control group (Control-shRNA). Untreated SCC15 cells were set as the blank group. The mRNA expressions of cell cycle-related genes, namely, Per1, p53, Cyclin D1, Cyclin E, Cyclin A2, Cyclin B1, CDK1, CDK2, CDK4, CDK6, p16, p21, Wee1, cdc25, E2F, and Rbl1 in each group were detected through qRT-PCR. The cell proliferation, apoptosis, and cell cycle distribution in each group were evaluated through flow cytometry. The cells of the experimental group and the blank group were subcutaneously inoculated in nude mice to observe tumorigenesis.

RESULTSThree groups of Per1-shRNA lentivirus plasmids were constructed successfully. Among the groups, the Per1-shRNA- I group exhibited the highest interference effect, as indicated by qRT-PCR and Western blot analysis. As such, this group was set as the experimental group. The mRNA expression levels of CyclinD1, CyclinE, CyclinB1, CDK1, and Wee1 gene in the Per1-shRNA-I group were significantly higher than those in the Control-shRNA group and the SCC15 group (P < 0.05). By contrast, the mRNA expression levels of p53, Cyclin A2, p16, p21, and cdc25 in the Per1-shRNA-I group were significantly lower than those in the Control-shRNA group and the SCC15 group (P < 0.05). The mRNA expression levels of each gene between the Control-sLRNA group and the SCC15 group did not significantly differ (P > 0.05). The mRNA expression levels of CDK2, CDK4, CDK6, E2F, and Rb1 did not significantly differed in the three groups (P > 0.05). The proliferation index of the Perl-shRNA-I group was significantly higher than those of the Control-shRNA group and the SCC15 group (P < 0.05). The apoptosis index of the Per1-shRNA-I group was significantly lower than those of the Control-shRNA group and the SCC15 group (P < 0.05). The number of S-phase cells in the Per1-shRNA-I group was significantly lower than those of S-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). The number of G2/M-phase cells in the Per1-shRNA-I group was significantly higher than those of G2/M-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). Conversely, the proliferation index, apoptotic index, and cell cycle distribution of the cells in the Control-shRNA group did not significantly differ from those of the SCC15 group (P > 0.05). The tumorigenic ability in vivo was significantly enhanced in the Per1-shRNA-I group (P < 0.05).

CONCLUSIONPer1 is an important tumor suppressor gene. Per1 can regulate a large number of downstream cell cycle-related genes. The alteration of its expression can affect cell cycle progression, proliferation, apoptosis imbalance, and tumorigenic ability in vivo. Further studies on Per1 may elucidate cancer development and provide novel effective molecular targets for cancer treatment.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Circadian Clocks ; genetics ; Cyclin D1 ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; Period Circadian Proteins ; genetics ; Plasmids ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Transfection

Objective To investigate the immunological reaction mechanism of medicated serum in Zibai Gelatin for SiHa cells of the cervical cancer infected by high risk human papilloma virus (HPV). Methods Through immunohistochemical comparison and the cell culture, and after medicated serum was administrated to SiHa cells of cervical cancer for 24 h, 48 h, and 72 h, optical densities of IL-6, IL-10, CD83 and TNF-αin the same time but different concentrations and different times but the same concentration were observed. Relationships of dose-effect and time-effect between expressions of IL-6, IL-10, CD83 and TNF-α and medicine action were analyzed by calculating average optical density. Results With the increase of medicine concentration and administration time of Zibai Gelatin, the expressions of IL-6 and TNF-αgradually decreased, while the expressions of CD83 and IL-10 gradually increased (P<0.05). Conclusion Zibai Gelatin with medicated serum can inhibit local inflammatory reaction by improving local immunologic function of cervix, which is beneficial to reduce cervix high-risk HPV.

Objective To introduce a new method performing umbilicus abdominal access during laparoscopy. Methods In a prospec-tive,362 patients received laparoscopic appendectomy in October 2012 to October 2014 were oberserved in this study. The clinical data,the time of the progress of making umbilicus abdominal access,complications and the state of the wound were recorded. Results The performing time of the trocar in umbilicus was (42. 3 ± 27. 1)s. Conclusion The method Cut-open,is simple,fast and save,which is worth to promoting in laparoscopic surgery.

Objective to investigate the efficacy of moxibustion treatment to rheumatoid arthritis(RA) patients and the impact to blood levels of rheumatoid factor(RF) ,C reactive protein(CRP) ,and interleukin-6(IL-6) .Methods RA patients were divided in-to treatment group and comfort group .The two groups were treated by moxibustion and sham-moxibustion for 2 courses ,respec-tievely .Results Overall clinical efficacy rate was higher in treatment group than in comfort group(P<0 .01) .The arthralgia ,ar-throcele ,tenderness and its degrees ,and morning stiffness time were significant improved in both groups(P<0 .05) ,and the indexes were better in treatment group than comfort group(P<0 .05) .The blood levels of RF ,IL-6 and CRP were reduced after treatment in both groups(P<0 .05) ,and which of treatment group reduced more than those of comfort group(P<0 .05) .Conclusion Moxi-bustion treatment can significantly improve clinical outcomes of RA and markedly reduce the blood levels of RF ,IL-6 and CRP of RA patients .

