RESUMEN
Background@#Recent studies have reported that costoclavicular blocks (CCBs) can consistently block almost all branches of the brachial plexus while sparing the phrenic nerve and provide effective analgesia after shoulder surgery. We aimed to compare the efficacy of the CCB with that of the interscalene block (ISB) as the sole blocking technique for shoulder surgery. @*Methods@#A total of 212 patients undergoing elective arthroscopic shoulder surgery were randomized to receive an ISB or CCB based on a non-inferiority design. All patients received titration sedation with propofol under monitored anesthesia during surgery. The primary outcomes were the proportion of patients with complete motor blockade of the suprascapular nerve (SSN) and incidence of hemidiaphragmatic paralysis (HDP). The secondary outcomes included block-related variables, complications, and postoperative pain scores. @*Results@#The proportion of patients with complete motor blockade of the SSN at 20 min between the CCB and ISB groups (53% vs. 66%) exceeded the predefined non-inferiority margin of −5%, but was comparable at 30 min (87% vs. 91%). The CCB resulted in a significantly lower incidence of HDP (7.55% vs. 92.45%), Horner’s syndrome (0% vs. 18.87%), and dyspnea (0% vs. 10.38%) than the ISB. None of the patients experienced failed blocks or required conversion to general anesthesia. Pain scores were comparable between the groups. @*Conclusions@#Ultrasound-guided CCBs may be comparable to ISBs, with fewer unfavorable complications in patients with impaired lung function undergoing arthroscopic shoulder surgery.
RESUMEN
Objective:To evaluate the feasibility and applicability of using phospholipid-hybridization method for preparing biomimetic microbubbles (Bio-MBs) ultrasound contrast agents.Methods:Leukocyte biomimetic microbubbles (MB leu), platelet biomimetic microbubbles (MB pla) and erythrocyte biomimetic microbubbles (MB ery) were prepared by multiple steps: film-hydration, phospholipid-hybridization, mechanical oscillation. The size and zeta potential of Bio-MBs were measured by dynamic light scattering. A laser scanning confocal microscopy experiment was performed to confirm the presence of membrane proteins on the shell of Bio-MBs. The fluorescence of FITC-labeled typical membrane protein was evaluated using a flow cytometer. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to characterize the membrane protein. Biosafety of Bio-MBs was evaluated by CCK-8 counting kit, blood and major organs. The contrast enhancement effect and stability were observed in vitro and in vivo. An in vivo fluorescence imaging system was performed to evaluate the distribution of Bio-MBs. The application value of biomimetic microbubbles was measured by ultrasound molecular imaging by using ischemia-reperfusion rat models and acute hepatitis rat models. Results:Bio-MBs with spherical shape distributed homogenously, without obvious aggregation. The membrane proteins were successfully integrated into the shell of Bio-MBs.The diameter of three Bio-MBs was similar to that of control microbubbles (MB con) ( P>0.05), three Bio-MBs had a lower zeta potential than MB con ( P<0.05). The Bio-MBs had an appreciable performance in vitro and in vivo biosafety. The Bio-MBs retained the main proteins inherited from cell membrane. Contrast enhanced ultrasound imaging in vitro and in vivo showed that the Bio-MBs had a stable imaging ability.MB leu and MB pla have good targeted imaging effect in two disease models. Conclusions:A series of Bio-MBs ultrasound contrast agents, which have high stability, biosafety and targeted imaging efficiency, were successfully prepared by using phospholipid-hybridization method. This fabrication method for obtaining Bio-MBs can be applied to different clinical scenarios with different cell types in the future.
