Intervention effect of Tuina pressing and kneading the Huantiao (GB30) acupoint on NF-κB p65 protein at spinal cord dorsal horn in sciatica rats / 中国骨伤

OBJECTIVE@#To observe the analgesic effect of Tuina by pressing and kneading the Huantiao (GB30) acupoint on rats with chronic constriction injury (CCI) and to explore the analgesic mechanism of Tuina on sciatica rats.@*METHODS@#Thirty-two SPF male SD rats weighing 180 to 220 g were randomly divided into fore groups:blank group (without any treatment), sham group (only exposed without sciatic nerve ligating), model group (sciatic nerve ligating) and Tuina group (manual intervention after lsciatic nerve ligating). The CCI model was prepared by ligating the right sciatic nerve of the rats, on the third day of modeling, the rats in the Tuina group were given pressing and kneading the Huantiao (GB30) point for 14 days, and the changes of paw withdrawal threshold(PWT), paw withdrawal latency(PWL) were measured before and on the 1st, 3rd, 7th, 10th, 14th and 17th days after modeling. The changes of sciatic functional index(SFI) were measured before and on the 1st and 17th day after modeling. The morphological changes of the sciatic nerve were observed by hematoxylin-eosin(HE) staining;and the differences in NF-κB protein expression in the right dorsal horn of the spinal cord of rats were detected.@*RESULTS@#Following modeling, there was no significant difference in PWT, PWL and SFI between the blank group and the sham group (P>0.05), but the PWT, PWL and SFI of the model group and the Tuina group decreased significantly (P<0.01). After manual intervention, the pain threshold of rats in Tuina group increased. On the 8th day of manual intervention (the 10th day after modeling), PWT in Tuina group increased significantly compared with that in model group (P<0.01). On the 5th day of manual intervention (the 7th day after modeling), the PWL of the massage group was significantly higher than that of the model group (P<0.01). The pain threshold of rats in Tuina group continued to rise with the continuous manipulation intervention. After 14 days of manipulative intervention, the sciatic nerve function index of rats in the Tuina group increased significantly(P<0.01). Compared with the blank group and sham group, the myelinated nerve fibers of sciatic nerve in the model group were disordered and the density of axons and myelin sheath was uneven. Compared with the model group, the nerve fibers of rats in the Tuina group were gradually continuous and the axons and myelin sheath were more uniform than those in the model group. Compared with the blank group and sham group, the expression of NF-κB protein in the right spinal dorsal horn of the model group was significantly increased(P<0.01). Compared with the model group, the expression of NF-κB protein in the right spinal dorsal horn of rats in Tuina group decreased significantly(P<0.01).@*CONCLUSION@#Pressing and kneading the Huantiao (GB30) point restores nerve fiber alignment;and improves the PWT、PWL and SFI in the CCI model by decreasing NF-κB p65 protein expression in the spinal dorsal horn. There fore, Tuina demmstrates an analgesic effect and improves the gait of rats with sciatica.

Experimental research progress of genetically modified mesenchymal stem cells in the treatment of radiation-induced lung injury / 中华放射医学与防护杂志

Radiation-induced lung injury is a common complication of thoracic malignant tumor radiotherapy and severe nuclear accident injury. Currently, there is no effective treatment on this injury. Mesenchymal stem cells (MSCs) are a group of cells with multi-directional differentiation potential and they can protect lung tissue from radiation damage by homing to the injured site and differentiating to the damaged tissues, secreting cytokines and immune regulation. Further, the genetically modifying mesenchymal stem cells have not only the main characteristics of MSCs, but also can efficiently and stably express or knock down a certain of target genes, thereby enhancing or reducing the sensitivity of mesenchymal stem cells to various physiological stimulus and enhancing its therapeutic effect in radiation-induced lung injury, providing new ideas and new strategies for clinical treatment. This paper reviewed the relevant research progress in recent years.

