Objective To investigate the mechanisms of the inhibitory effects of peripheral electrical stimu- lation(PES)on chronic central pain(CCP)after spinal cord injury(SCI).Methods Twenty-four male Sprague- Dawley rats with CCP following SCI were randomly divided into three groups:a group without stainless steel needles implanted (NSSN group,n=8),a group with a stainless steel needle implanted but no peripheral electrical stimula- tion applied(NPES group,n=8)and a PES group(PES group,n=8).The rats' CCP was evaluated through ob- serving their response to nociceptive stimulation by means of the paw withdrawal pressure threshold(PWPT)and the paw withdrawal latency(PWL).Spontaneous pain behaviors including autophagia and scratching were observed at the same time.PES was applied via stainless steel needles inserted into standard acupoints on the hind limps and the back.The expression of the NMDA receptor 1(NR-1)subunit in the spinal cord horn was measured using immuno- chemical methods.Results Compared with the NSSN and NPES groups,CCP in the PES group was alleviated, PWPT and PWL were dramatically increased(P＜0.01)and the expression of NR-1 was obviously decreased (P＜0.01).Conclusion Peripheral electrical stimulation may alleviate chronic central pain after spinal cord injury in rats.

Objective There is still a concern that epidural labor analgesia could affect uterinecontraction.The purpose of this study was to investigate the effect of epidural labor analgesia on uterinecontraction.Methods Forty ASA Ⅰ-Ⅱ primiparous women aged 20-30 yr at full term in normal uncomplicateddelivery were enrolled in this study.They were taller than 1.5 m and weighed less than 100 kg.The amnioticmembrane was artificially ruptured at 3 cm cervical dilation and a catheter was inserted into uterine cavity beyondthe head of the fetus and connected to a maternal-fetal monitor.The patients were randomly divided into 2 groupswith 20 patients in each group:Ⅰ control group received no analgesia and Ⅱ epidural group received continuousepidural analgesia(PCEA).An epidural catheter was placed at L_2-3.After a loading dose of 8-10 ml of the PCEAsolution(0.1% ropivacaine+1 ?g?ml~(-1) fentanyl)PCEA was started(bolus 3 ml,lockout interval 15 min andback ground infusion 6-8 ml?h~(-1)).The height of block was controlled below T_10.Blood samples were taken frommaternal vein at 3 cm cervical dilation(T_1),1h later(T_2)and at delivery(T_3)and from umbilical vein andamniotic fluid was aiso collected for determination of cortisol,PGE_2 and pitocin levels.VAS scores,intrauterinepressure,the frequency and duration of uterine contraction,the use of pitocin(%),incidence of cesareansection,the length of labor and neonatal Apgar scores were recorded.Results The maternal blood eortisolconcentration was significantly lower during PCEA(T_2,T_3)in group Ⅱ than in control group(P

Objective To investigate the effect of dexamethasone on the pulmonary gas exchange in patients undergoing cardiac valve replacement under cardiopulmonary bypass(CPB).Methods Forty ASAⅡorⅢpatients aged 29-47 yrs weighing 50-69 kg undergoing cardiac valve replacement under CPB were randomly divided into 2 groups(n=20 each):dexamethasone group received dexamethasone 0.5 mg?kg~(-1) after induction of anesthesia and control group received normal saline(NS).Blood samples were taken before operation(T_0) immediately before CPB(T_1)and immediately after discontinuation of CPB(T_2)for determination of plasma total and active matrix metallo-proteinase-9(MMP-9)concentration(by enzyme-linked immuno-absorbent and fluorometric enzyme-linked immuno-absorbent assay respectively)and MMP-9 gene expression(RT-PCR).Blood samples were taken from radial artery at T_1 and T_2 for blood gas analysis.A-aDO_2 was calculated.Results MMP-9 gene expression and plasma total and active MMP-9 concentrations were significantly increased at T_2 as compared with those at T_0 in both groups and were significantly lower in dexamethasone group than in control group(P＜0.05 or 0.01).The A-aDO_2 at T_2 was significantly smaller in dexamethasone group than in control group.Conclusion Dexamethasone can inhibit the increase in gene expression,protein synthesis and activation of MMP-9 and decrease in gas exchange across alveolar-capillary membrane caused by CPB and protect the lungs during open heart surgery performed under CPB.