RESUMEN
Objective:To explore the treatment effect of gefitinib on epidermal growth factor receptor (EGFR)-positive advanced non-small cell lung cancer (NSCLC).Methods:Sixty patients with EGFR-positive advanced NSCLC who were admitted to the 904th Hospital of Joint Logistics Support Force of Chinese PLA from March 2016 to January 2020 were selected. They were divided into gefitinib treatment group (30 cases, treated with gefitinib) and combined treatment group (30 cases, treated with pemetrexed combined with cisplatin) by random number table. The anti-tumor efficacy, levels of tumor markers [serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment antigen (CYFRA21-1) and neuron-specific enolase (NSE)] before and after treatment, adverse reactions and 6-month overall survival (OS) rate were compared between the two groups.Results:The clinical control rate of gefitinib treatment group was higher than that of combined treatment group [76.7% (23/30) vs. 50.0% (15/30), χ2 = 4.593, P = 0.032]. There was no significant difference in the levels of CEA, CYFRA21-1 and NSE between the two groups before treatment (all P > 0.05). The levels of CEA, CYFRA21-1 and NSE after treatment in gefitinib treatment group were (902±41) μg/L, (3.1±0.4) ng/ml and (17.7±2.3) ng/ml. The levels of CEA, CYFRA21-1 and NSE after treatment in combined treatment group were (999±51) μg/L, (4.0±0.5) ng/ml and (19.4±3.1) ng/ml. The levels of CEA, CYFRA21-1 and NSE after treatment in gefitinib treatment group were lower than those in combined treatment group ( t = 7.441, P < 0.01; t = 7.459, P < 0.01; t = 2.486, P = 0.016).The levels of CEA, CYFRA21-1 and NSE after treatment in the two groups were all lower than those before treatment, and the differences were statistically significant (all P < 0.05). There was no significant difference in the incidence of rash, thrombocytopenia, digestive tract reaction, and proteinuria between the two groups [26.7% (8/30) vs. 23.3% (7/30), χ2 = 0.089, P = 0.766; 16.7% (5/30) vs. 13.3% (4/30), χ2 = 0.131, P = 0.718); 30.0% (9/30) vs. 26.7% (8/30), χ2 = 0.082, P = 0.774; 10.0% (3/30) vs. 13.3% (4/30), χ2 = 0.162, P = 0.688]. After 6 months of treatment, the OS rate in gefitinib treatment group was 93.3%, and that in combined treatment group was 83.3%, and there was no statistical difference between the two groups ( χ2 = 1.456, χ2 = 0.228). Conclusion:Gefitinib treatment for EGFR-positive advanced NSCLC patients can enhance the anti-tumor efficacy, reduce the content of tumor markers, and has good safety.
RESUMEN
Objective To evaluate the role of α2A adrenergic receptor (α2AAR) in dexmedetomidine-induced inhibition of TLR4/NF-κB signaling pathway activation during hypoxia-reoxygenation (H/R)caused injury to alveolar type Ⅱ epithelial cells.Methods Type Ⅱ] alveolar epithelial cells of rats RLE6TN cells cultured in vitro were divided into 4 groups (n =6 each) using a random number table method:control group (group C),H/R injury group (group H/R),dexmedetomidine group (group D) and α2A AR small interfering RNA (siRNA) plus dexmedetomidine group (group α2AAR-siRNA+D).H/R was produced by exposing cells to 1％ O2-5％ CO2-94％ N2 for 24 h followed by 4-h reoxygenation.Cells were incubated for 1 h with dexmedetomidine at the final concentration of 1 nmol/L,and then H/R model was established in group D.In group α2AAR-siRNA+D,cells were transfected with 50 nmol/L α2AAR-siRNA,48 h later dexmedetomidine at the final concentration of 1 nmol/L was added,cells were incubated for 1 h,and then H/R model was established.The cell viability was measured using CCK-8 method,cell apoptosis rate was determined by flow cytometry,and the expression of TLR4 and NF-κB was detected by immunofluorescence.Results Compared with group C,the cell viability was significantly decreased,the apoptosis rate was increased,and the expression of TLR4 and NF-κB was up-regulated in group H/R (P＜0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Compared with group H/R,the cell viability was significantly increased,the apoptosis rate was decreased,and the expression of TLR4 and NF-κB was down-regulated in group D (P＜0.05),and no significant change was found in the parameters mentioned above in group α2AAR-siRNA+D (P＞0.05).Compared with group D,the cell viability was significantly decreased,the apoptosis rate was increased,and the expression of TLR4 and NF-κB was up-regulated in group α2AAR-siRNA+D (P＜0.05).Conclusion The mechanism by which dexmedetomidine inhibits TLR4/NF-κB signaling pathway activation may be related to activating α2AAR during H/R-caused injury to alveolar type Ⅱ epithelial cells.