Screening of the key miRNA downstream of TLR2 and validating the function of miR-21 in radioprotection / 中华放射医学与防护杂志

Objective:To screen the key miRNA downstream of TLR2 and explore the function of the miR-21.Methods:Wild type (WT) and TLR2 KO mice were irradiated with 60Co γ-ray to compare their survivals. The downstream miRNAs of TLR2 signaling pathway were screened by RNA sequence in BMCs, and their expressions were verified by QT-PCR. Cell lines with overexpression or knockdown of a miRNA were established to evaluate the function of miRNA. Results:The radiosensitivity of TLR2 KO mice was higher than that of TLR2 WT mice( χ2=4.490, 13.100, 7.928, P<0.05). The bone marrow transplantation experiment proved that the increased radiosensitivity of TLR2 KO mice was related to BMCs ( χ2=4.291, P<0.05). A total of 55 differentially expressed genes were screened by RNA sequence ([log2 Fold Change]>0.95, Q<0.05), of which 28 were up-regulated and 27 were down-regulated. QT-PCR assay determined that miR-21 was down-regulated in BMCs of TLR2 KO ( t=9.420, P<0.01) and MyD88 KO ( t=10.700, P<0.01) mice. It was proved by QT-PCR that the expressions of IL-6 ( t=13.790, P<0.05) and TNF-α ( t=14.280, P<0.05) were increased in a TLR2 dependent manner after PAM3CSK4 stimulation. Overexpression of miR-21 promoted viability of EL4 cells ( t=5.951, P<0.05) and NIH/3T3 cells ( t=4.786, P<0.05) and reduced BMCs apoptosis in WT ( t=4.842, P<0.05) and TLR2 KO ( t=10.520, P<0.05) mice after radiation. Inhibition of miR-21 decreased the viability of EL4 cells ( t=4.815, P<0.05) and NIH/3T3 cells ( t=4.042, P<0.05). Conclusions:miR-21 plays a key regulatory role in the process of TLR2 radioprotection, which may be related to the up-regulation of IL-6 and TNF-α.

Advances in research of HIV transmission networks / 中华医学杂志(英文版)

Transmission network analysis is a crucial evaluation tool aiming to explore the characteristics of the human immunodeficiency virus epidemic, develop evidence-based prevention strategies, and contribute to various areas of human immunodeficiency virus/acquired immunodeficiency syndrome prevention and control. Over recent decades, transmission networks have made tremendous strides in terms of modes, methods, applications, and various other aspects. Transmission network methods, including social, sexual, and molecular transmission networks, have played a pivotal role. Each transmission network research method has its advantages, as well as its limitations. In this study, we established a systematic review of these aforementioned transmission networks with respect to their definitions, applications, limitations, recent progress, and synthetic applications.

Role of 11beta-hydroxysteroid dehydrogenase in bone metabolism / 中国组织工程研究

BACKGROUND: Glucocorticoids play an essential role in osteoblast differentiation, but excessive glucocorticoids will inhibit the osteoblastic phenotype. Regulation of 11β-hydroxysteroid dehydrogenase (11β-HSD) contributes to optimizing the effect of glucocorticoids.OBJECTIVE: To summarize the effect of 11β-HSD in bone metabolism.METHODS: The corresponding author retrieved PubMed and CJFD databases for the articles published before 2016 using the keywords "11β-HSD, bone" in English and Chinese, respectively. Totally 115 articles were retrieved, including 98 English and 17 Chinese articles, and finally 45 eligible articles were included in accordance with the inclusion criteria.RESULTS AND CONCLUSION: Glucocorticoid action can be regulated at prereceptor level by 11β-HSD. 11β-HSD1 activity may predict individual susceptibility to glucocorticoids, which instructs the individualized application of glucocorticoids precisely. 11β-HSD1 activity is closely related to osteoblast differentiation and the presence of an intrinsic differentiation-driven molecular switch inhibits the activity of 11β-HSD1. Instead of regulating the mesenchymal progenitors directly, gucocorticoids regulate the mesenchymal progenitors to differentiate into cranial skeleton mainly through mature osteoblasts. Short-term glucocorticoid exposure directly increases 11β-HSD1 activity and continuous exposure to glucocorticiod indirectly inhibits 11β-HSD1 activity.

Progresses on mechanism of the relationship between gut microbiota and radiation injuries / 中华放射医学与防护杂志

Recently,researchers have paid attention to the relationship between gut microbiota and human health and attempted to investigate the effect of gut microbiota on radiation injury.More and more evidence showed that normal gut microbiota could maintain human health through Toll-like receptors,immune system and inflammatory reaction.Improvements of gut microbiota spectrum and its balance have become an effective strategy for the treatment of certain diseases.This paper reviewed the relationship between gut microbiota and radiation injury and underlying mechanisms,in order to provide novel theoretical evidence and guideline for the therapy of radiation enteritis and other diseases.

