Objective:To observe the analgesia effect and of low-level combined- and single-laser irradiation on Zusanli (ST 36) in rats and the relationship between mast cell degranulation and laser acupuncture analgesia.Methods: A total of 66 Sprague Dawley (SD) rats were randomly assigned into normal control,model control,sham irradiation,10.6 μm laser (220 mW,40 Hz) 650nm laser(36mW,continuous),combined laser (10.6 μm+ 650 nm) groups.Arthritis mode 1 was established by injection of Complete Freund's Adjuvant (0.05 mL) into the left ankle joint.The paw withdrawal latency (PWL) was detected with a radiant heat algesimeter.Zusanli (ST 36) was irradiated with sham,single or combined laser for 30 min.After sacrifice under anesthesia (1% embutal),tissues of Zusanli (ST 36) area were sampled,sliced (5 μm) and stained with Toluidine Blue for skin for observing the mast cell degranulation.Results:Compared with model control and sham group,the pain threshold increased significantly in 650nm and combined laser groups (P＜0.01),while remained no significant difference in 10.6 μm group.Compared with model and sham group,the degranulation ratios of mast cells in 650nm and combined laser group were significantly higher (P＜0.001),while remained no significant difference in 10.6 μm group.The linear correlation coefficient between degranulation ratios of mast cells and PWL after laser acupuncture is 0.737 (P＜0.001).Conclusion:Both 650 nm and combined laser stimulation of Zusanli (ST 36) can effectively raise pain threshold,and enhance degranulation ratio of mast cells at the stimulated acupoint.The result also suggested a linear correlation between degranulation ratio of mast cell and analgesia effect.

Objective: To evaluate the effects of 650 nm-10.6 μm combined laser in patients with knee Osteoarthritis (OA) and to determine whether the combined laser provides greater pain relief and improved function compared with red light. Methods: Forty-eight patients with knee OA were randomly allocated to two groups (24 per group), receiving 20 rain irradiation with 650 nm -10.6 μm combined laser or red light emitting diode respectively on point Dubi (ST 35) 3 times a week for the first course (2 weeks) and twice a week for the second one (4 weeks). The main outcome measures were WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) scores. In addition, patients' global assessment, adverse effects and validation of patient blinding were analyzed. Results: All the patients completed the first course, but 12 were lost during the second one. Due to the high dropout rate by the second course, only the data acquired from the first course could be analyzed. No differences of general data of patients and WOMAC scores were found in between-group comparison before treatment (P>0.05). The WOMAC scores of patients in both combined laser group and red light group reduced significantly compared to baseline by the end of the first course (P<0.01). There were no significant differences on the reduction rate of WOMAC scores between two groups (P>0.05). Neither the patients' global assessment nor the dropout rate showed statistical differences between two groups (P>0.05). There was no difference between two groups in patients correctly guessing the treatment assignment (P>0.05). There was no significant difference in the reduction rate of WOMAC scores and the patients' global assessment between patients who guessed their assignment (P>0.05). Conclusion: Both combined laser and red light irradiation are beneficial to patients with knee OA. But as the statistical indifferences between two groups, the authors can't conclude from this study whether the combined laser is more effective.

Objective To explore the effect of small dose of dexmedetomidine on nausea and vomiting in the patients with cervical cancer minimally invasive surgery .Methods One hundred and twenty-two cases of cervical cancer minimally invasive sur-gery in the hospital from June 2014 to June 2016 were selected as the research subjects and divided into the control group and obser-vation group according to the random number table method ,61 cases in each group .The observation was intravenously injected by 0 .1μg/k dexmedetomidine for 1 min before anesthesia induction ,while the control group was given the same volume of normal sa-line .The perioperative indexes ,nausea and vomiting grade and adverse reactions occurrwence were comparfed between the two groups .The anesthesia time ,operative time ,eye opening time ,autonomous respiratory recovery time and tracheal tube extubation time had no statistical differences between the two groups were compared .Results The intraoperative HR minimum value in the observation group was lower than that in the control group ,the bradycardia occurrence rate was higher than that in the control group ,the difference was statistically significant (P<0 .05) .The constituent ratio of PONV grade Ⅱat T2 time point in the obser-vation group was significantly lower than that in the control group ,and the occurrence rate of nausea and vomiting at T 4(postopera-tive 24 h) in the observation group was significantly lower than that in the control group ,the difference was statistically significant (P<0 .05) .The occurrence rate of agitation in the control group was higher than that in the observation group ,and the difference was statistically significant (P<0 .05) .Conclusion Small dose of dexmedetomidine can significantly alleviate postoperative nausea and vomiting in the patients with cervical cancer minimally invasive surgery ,moreover can decrease the agitation occurrence rate without affecting the effect of anesthesia ,but is prone to develop bradycardia .