RESUMEN
Objective To explore the feasibility and safety of laparoscopic right hemihepatectomy (LRH) via anterior approach for larger tumors in the right lobe of the liver.Methods A retrospective study was conducted based on the clinical data of ten consecutive patients with large right liver cancer undergoing LRH through anterior approach and thirty-seven patients undergoing open hemihepatectomy by anterior approach in recent 6 years.Results Between the two groups there were no significant difference in gender,average age,the mean tumor size,preoperative liver reserve function,cut margin and intraoperative blood transfusion.The LRH group had less average intraoperative blood loss [(408 ± 158)ml vs.(520 ± 153)ml,t =2.047,P =0.046] and shorter postoperative hospital stay [(11.5 ±2.8)d vs.(16.2 ±4.6) d,t=3.091,P=0.003],longer operation time [(302 ±38)min vs.(251±55)min,t=2.732,P=0.009].There was no perioperative death and no significant difference in complications (20.0％ vs.35.1％,x2 =0.812,P =0.367) and similar median survival time (36 mon vs.29 mon,x2 =1.266,P =0.261).Conclusions LRH via anterior approach for larger tumors in the right lobe of the liver is safe and feasible.
RESUMEN
Objective To explore the feasibility and safety of total laparoscopic radical resection for a patient of Bismuth type Ⅲ a hilar cholangiocarcinoma.Methods This patient underwent right hemihepatectomy combined caudate lobectomy,radical regional lymphadenectomy and Roux-en-Y hepaticojejunostomy under total laparoscopic techniques.Preoperatively the volume of future liver remnant estimated by CT scan was 46％,and indocyanine green retention rate at 15 min (ICG R15) was 6.0％.Results The total laparoscopic surgery was carried out successfully with operation time of 540 min and intraoperative blood loss 300 ml,without blood transfusion.The results of pathological examination showed well-differentiated adenocarcinoma of hilar bile duct with negative tumor margins and no regional lymph node metastasis(0/13).The postoperative recovery was uneventful with hospital stay time of 10 days and without any complications.Conclusion At experienced hands,total laparoscopic radical resection of Bismuth type Ⅲ a hilar cholangiocarcinoma is feasible and safe for selected patients.
RESUMEN
OBJECTIVE:To investigate clinical efficacy and safety of different dosages of bromocriptine in the treatment pro-lactinoma,and its effects on serum prolactin(PRL)and tumor volume. METHODS:A total of 60 patients with prolactinoma were selected from our hospital during Jan.-Dec. 2015 as research objects,and then divided into group A and B according to random number table,with 30 cases in each group. Both groups were given Bromocriptine mesilate tablets orally during meal. Group A was given medicine with initial dose of 2.5 mg/d,increasing to 3.75 mg/d 3 d later,increasing by 2.5 mg every week after 2-3 d,and then recovering to 3.75 mg/d till serum PRL level had been controlled. Group B was given medicine with initial dose of 1.25 mg/d, increasing to 2.5 mg/d 3 d later,increasing by 1.25-2.5 mg every week after 2-3 d,and then recovering to 2.5 mg/d till serum PRL level recovered to normal. Both groups were treated for consecutive 3 months. Clinical efficacies as well as serum level of PRL and tumor size were observed in 2 groups,and the occurrence of ADR was recorded. RESULTS:The total response rate of group A (83.33％) was higher than that of group B (66.67％),without statistical significance (P>0.05). Before treatment,there was no statistical significance in serum level of PRL and tumor size between 2 groups (P>0.05). After 1,2 months of treatment,serum levels of PRL in 2 groups were decreased significantly,and the group A was significantly lower than the group B,with statistical significance(P0.05). After treatment,tumor size of 2 groups were decreased significantly,and large adenoma and giant adenoma size in group A were significantly smaller than group B,with statisti-cal significance(P0.05). The inci-dence of ADR in group A(12 cases,40.00％)was significantly higher than group B(5 cases,16.67％),with statistical signifi-cance(P<0.05). CONCLUSIONS:Increasing dosages of bromocriptine no significant influence on therapeutic effect of prolactino-ma,but it can shorten the time of serum PRL level back to normal,and reduce the tumor size. The incidence of adverse reactions in-crease with the dosage.