Enhancing efficiency of remote mobile NBC decontamination / 医疗卫生装备

Objective To explore and solve the problems of the decontamination group of NBC medical rescue brigade.Methods The problems of NBC medical rescue brigade were discussed from the aspects of decontamination equipment systematization and intellectualization,personnel training and etc.Results Some suggestions were put forward including developing standardized,systematized,intelligent and human-oriented decontamination equipment,strengthening personnel training and etc.Condusion The efficiency of the decontamination group of NBC medical rescue brigade has to be enhanced from the aspects of technology,equipment and personnel.

In vitro assembly of Ebola virus nucleocapsid-like complex expressed in E. coli

Ebola virus (EBOV) harbors an RNA genome encapsidated by nucleoprotein (NP) along with other viral proteins to form a nucleocapsid complex. Previous Cryo-eletron tomography and biochemical studies have shown the helical structure of EBOV nucleocapsid at nanometer resolution and the first 450 amino-acid of NP (NPΔ451-739) alone is capable of forming a helical nucleocapsid-like complex (NLC). However, the structural basis for NP-NP interaction and the dynamic procedure of the nucleocapsid assembly is yet poorly understood. In this work, we, by using an E. coli expression system, captured a series of images of NPΔ451-739 conformers at different stages of NLC assembly by negative-stain electron microscopy, which allowed us to picture the dynamic procedure of EBOV nucleocapsid assembly. Along with further biochemical studies, we showed the assembly of NLC is salt-sensitive, and also established an indispensible role of RNA in this process. We propose the diverse modes of NLC elongation might be the key determinants shaping the plasticity of EBOV virions. Our findings provide a new model for characterizing the self-oligomerization of viral nucleoproteins and studying the dynamic assembly process of viral nucleocapsid in vitro.

Overview of Ebola virus vaccine / 生物工程学报

Ebola virus (EBOV) causes hemorrhagic fever, resulting in mortality rates as high as 90% among infected humans and non-human primates (NHPs). The 2014 Ebola epidemic in West Africa is the severest in history, leading to WHO taking all control measures to stop any possibility of cross-border outbreaks. Because no licensed vaccines or effective therapeutics against EBOV are available, the current outbreak management has been limited to palliative care and barrier methods to prevent transmission. Several promising experimental EBOV vaccines have demonstrated protection in NHPs against lethal EBOV challenge, and some progresses have been made through clinical trials of EBOV vaccine candidates. It is believed there will be some licensed vaccine available in the near future to control EBOV outbreaks. In this review we provide some insights for further development of EBOV vaccines.

Down regulation of miR-203 in radiation-induced thymic lymphoma promoted cells proliferation and inhibited apoptosis / 中华放射医学与防护杂志

Objective To investigate the role of miR-203 in radiation-induced thymic lymphoma (RITL).Methods A 60Co irradiator was used for total-body irradiation.MicroRNAs(miRNAs) level was assayed by qRT-PCR.Cell proliferation was assayed by MTT assay.Cell apoptosis was examined by fluorescence activated cell sorter (FACS).Dual luciferase reporter assay system was used to detect the 3'UTR reporter.Results MiR-203 was down-regulated in RITL tissues.Overexpression of miR-203 strongly inhibited the proliferation of both NIH3T3 cells and EL4 cells and vice versa.MiR-203 inhibited cells proliferation and induced apoptosis via TANK-binding kinase (TBK1),SLUG (SNAI2) and Cyclin D1 (CCND1).Conclusions Radiation down-regulated the level of miR-203 in thymic,which promoted radiation-induced thymic lymphoma by targeting TBK1,SNAI2 and CCND1.