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kip1-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endothelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combination was used as negative control (pGenesil-HK). The recombination of four plamids was confirmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kip1 mRNA and p27Kip1 protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group). pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The proliferation rates of the pGenesil-P3 group, the pGenesil-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesil-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kip1 could down-regulate the expression of p27Kip1 effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.
Cell Proliferation ; Cornea/cytology ; Cyclin-Dependent Kinase Inhibitor p27/*metabolism ; Endothelial Cells/*cytology ; Endothelial Cells/metabolism ; Gene Expression Regulation ; Models, Biological ; Plasmids/metabolism ; RNA Interference ; RNA, Messenger/metabolism ; RNA, Small Interfering/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tetrazolium Salts/pharmacology ; Thiazoles/pharmacology ; Transfection

Objective:To explore the effects and mechanism of Salvianolic acid B(SalB)on Parkinson's disease(PD).Methods:Forty-eight C57BL/6 male mice were randomly separated into control group(control)withoutdrugs,model group(MPTP)with intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrah-ydropyridine(MPTP),SalB control group with intraperitoneal injection of SalB,and SalB treatment group(MPTP+SalB).Construction of PD mouse model by intraperitoneal injection of MPTP,and treatment with intraperitoneal injection of SalB.Pole climbing test was applied to assess behavior differences.The time of first black stool excretion and water content of feces were measured to evaluate intestinal dysfunctions.The number of tyrosine hydroxylase(TH)positive cells in substantia nigra and the level of Toll like receptor 4(TLR4)in colon were analyzed by immunohistochemistry.The pathological changes of colonic mucosa were observed by HE staining.The levels of calprotectin(CP)and tumor necrosis factor-α(TNF-α)in colon were determined by ELISA.Western blot was used to determine the level of TH in midbrain,the protein level of TH,tight junction protein(ZO-1),and protein level of TLR4/MyD88/NF-κB signaling pathways which express in colon.Results:Com-pared with the Control group,the climbing time,T-turn time and the first black stool excretion time in MPTP group increased while the fecal water content and the number of TH positive cells in substantia nigra were decreased.Accompanied by TLR4 positive cells in colon,pathological injury score of colonic mucosa,levels of CP and TNF-α in colon increased,expression of TH in midbrain and expression of ZO-1 in colon decreased.Expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon increased.Com-pared with MPTP group,SalB treatment shortened the climbing time,T-turn time and the first black stool excretion time in SalB treat-ment group,increased the fecal water content and the number of TH positive cells in substantia nigra,lowered TLR4 positive cells in colon,enhanced expression of TH in midbrain and colon,reduced the pathological injury score of colonic mucosa,significantly decreased levels of CP and TNF-α in colon,enhanced expression of ZO-1 in colon,inhibited expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon.Conclusion:SalB can protect the nerves and intestines and alleviate the intestinal inflamma-tion of PD mice,which may be related to the inhibition of TLR4/MyD88/NF-κB signal pathway.

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kipl-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endo- thelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combi- nation was used as negative control (pGenesil-HK). The recombination of four plamids was con- firmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kipl was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kipl mRNA and p27Kipl protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group), pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The pro- liferation rates of the pGenesil-P3 group, the pGenesii-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kipl on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesiI-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kipl could down-regulate the expression of p27Kipl effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.

To investigate the mutation site of pathogenic gene in patients with hypertrophic cardiomyopathy (HCM) and to analyze the relationship between the genotype and clinical phenotype. Methods: Targeted exon capture sequencing was conducted in a HCM proband for 30 coding exons related HCM gene by all exon amplification and high-throughput sequencing. Furthermore, Sanger sequencing was performed in other family member and in 200 healthy volunteers for verification. The familial investigation included in clinical presentation, physical examination, electrocardiogram and echocardiography. Results: There were 3/6 blood relatives carrying cardiac myosin-binding protein gene MyBPC3 G772A heterozygous mutation, the mutation site was at 258 amino acid of MyBPC3 as glutamic acid (Glu) was substitute to lysine (Lys), such mutation was not found in rest of family member and not in healthy volunteers. The onset of proband and her daughter was rather late, they had palpitation and chest tightness; echocardiography showed interventricular septum basal segment thickening (16-18) mm. Proband was complicating paroxysmal ventricular tachycardia, malignant arrhythmia and heart failure, the maximum pressure gradient of left ventricular outflow was 56 mmHg, which with the high risk for sudden death. Conclusion: Comprehensive gene test has been helpful for clinical stratification, early diagnosis and treatment. MYBPC3 site mutation c.G772A might be the pathogenic mutation in that specific HCM family.