RESUMEN
Objective This study aims to investigate the effect of Lipoxin A4 receptor on acute lung injury (ALI) induced by intestine ischemia-reperfusion (IIR). Methods Thirty-two 8-week old SD rats were randomly divided into four groups: sham, intestine ischemia-reperfusion (IIR), IIR + BML111 (BML-111), Boc-2 + IIR +BML111 (Boc-2). BML-111 (1 mg/kg) was given intraperitoneally at the onset of reperfusion in the BML-111 and the Boc-2 group. Boc-2 (50 μg/kg) was given intraperitoneally after anesthesia in the Boc-2 group. Rats were subjected to superior mesenteric artery occlusion consisting of 45-min ischemia and 6-h reperfusion, and the sham laparotomy was served as controls. The lung pathology was assayed by the H&E staining. Lung water content was detected using dry/wet ratio. Concentrations of TNF-α, IL-1β, and IL-6 in lung tissue were determined by ELISA. The protein expression of p38 MAPK and NF-κB of lung was assayed by western blot. Results IIR induced serious ALI, with poor lung pathology and increased lung water content, elevation of TNF-α, IL-1β, and IL-6 levels in lung, accompanied with activation of p38 MAPK/NF-κB pathway. However, BML-111 could inhibit the activation of p38 MAPK/NF-κB pathway, leading to the reductions of TNF-α, IL-1β, and IL-6 in lung and attenuation of IIR-induced ALI. Conclusion BML-111 treatment could attenuate inflammation in lung after IIR injury via inactivating the p38 MAPK/NF-κB signaling pathway.
RESUMEN
Objective To explore the pathological changes of lung, expression of the relevant inflammatory factors and oxidative stress markers of Sprague-Dawley (SD) rats undergoing autologous orthotopic liver transplantation (AOLT). Methods Thirty SD rats were randomized into sham group and AOLT group. The pathological changes of lung, expression of the relevant inflammatory mediators and oxidative stress markers were detected . Results ( 1 ) Compared with the sham group , the pathological scores of lung tissue in AOLT group increased significantly and reached its peak at 8 h after surgery. Then the pathological scores decreased to the level of sham within 24 h to 48 h after surgery; (2)The relative expression of inflammatory mediators including TNF-α, IL-1β, IL-6 and IL-8 increased significantly and reached its peak at 8 h after surgery in AOLT group. Then decreased to the level of sham group within 24 h to 48 h after surgery; (3)The change trends of MDA and H2O2 were similar to inflammatory mediators.The relative SOD expression decreased significantly and touched the nadir at 8h after surgery and then increased. Conclusion The pathological changes of lung expression, the relevant inflammatory mediators and oxidative stress markers of rats underwent AOLT were consistent.
