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Objective To compare the efficacy and safety of ultrasound guided pectoral nerves Ⅱ (Pecs Ⅱ) block with thoracic paravertebral nerve (TPVN) block for postoperative analgesia after modified radical mastectomy.Methods Eighty female patients scheduled for radical mastectomy,aged 40-65 years,ASA physical status Ⅰ or Ⅱ,were randomly divided into 2 groups using a random number table method (n =40 each):the patients in group T received TPVN block,whereas the pa tients in group P received Pecs Ⅱ block.Both the groups received 0.5 ％ ropivacaine 25 ml.The blocks were performed under all aseptic precautions in the preoperating room 30 min before surgery.The total number of dermatomes that had less pain to pin prick compared with opposite side were not ed.All patients were observed for 30 min after performing the block.The patients were received patient-controlled intravenous analgesia (PCIA).The duration of analgesia and total analgesic consumption in 24 h after surgery were recorded.Adverse effects were recorded between the two groups.Results The duration of analgesia in group P was significantly prolonged than group T [(326.5± 47.8) min vs (201.4±34.5) min,P＜0.01].The 24 h sufentanil consumption were also decreased in group P [(6.9±1.2) μg vs (10.7±1.9) μg,P＜0.01].T2 dermatomal spread were significantly increased in group P [35 (87.6％) cases vs 9 (22.5％) cases,P＜0.05].No complication was recor ded.Conclusion Ultrasound-guided Pecs Ⅱ block and TPVN provided safe and effective anesthesia in patients undergoing modified radical mastectomy,but the effect of Pecs Ⅱ block were more satisfied and per sistent.
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Objective To construct a CaME141G fusion protein-expressing plasmid,and to express,purify,and identify the activity of the recombinant protein. Methods The 141st site of the wild type CaM,E (GAG),was mutated to G (GGG),using site-specific mutagenesis technology. Escherichia coli BL-21 was transformed with the mutant plasmid. The GST-CaME141G fusion protein was mass-cultured and induced for expression. Subsequently,the GST-CaME141G fusion protein was purified using GS-4B beads. PreScission protease was applied to remove the GST,the Bradford method used to determine the concentration of purified protein,and SDS-PAGE used to detect its relative molecular weight and purity. The GST pull-down assay was used to study the protein's biological activity. Results The CaME141G protein was successfully purified at a high concentration and purity. The protein could interact with PreIQ protein fragments from the myocardial CaV1. 2 calcium channel C terminal,in a CaME141G concentration-dependent manner. Therefore,CaME141G has the ability to bind with the CaV1. 2 calcium channel. Conclusion This study successfully constructed a CaME141G fusion protein-expressing plasmid and purified the CaME141G protein. This lays a foundation for regulating the function of CaM mutations in the myocardial CaV1. 2 calcium channel,and for the study of its relationship with diseases of the cardiovascular system.
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Aim To construct 3 D structure model of cardiac Cav1.2 channel and check its accuracy and re-liability.Methods Homology model of Cav1.2 chan-nel α1 subunit was constructed using SWISS-MODEL server.The model was submitted to an online testing server built by University of California and scored by it.The binding of Cav1.2 channel with blocker or drug was simulated by MOE software molecular docking pro-gram to check the model′s accuracy and reliability.Re-sults Both the target sequence Cav1.2 α1 C and the template sequence Cav1.1 α1 S searched by SWISS-MODEL server belonged to L-type Ca2+channel.Since the homology was 7 1.5% revealed by sequence align-ment,homology modeling was performed using automa-ted mode.L-type Ca2+ channel blockers Verapamil, Nifedipine and Diltiazem could bind to the 3 D structure model of Cav1.2 channel,while sodium channel bloc-ker TTX could not.Furthermore,active ingredient of traditional Chinese drug Praeruptorin A and Berberine could also bind to the 3D structure model of Cav1.2 channel.Conclusion The 3 D structure model of Cav1.2 channel was constructed successfully,which provides reliable materials for further studies and estab-lishes the foundation for the application of homology modeling in the study of 3 D structure prediction of ion channels.