Mechanical ventilation (MV) with large tidal volumes can increase lung alveolar permeability and initiate inflammatory responses,resulting in ventilator-induced lung injury (VILI).The mechanisms of the injurious effects of MV and the genetic susceptibility remain unclear.VILI-related genes such as cysteine-rich angiogenic inducer 61 (Cyr61)have been demonstrated to play a detrimental role in the aggressive ventilation strategies.In the present study,we investigated the involvement of Cyr61 in the VIM and the underlying mechanism.A549 cells were exposed to cyclic stretch of varying durations and then the mRNA and protein levels of Cyr61 were measured by real-time PCR and Western blotting,respectively.Additionally,after exposure ofA549 cells to cyclic stretch for 5 min to 1 h,the expression levels of nuclear factor kappaB (NF-κB) and IL-8 were detected by ELISA and Western blotting.Thereafter,Cyr61 expression was depressed in A549 cells with the siRNA pGenesill.1-Cyr61-3 before the cyclic stretch,and IL-8 secretion and the activation of NF-κB pathways were probed by ELISA and Western blotting,respectively.Moreover,a NF-κB inhibitor (PDTC) and an activator (TNF) were used before mechanical stretch.Realtime PCR and ELISA were performed to detect the mRNA and protein of IL-8,respectively.The results showed that the mechanical cyclic stretch led to increased Cyr61 expression at mRNA and protein levels in A549 cells.Additionally,cyclic stretch also mobilized NF-κB from the cytoplasm to the nucleus and increased IL-8 secretion in A549 cells.The inhibition of Cyr61 blocked the NF-κB activation and IL-8 secretion in response to cyclic stretch.Inhibition of NF-κB attenuated the mRNA and protein expression of IL-8 in A549 cells transfected with Cyr61 siRNA.It was suggested that Cyr61/NF-κB signaling pathway mediates the upregulation of IL-8 in response to cyclic stretch in A594 cells.These findings support the hypothesis that Cyr61 plays a critical role in acute lung inflammation triggered by mechanical strain.

BACKGROUNDTreponema pallidum (T. pallidum) subsp. pallidum is the causative agent of syphilis. Analysis of recombinant antigens of T. pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis. Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies, but the results were controversial. In this research, the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed.

METHODST. pallidum subsp. pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted. The Tp0965 gene was amplified by polymerase chain reaction (PCR). Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system. The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay. The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay.

RESULTSRecombinant protein Tp0965 was expressed successfully in vitro. Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages. Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965, and lack of reactivity of Tp0965 to all 28 uninfected sera. A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization.

CONCLUSIONSThe recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages. The results also demonstrate a potential application for the serodiagnosis of syphilis.

Animals ; Antigens, Bacterial ; genetics ; immunology ; Bacterial Proteins ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Membrane Proteins ; genetics ; immunology ; Polymerase Chain Reaction ; Rabbits ; Syphilis ; immunology ; microbiology ; Treponema pallidum ; immunology ; metabolism

Background Treponema pallidum (T.pallidum) subsp.pallidum is the causative agent of syphilis.Analysis of recombinant antigens of T.pallidum led to the identification of potential candidate antigens for vaccine development and syphilis serodiagnosis.Tp0965 was predicted to be a membrane fusion protein and was found to be reactive with infected human sera in previous studies,but the results were controversial.In this research,the antigenicity and immunoreactivity of recombinant protein Tp0965 were assessed.Methods T.pallidum subsp.pallidum (Nichols strain) was propagated and isolated and the genomic DNA was extracted.The Tp0965 gene was amplified by polymerase chain reaction (PCR).Then the recombinant protein Tp0965 was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid (Ni-NTA) purification system.The reactivities of protein Tp0965 were examined by immunoblot analysis and indirect enzyme-linked immunosorbent assay.The antisera against protein Tp0965 were obtained by immune rabbits and the immunogenicity of antisera were detected by indirect enzyme-linked immunosorbent assay.Results Recombinant protein Tp0965 was expressed successfully in vitro.Immunoblot assay showed that the recombinant protein Tp0965 could be recognized by human syphilitic sera of all stages.Indirect enzyme-linked immunosorbent assay showed there were only 4 of 74 human syphilitic sera that failed to show reactivity to recombinant antigen Tp0965,and lack of reactivity of Tp0965 to all 28 uninfected sera.A low titer of antiserum against Tp0965 in immune rabbits could be detected after the third time of immunization.Conclusions The recombinant antigen Tp0965 shows excellent sensitivity for the reactivity with sera from syphilitic individuals at all stages.The results also demonstrate a potential application for the serodiagnosis of syphilis.