RESUMEN
Objective To investigate the protective effects of astragalus preconditioning on the tolerance of ischemia time of mouse small intestine . Methods C57BL/6 mice were randomly divided into 5 groups (n = 7): sham operation group (Sham group),intestinal ischemia reperfusion group (IR group) and astragalus preconditioning group (ASIR group). IR group and ASIR group include 2 sub-groups respectively, specifically, 2 h reperfusion was performed 45 min (ASIR1) and 60 min (ASIR1) after blocking superior mesenteric artery. Intestinal terminal morphology was observed by light microscope after HE coloration . Serum levels of LPS , DAO and intestinal mucosa TNF-α were measured by ELISA. Intestinal Cyto C expression were detected by immunofluorescence. Results Astragalus preconditioning reduces Chiu′s score significantly. Expression of Cyto C was significantly down-regulated in astragalus preconditioning groups, and levels of LPS, DAO and TNF-αsignificantly decreased. The damages in IR2 group is obviously severe than in IR1, but there were no significant differences between this two groups after pretreatment with astragalus. Conclusion Astragalus preconditioning has obvious protective effects to intestinal ischemia reperfusion, and enhances the tolerance to longer time of ischemia.
RESUMEN
Objective To investigate the effects of dipeptidyl peptidase 4(DPP-4) inhibitor on serum adiponectin (APN) and high sensitivity C-reactive protein (hs-CRP) in patients with type 2 diabetes mellitus.Methods 45 cases were type 2 diabetes were collected from department of endocrinology,wuxi second People's hospital of jiangsu province from January 2015 to April 2015.45 patients with type 2 diabetes mellitus were randomly divided into treatment group (n=23) and control group (n =22) , treatment group was treated with metformin combined with DPP-4 inhibitor and control group was treated without DPP-4 inhibitor.Before and after treatment,fasting plsma glucose(FPG),postprandial 2h glucose(2hPG),adiponectin, hs-CRP and homeostasis model assessment for insulin resistance(HOMA-IR) were measured.Results Adiponectin was significantly higher in treatment group than before(P<0.05),FPG,2hPG,hs-CRP and HOMA-IR were significantly lower than before(P<0.05).Adiponectin in treatment group was significantly higher than control group post-treatment(P<0.05).2hPG,hs-CRP and HOMA-IR in treatment group were siginificantly lower than control group post-treatment (P<0.05).Conclusion DPP-4 inhibitor could improve insulin resistance in type 2 diabetes mellitus by increasing serum adiponectin and decreasing serum Hs-CRP and HOMA-IR.
RESUMEN
We wished to assess the role of chlamydia micro virus capsid protein Vp3 in recombinant molecules, chart its molecular evolution, screen the wild-type strain, and reveal its value in clinical research. Using a protein BLAST multiple-alignment program, we compared various strains of Chlamydia micro virus capsid protein Vp3 sequences. Using a "distance tree" of those results, we created a phylogenetic tree. We applied the Karplus-Schulz method of flexible-region analyses for highly conserved alignments of amino-acid sequences. Gamier-Robson and Chou-Fasman methods were employed to analyze two-level structures of sequences. The Emini method was used for analyses of the accessibility of surface epitopes. Studies of hydrophilic proteins were undertaken using Kyte-Doolittle and Hopp-Woods methods. Analyses of antigen epitopes helped to reveal the antigen index using the Jameson-Wolf method. All sequences in the six strains of chlamydia micro virus capsid protein Vp3 were highly conserved, with the main differences being between Vp3 protein in Chp1 and the other five strains of the micro virus. The viral strain of Vp3 protein was based mainly on micro-alpha helix structures, and multiple epitopes were noted in highly conserved regions. Vp3 protein was highly conserved structurally, and was an important protein of the chlamydiaphage capsid. Vp3 protein has a complicated molecular structure, highly conserved regions with strong immunogenicity, and has considerable research value.