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Objective To investigate the impact of enhanced recovery after surgery (ERAS) program on postoperative recovery in patients undergoing laparoscopic colorectal resection. Methods Eighty-four patients undergoing laparoscopic colorectal resection from March 201 5 to June 201 6 (55 males,29 females,aged 36-78 years,ASA physical status Ⅰ or Ⅱ),were randomly divid-ed into two groups (n = 38 each).Patients in group E were received epidural block combined with general anesthesia,and a series of perfect ERAS strategies,such as strengthen preoperative educa-tion, maintaining perioperative normothermia, perioperative goal-directed fluid therapy, intraoperative and postoperative analgesia.While the patients in group C received routine anesthetic management.The volume of fluid,the nasopharyngeal temperature,the time of recovery of bouel sound,first anal exhaust,eating fluid food,ambulation and remove of the catheter were recorded in two groups.Furthermore,time of PACU after surgery,the total days of hospitalization and total hos-pital costs were recorded.Results The volume of fluid [(1 328 ± 64)ml vs.(2 463 ± 135 )ml]in group E were significantly lower than group C (P <0.05),the nasopharyngeal temperature [(36.2± 0.2)℃ vs.(35.1±0.5)℃]was significantly higher in group E (P <0.05).Compared with group C,the time of recovery of bowel sound [(33.4 ± 12.5 )h vs.(42.8 ± 14.3 )h],first anal exhaust [(43.6±13.9)h vs.(60.7±1 5.4)h],eating fluid food [(26.8±4.1)h vs.(67.4±13.5)h],first ambulation [(7.4±1.6)h vs.(26.5±3.8)h]and remove of the catheter [(29.2±6.1)h vs.(5 1.8 ±7.6) h ], time of PACU [(26.4 ± 8.5 ) min vs.(37.2 ± 1 1.6 ) min ], the total days of hospitalization [(7.5±0.9)d vs.(9.7±1.2)d]were significantly shorter (P <0.05),and hospital costs [(2.1±0.6)ten thousand yuan vs.(2.6±0.8)ten thousand yuan]were significantly decreased (P <0.05).The incidence of adverse reactions such as nausea and vomiting (2.4% vs.21.4%),pru-ritus (7.1% vs.23.8%),agitation (4.8% vs.26.2%)and chills (0% vs.1 9.0%)were significantly lower in group E (P <0.05).Conclusion ERAS program applied to patients undergoing laparoscopic colorectal resection can reduce the intraoperative sufentanil consumption,avoid the occurrence of postoperative hypothermia, accelerate recovery of gastrointestinal function, which can obviously reduce the hospitalization costs and shorten the hospitalization time.
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Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.
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Objective To explore whether dithiothreitol(DTT)is helpful for PreScission Protease to cut off the GST from GST?CT3 protein. Meth?ods The pGEX?6P?3/CT3 recombinant plasmid was transfected into Escherichia coli BL21,and the GST?CT3 fusion protein was purified by B?PER method. PreScission Protease was applied with 10 mmol/L DTT to cut off the GST,then the SDS?PAGE was performed for identification of the CT3 protein. Results Without DTT,it was very difficult for PreScission Protease to cut off the GST from GST?CT3 protein. However,in the pres?ence of 10 mmol/L DTT,PreScission Protease could cut off the GST easily as identified by SDS?PAGE. Conclusion 10 mmol/L DTT can help Pre?Scission Protease to cut GST from GST?CT3 protein,so as to achieve high concentration of CT3.
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Objective To observe the effects of nonylphenol(NP)on the intracellular calcium concentration changes and cell proliferation,and the involvement of GPR30 receptor in H9c2 cell. Methods The intracellular calcium concentration changes were recorded by using intracellular calcium determination method and cell proliferation was observed by MTT method in H9c2 cell. Results NP(1×10-10 mol/L)increased the intra?cellular calcium concentration changing amplitude and promoted the proliferation of H9c2 cells,while NP(1×10-6 mol/L)decreased intracellular calcium concentration changing amplitude and suppressed cell proliferation. G15 could block the promoting effect of 1×10-10 mol/L NP on the intracel?lular calcium concentration and cell proliferation,but could not block the inhibition of 1×10-6 mol/L NP on the intracellular calcium increase and cell proliferation. Conclusion The results indicate that NP affect rapid calcium signal changes and cell proliferation in non?monotonic dose dependent manner,and its mechanism may be due to the different involvement of GPR30 receptor in different concentrations.
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Objective To explore the expression and significance of HDGF and ADAM9 in esophageal cancer.Methods HDGF and ADAM9 expression in 113 patients with esophageal cancer (78 males and 35 females) with ages ranging from 28 to 87 years,averaged (61.2 ± 8.6) years,were analyzed by SP immunohistoehemical staining.Results The positive rates of HDGF and ADAM9 in esophageal cancer (78.8％ and 68.1％ respectively) were significantly higher than those in normal tissue(all were 30％).ADAM expressions were not correlated with age,sex,tissue types,lymph nodes metastasis,location and tumor size (P ＞ 0.05) ;the positive expression rate of HDGF and ADAM9 was related to differentiation,PT stages,and 5-year survival rate (P ＜0.05).HDGF was correlated with PT stage.Conclusion The expressions of HDGF and ADAM9 in esophageal cancer tissues were more higher than normal esophageal tissues,this means they could promote the development of tumor cells transformation,multiple and moving,and may be an useful tool for providing information about the targeted therapy and prognosis.
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Objective To study the expression and clinical significance of anty-apoptosis protein BAG-1 in non-small cell lung cancer(NSCLC),and its relationship with cell apoptosis and expression of Bcl-2 protein. Methods Immunohisto chemistry streptavidin-peroxidase conjugated (SP)method was used to examine the expression of BAG-1protein and Bcl-2 protein,and terminal deoxynucleotidyl transferase mediated UTP nick end labeling(TUNEL) method was used to examine the apoptosis index in 54 cases of NSCLC.The expression of BAG-1 protein in 20 cases of normal bronchus mucosa tissue also be detected as control. Results Express positive rate of BAG-1 protein in NSCLC is 74.07%,obviously higher than that in normal bronchus mucosa tissue(positive rate is 5%).In cases of NSCLC,the expression of BAG-1 protein has not correlation with the age,gender,pathologic classification,but have closed correlation with lymph node metastasis,degree of differentiation,pTNM stage(P