Carotenoids have a range of diverse biological functions and actions, especially playing an important role in human health with provitamin A activity, anti-cancer activity, enhancing immune ability and so on. Human body can't synthesis carotenoids by itself and must absorb them from outside. However, carotenoid contents in many plant are very low, and many kinds of carotenoid are difficult to produce by chemical ways. With the elucidation of carotenoid biosynthetic pathway and cloning genes of relative enzymes from microorganisms and higher plants, it is possible to regulate carotenoid biosynthesis via genetic engineering. This article reviews gene cloning of carotenoid biosynthetic enzymes in microorganisms and higher plants, and advances in the studies of carotenoid production in heterologous microorganisms and crop plants using gene-manipulated carotenoid biosynthesis.
Candida albicans ; genetics ; Carotenoids ; biosynthesis ; Cloning, Molecular ; Escherichia coli ; genetics ; Genetic Engineering ; methods ; Plants ; genetics ; Saccharomyces cerevisiae ; genetics

The possible role of minocycline in microglial activation and neuronal death after cardiac arrest (CA) and cardiopulmonary resuscitation (CPR) in mice was investigated in this study. The mice were given potassium chloride to stop the heart beating for 8 min to achieve CA, and they were subsequently resuscitated with epinephrine and chest compressions. Forty adult C57BL/6 male mice were divided into 4 groups (n=10 each): sham-operated group, CA/CPR group, CA/CPR+minocycline group, and CA/CPR+vehicle group. Animals in the latter two groups were intraperitoneally injected with minocycline (50 mg/kg) or vehicle (normal saline) 30 min after recovery of spontaneous circulation (ROSC). Twenty-four h after CA/CPR, the brains were removed for histological evaluation of the hippocampus. Microglial activation was evaluated by detecting the expression of ionized calcium-binding adapter molecule-1 (Iba1) by immunohistochemistry. Neuronal death was analyzed by hematoxylin and eosin (H&E) staining and the levels of tumor necrosis factor-alpha (TNF-α) in the hippocampus were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that the neuronal death was aggravated, most microglia were activated and TNF-α levels were enhanced in the hippocampus CA1 region of mice subjected to CA/CPR as compared with those in the sham-operated group (P<0.05). Administration with minocycline 30 min after ROSC could significantly decrease the microglial response, TNF-α levels and neuronal death (P<0.05). It was concluded that early administration with minocycline has a strong therapeutic potential for CA/CPR-induced brain injury.
Animals ; Cardiopulmonary Resuscitation ; Cell Death ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Heart Arrest ; pathology ; Hippocampus ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Microglia ; cytology ; drug effects ; Minocycline ; pharmacology ; Neurons ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Coal Mining ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Lung ; chemistry ; pathology ; Male ; Middle Aged ; Occupational Diseases ; metabolism ; Pneumoconiosis ; metabolism ; Tumor Suppressor Proteins ; analysis

Background: Early diagnosis and accurate staging are important to improve the cure rate and prognosis for pancreatic cancer.This study was performed to develop an automatic and accurate imaging processing technique system,allowing this system to read computed tomography(CT)images correctly and make diagnosis of pancreatic cancer faster.Methods: The establishment of the artificial intelligence(AI)system for pancreatic cancer diagnosis based on sequential contrast-enhanced CT images were composed of two processes: training and verification.During training process,our study used all 4385 CT images from 238 pancreatic cancer patients in the database as the training data set.Additionally,we used VGG16,which was pre-trained in ImageNet and contained 13 convolutional layers and three fully connected layers,to initialize the feature extraction network.In the verification experiment,we used sequential clinical CT images from 238 pancreatic cancer patients as our experimental data and input these data into the faster region-based convolution network(Faster R-CNN)model that had completed training.Totally,1699 images from 100 pancreatic cancer patients were included for clinical verification.Results: A total of 338 patients with pancreatic cancer were included in the study.The clinical characteristics(sex,age,tumor location,differentiation grade,and tumor-node-metastasis stage)between the two training and verification groups were insignificant.The mean average precision was 0.7664,indicating a good training effect of the Faster R-CNN.Sequential contrast-enhanced CT images of 100 pancreatic cancer patients were used for clinical verification.The area under the receiver operating characteristic curve calculated according to the trapezoidal rule was 0.9632.It took approximately 0.2 s for the Faster R-CNN AI to automatically process one CT image,which is much faster than the time required for diagnosis by an imaging specialist.Conclusions: Faster R-CNN AI is an effective and objective method with high accuracy for the diagnosis of pancreatic cancer.