Asunto(s)
Secuencia de Aminoácidos , Proteínas de la Cápside , Química , Genética , Alergia e Inmunología , Chlamydia , Genética , Alergia e Inmunología , Secuencia Conservada , Mapeo Epitopo , Evolución Molecular , Datos de Secuencia Molecular , Recombinación GenéticaRESUMEN
Objective To observe the efifcacy of rosuvastatin in patients with unstable angina and its impact on the levels of lipids, high sensitive C-reactive protein (hs-CRP), homocysteine(Hcy) and troponin I (cTnI). Method 384 patients with unstable angina, from January 2010 to December 2012, were randomly divided into observation group and control group, each group had 192 cases, the control group received simvastatin, the observation group were gave rosuvastatin. The efficacy, and the levels of lipids, hs-CRP, Hcy and cTnI were observed after treatment. Results The total effective rate was 92.19%in observation group which was signiifcantly better than 81.25%in control group (χ2=9.044, P<0.01). Before treatment, the levels of TG , TC, LDL-C, HDL-C, Hcy, hs-CRP and cTnI showed no signiifcant difference, after treatment the levels of TG , TC and LDL-C, Hcy, hs-CRP and cTnI were signiifcantly lower than those before treatment (P<0.01), while, the levels of HDL-C signiifcantly increased than those before treatment (P<0.01), the reducing or increasing levels in observation group were more signiifcant compared with the control group (P<0.01). Conclusion Rosuvastatin treatment in unstable angina not only can reduce plasma lipid, but also reduce their inflammation, and stabilize the arterial plaque for unstable angina, it play an important role in development and prognosis.
RESUMEN
Objective To evaluate the effect of chlamydiaphage virus protein 2(Vp2) on the recombinant virus and virus screening research, and it clinical value thereof. Methods To compare the Vp2 protein sequences to get the conserva-tive region with COBALT. A phylogenetic tree was built with ProteinBlast of Distance tree. The amino acid sequence in the high conservative region was predicted by the methods of Gamier-Robson and Chou-Fasman, and its flexibe regions were predicted by Karplus method. The hydrophilicity plot was predicted by Kyte-Doolittle and Hopp-Woods method. The sur-face probability was analysed by Emini, and the antigenic index was analysed by Jameson-Wolf method. Results The six Chlamydiaphage Vp2 proteins were the highly conserved sequences. There were obvious differences between Chp1Vp2 and other 5 Vp2 proteins. There were the main structure-alpha helix and some cell epitopes in the high conserved region. Con-clusion Vp2 protein is the important component of chlamydia phage capsid with the conservative nature. Vp2 protein has complicated structures and high conservative region with strong immunogenicity, playing a practical value of research in vi-rus recombinantment and screening the wild strains of chlaymdia trachomatis phage.
RESUMEN
[Objective] To observe the specific immune responses induced by the recombinant major outer membrane protein (rMOMP) vaccine against Chlamydia trachomatis E serotype in rhesus monkeys.[Methods] Six rhesus monkeys were equally divided into three groups:adjuvant and protein group vaccinated with purified rMOMP and Freund's adjuvants,adjuvant group immunized with Freund's adjuvants only,and control group immunized with phosphate buffer.All the rhesus monkeys were intramuscularly immunized in the triceps brachii for 3 times at a 2-week interval.Two weeks after the last vaccination,serum,vaginal wash and venous blood samples were collected from the rhesus monkeys,and lymphocytes were isolated from the blood samples.Enzyme linked immunosorbent assay (ELISA) was performed to determine the specific IgG antibody and interferon level in sera and secretory IgA (sIgA) level in wash samples,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferation of lymphocytes after stimulation with Chlaraydia trachomatis serotype E elementary bodies.Delayed hypersensitivity was observed in rhesus monkeys challenged by inactivated Chlamydia trachomatis serotype E elementary bodies.In vitro antibody neutralization assay was conducted with the serum from rhesus monkeys.Indirect immunofluorescenee was used to detect Chlamydia trachomatis in exfoliative vaginal cells from rhesus monkeys from week 1 to 10 after challenge with Chlamydia trachomatis.Data were statistically analyzed by using one-way analysis of variance and least significant difference (LSD) test with the SPSS 14.0 software.[Results] The adjuvant and protein group differed statistically from the adjuvant group and control group in the serum level of specific IgG antibody (1.718 ± 0.213 vs.0.841 ± 0.315 and 0.791 ±0.437,both P＜ 0.05),interferon ((1086 ± 121.730) ng/L vs.(409 + 53.440) ng/L and (162 ± 48.046) ng/L,both P＜ 0.05),lymphocyte proliferation index (7.012 ± 1.026 vs.4.473 ± 1.850 and 1A26 ± 1.104,both P＜0.01 ) and the diameter of nodus in delayed hypersensitivity assay ( ( 1 1 ± 2.134) mm vs.(3 ± 0.914) mm and 0,both P ＜ 0.01 ).After attack,the exfoliative cells kept positive for Chlamydia trachomatis in the adjuvant and protein group from week 1 to 5,and in the other 2 groups from week 1 to 10,but were negative in the adjuvant and protein group from week 6 to 10.[Conclusion] The rMOMP vaccine can induce a specific,protective,humoral and cellular immune response against Chlamydia tracbomatis in rhesus monkeys.