Objective Investigate the effect of EC50 of propofol target-injection in the different TCM (traditional Chinese medicine) physique types of patients who are in unconscious phase. Methods Depend on the standard protocol of TCM physique types sort and determination, we sorted 60 patients into three groups:Ping He (group A), Yang Xu (group B) and Yin Xu (group C), 20 patients per group. We applied the sequential experiment to measure the minimal EC50 and NI values of propofol when the patients were in the unconscious phase. Results The EC50 of propofol of group A, B and C are 3.85 μg/mL, 4.12 μg/mL and 3.43 μg/mL respectively. 95% confidence intervals of group A, B and C are 3.64 ~ 4.07 μg/mL, 3.92 ~ 4.33μg/mL and 3.26 ~ 3.60 μg/mL respectively. Conclusion There is a correlation between the different TCM physique types and the dosage of propofol target-injection.

Aim To study the effect of Macrophage colony-stimulating factor(M-CSF) on MMP-9 in RAW 264.7 cell and explore the relationship between atherosclerosis caused by M-CSF and the activity of MMP-9. Methods Gelatin zymography analysis was used to investigate the effect of M-CSF and PD98059 on the activity of MMP-9 in cultured RAW 264.7 cell.Western blot was used to study the effect of M-CSF and PD98059 on the express of p-ERK1/2 in cultured RAW 264.7 cell. Results The enzyme activity of MMP-9 was significantly increased after 24-hour M-CSF treatment.Meanwhile, M-CSF upregulated the expression of p-ERK1/2. Pre-treatment with PD98059 blocked partly the increased expression of p-ERK1/2 and the activity of MMP-9 induced by M-CSF. Conclusion M-CSF can induce the secretion of MMP9 in RAW 264.7 cell, which may be mediated by the phosphorylation of ERK1/2.

Objective To report a modified radical inguinal lymphadenectomy the aim of which is to reduce the incidence of the complication of skin necrosis.Methods One hundred and twenty-six modified radical inguinal dissections were performed in 63 patients with penile carcinoma from June 2002 to June 2010.A modified radical inguinal dissection characterized by an S-shaped incision and precisely separating layers using an anatomical mark was performed.The boundaries of dissection were the same as classic radical inguinal lymphadenectomy.The incidences of complications of skin flap necrosis were retrospectively analyzed.Results The follow-up time ranged from 12 to 93 months.A total of 37 complications occurred,including 7 minimal skin necroses (5.6％),3 wound infections (2.4％),19 lymphedemas (15.1％),2 seroma formations (1.6％),5 lymphocele (4.0％),and 1 deep venous thrombosis (0.8％).There were no complications in 106 dissections (84.1％ ),and 20 dissections had one or two minor complications ( 15.9％ ).Conclusions The complication of skin necrosis related to groin dissection in patients with penile carcinoma could be reduced by using this modified inguinal dissection technique,which characterized by an S-shaped incision and precisely separating layers with using an anatomical landmark.

Objective To investigate the penetrative ability of amlodipine gels and to evaluate their effect on the survival of rat dorsal ischemic random skin flaps.Methods 0.5 ％,1.0 ％,1.5 ％,2.0 ％,and 2.5 ％ amlodipine gels were made.The accumulative penetrative quantities of amlodipine through rat skin were assayed with a modified Franz's diffusion cell in vitro.Pure gel without amlodipine,0.5 ％ and 1.5 ％ amlodipine gel were respectively applied on rat random ischemic skin flap once a day for 7 days.The viable area was measured on the seventh postoperative day and the quantities of amlodipine within skin flap were also detected at 2 and 6 hours after application of amlodipine gel.Results The accumulative penetrative quantities of amlodipine increased in time-and concentration-dependent manner (P＜0.05).Accumulative quantities of 0.5 ％ and 1.0 ％ amlodipine gel were lower than those of 1.5 ％,2.0 ％,and 2.5％ gel,respectively (P＜0.05).The quantities of amlodipine within flap tissue in 1.5 ％ amlodipine gel was higher than that of 0.5 ％ amlodipine gel (P＜0.05).The survival area of flap in 0.5 ％ amlodipine gel group (391.4±65.4) mm2 was higher than those of the pure percutaneous gel group (192.9±56.8) mm2 and the control group (191.0±50.2) mm2(P＜0.05),but no significant difference was seen between 1.5 ％ amlodipine gel group (265.7+88.3) mm2 and control group (P＞0.05).Conclusions Amlodipine could penetrate into skin tissues.0.5 ％ amlodipine gel could significantly increase survival area of ischemic random skin flap.

Abortion, Spontaneous ; diagnosis ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; Diagnosis, Differential ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hydatidiform Mole ; diagnosis ; genetics ; metabolism ; Immunohistochemistry ; Pregnancy ; Uterine Neoplasms ; diagnosis ; genetics ; metabolism