RESUMEN
Objective To detect Chlamydia trachomatis phage Vp1 gene in clinical swab specimens and anti-Vp1 antibodies in serum specimens.MethodsCervical and urethral swab as well as serum specimens were collected from attendees to the sexually transmitted disease(STD) clinic in the Tianjin Institute of STD,during March 2008 to March 2011.PCR was conducted to detect chlamydial phage Vp1 gene in swab samples,enzyme linked immunosorbent assay(ELISA) and Western blot to detect anti-Vp1 antibody in sera.The swab specimens positive for Vp1 gene were subjected to cell culture followed by the detection of Vp1 protein with an immunofluorescence-based method.ResultsTotally,36 out of 1542 swab specimens turned out to be positive for Vp1 gene,and 23 out of 453 serum specimens for anti-Vp1 antibody.No positive results were obtained in the Vp1 gene-positive swab specimens by cell culture and immunofluorescence-based assay.ConclusionThe Vp1 gene of Chlamydial trachomatis phage and anti-Vp1 antibody are successfully detected from clinical swab and serum specimens respectively.
RESUMEN
ObjectiveTo detect the changes of heat shock protein 60 (HSP60) during the subcultures of standard and clinical strains of Chlamydia trachomatis(Ct),and to determine if the absence of chlamydial inclusions is associated with the persistent infection of Ct.MethodsA total of 40 Ct strains isolated by cell culture from patients were included in this study and classified into 2 groups according to whether inclusions appeared after initial culture.Reverse transcription PCR was conducted to quantify the levels of HSP60 in specimens containing Ct after 1-4 subcuhures.Chi-squared test was performed to analyze the relationship between the levels of HSP60 and treatment outcomes of patients.ResultsThe levels of HSP60 in clinical specimens containing Ct serovar E were significantly higher in subcultures prior to the appearance of inclusions than in subcultures with the appearance of inclusions(P ＜ 0.05).The ratio of HSP60:16S rRNA mRNA expression after the first,second,third,fourth passage was 0.38 ± 0.06,0.39 ± 0.03,0.38 ± 0.04 and 0.39 ±0.03 respectively in 18 specimens with inclusions appearing after the initial culture,1.18 ± 0.10,0.28 ± 0.06,0.30 ± 0.03 and 0.29 ± 0.05 respectively in 12 specimens with inclusions appearing after the second culture,1.20 ± 0.04,1.20 ± 0.04,0.28 ± 0.04 and 0.28 ± 0.05 in 10 specimens with inclusions appearing after the third culture.Whether inclusions appeared after the initial culture was associated with the treatment outcome of patients.Inclusions were undetected after the initial culture in 16 of 20 specimens from patients with poor response to treatment,but observed in 14 of 20 specimens from patients who tested negative for Ct after one course of treatment.Conclusions It is implicated that no inclusions form after the initial culture in 80％ of specimens from patients experiencing treatment failure.The Ct strains whose inclusions do not form after passages may be in a persistent state,and the expression of HSP60 is high in these strains.Specimens should be subjected to at least 3 blind passages to avoid missed diagnosis of Ct infection.
RESUMEN
Objective To optimize the concentration of polyethylene glycol (PEG) for the growth of Chlamydia trachomatis (Ct) reference strains D-UW-5/Cx and E-UW-5/Cx,and to evaluate the effects of PEG on the sensitivity of Ct to 4 common antibiotics.Methods After the inoculation of Ct standard strains (D-UW-5/Cx and E-UW-5/Cx) into McCoy cell monolayer,different concentrations of PEG were added into the culture medium followed by centrifugation.After 2 hours of incubation at 37 ℃,the inoculum was removed and a complete culture medium containing 1 μg/mL cycloheximide was added followed by another 48-hour culture.Subsequently,the culture was fixed and subjected to iodine staining for the calculation of Ct inclusions and optimization of PEG for the growth of Ct.Some Ct standard strains were used to infect McCoy cells,with PEG (0.7％,wt/vol) added to the culture medium after inoculation and before centrifugation process,and when the infection rate reached higher than 90％,a microdilution method was utilized to evaluate the minimal inhibitory concentration (MIC) of 4 antibiotics,including azithromycin,minocyline,moxifloxacin and doxycycline.Thirty-one clinical specimens,which had been confirmed to be positive for Ct serovar D or E strain,were inoculated into McCoy cell monolayer for the passage of Ct with or without the presence of 0.7％ PEG.Results The optimal concentration of PEG was 0.7％ for the growth of Ct,and this concentration of PEG could increase the number of inclusion bodies of Ct serovar E by 3.44 folds,and that of Ct serovar D by 3.56 folds.In vitro,the MICs of the 4 antibiotics were consistent between PEG-treated and untreated Ct reference strains.Moreover,PEG notably increased the quantity of inclusion bodies of Ct serovar E or D from clinical specimens after passages.Conclusions PEG (0.7％) can enhance the growth of Ct serovar D and E,but has no obvious influence on antimicrobial susceptibility of Ct.
RESUMEN
Objective To investigate the binding protein of chlamydiaphage phiCPG1 capsid protein Vp1 on chlamydia trachomatis outer membrane.Methods The bacterium with recombinant plasmid Vp1/pet30a( + ) was induced.The expressed protein was purified by gel recycling.FarWestern blot was utilized to' investigate the binding protein of Vp1 on chlamydial outer membrane,including recombinant polymorphic outer membrane protein (rPmp) and major outer membrane protein (MOMP).Results The recombinant protein Vp1 was successfully expressed in E.coli.Monoclonal antibody against Vp1 was used as primary antibody in Western blot,and no specific band was present,which indicated that the monoclonal antibody did not specifically bind with any rPmp.Far-Western blot results showed that there was an obvious band for the rPmpI,but no specific band for other rPmp and MOMP,which suggested that Vp1 could specifically bind with rPmpI protein on the chlamydial outer membrane of serotype D.Conclusions There is a binding site of Vp1 on the chlamydia trachomatis outer membrane.Vp1 may play an important role in the interaction between the chlamydiaphage and the chlamydiae.
RESUMEN
<p><b>OBJECTIVE</b>To make a qualitative analysis on chemical components in Ligustrum lucidum by UPLC-ESI-Q-TOF-MS.</p><p><b>METHOD</b>ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 microm) column was adopted, with methyl cyanides-0.1% formic acid as the mobile phase for gradient elution; ESI ion source was used for mass spectra, and data were collected in positive and negative mode.</p><p><b>RESULT</b>Fourteen compounds of L. lucidum were identified by analyzing positive and negative ion mass spectra information and element composition and comparing controls with data from relevant literature.</p><p><b>CONCLUSION</b>After the separation by ultra high performance liquid chromatography, relative molecular mass was determined by mass spectra, and chemical compounds in L. lucidum were determined by information search through relevant literature data, in order to provide powerful therapeutic material basis for L. lucidum.</p>