Insights into battles between Mycobacterium tuberculosis and macrophages

As the first line of immune defense for Mycobacterium tuberculosis (Mtb), macrophages also provide a major habitat for Mtb to reside in the host for years. The battles between Mtb and macrophages have been constant since ancient times. Triggered upon Mtb infection, multiple cellular pathways in macrophages are activated to initiate a tailored immune response toward the invading pathogen and regulate the cellular fates of the host as well. Toll-like receptors (TLRs) expressed on macrophages can recognize pathogen-associated-molecular patterns (PAMPs) on Mtb and mediate the production of immune-regulatory cytokines such as tumor necrosis factor (TNF) and type I Interferons (IFNs). In addition, Vitamin D receptor (VDR) and Vitamin D-1-hydroxylase are up-regulated in Mtb-infected macrophages, by which Vitamin D participates in innate immune responses. The signaling pathways that involve TNF, type I IFNs and Vitamin D are inter-connected, which play critical roles in the regulation of necroptosis, apoptosis, and autophagy of the infected macrophages. This review article summarizes current knowledge about the interactions between Mtb and macrophages, focusing on cellular fates of the Mtb-infected macrophages and the regulatory molecules and cellular pathways involved in those processes.

Minimally invasive treatment for fresh acromioclavicular dislocation and the distal clavicle fracture / 中国骨伤

<p><b>OBJECTIVE</b>To explore the minimally invasive treatment for fresh acromioclavicular dislocation and the distal clavicle fracture.</p><p><b>METHODS</b>Thirty skeletons of human shoulder were measured and compared, and the normal data on healthy people were measured with the help of ultrasound-guided. So the invasion point was located at the cross between subclavian axis and the line from coracoid tip to apophysis behind cone ligament node. From January 2001 to January 2010, 127 patients with fresh acromioclavicular dislocation and distal clavicle fracture were treated with minimally invasive internal fixation after locating the invasive point at the body surface. Among the patients, 97 patients were male and 30 patients were female, ranging in age from 19 to 56 years, with an average of 43 years. According to Rockwood classification, among 93 patients with fresh acromioclavicular dislocation, 67 patients were type III, 11 patients were type IV and 15 patients were type V. All the 34 patients with distal clavicle fractures were associated with coracoclavicular ligament broken. The duration from injury to operation ranged from 1 to 8 days. The therapeutic effects were evaluated by using the of shoulder scoring system, University of California (UCLA).</p><p><b>RESULTS</b>After the minimally invasive treatment, all the patients had completely reduction at early time. One hundred and thirteen patients were followed up,and the duration ranged from 13 to 15 months,averaged 14 months. Nine patients had screw loose slightly within 30 days, but the reductions and functions were acceptable. Seven patients had complications of frozen shoulder and recovered in 6 months. The average UCLA shoulder score was (32.0 +/- 4.7), and 87 patients got an excellent result, 20 good and 6 fair.</p><p><b>CONCLUSION</b>This minimally invasive treatment has advantages such as little trauma and low cost, which is worthy of clinical applications.</p>

Expression and crystallographic studies of a fungal immunomodulatory protein LZ-8 from a medicinal fungus Ganoderma lucidum / 生物工程学报

LZ-8 protein, isolated from a well known Chinese traditional medicinal fungus Ganoderma lucidum, is the first member of fungal immunomodulatory protein, members of which have been isolated from a variety of medicinal and edible mushrooms in the last two decades. The protein plays a multifunctional and important role in modulating immune system. In this report, in order to get LZ-8 protein crystals, the LZ-8 gene was expressed and purified by affinity chromatography, gel filtration chromatography and anion exchange chromatography subsequently. The protein was then crystallized using the hanging-drop vapour-diffusion method. The LZ-8 crystals were obtained and the phase information was calculated by X-ray diffraction. The resolution of LZ-8 crystals is 3.2A. This study will provide an insight into the structure of fungal immunomodulatory proteins.

Suppression subtraction hybridization in screening of differentially expressed genes in radiation-induced malignant transformation cellular model / 第二军医大学学报

Objective: To screen for the differentially expressed genes during irradiation-induced malignant transformation of human bronchus epithelium cells (BEAS-2B). Methods: Suppression subtraction hybridization (SSH) was used to construct a subtracted cDNA library of differentially expressed genes during irradiation-induced malignant transformation of BEAS-2B cells. Then the subtracted library was screened by PCR and the differential fragments were sequenced and analyzed with BLAST. Fluorescent real-time quantitative PCR was used to investigate some of the differentially expressed genes. The new EST was registered in GenBank. Results: Then 40 clones were chosen to be sequenced from the library of increased expression and decreased expression respectively according to the length of insertion element. Totally 73 sequences were obtained from the 80 sequenced clones. Forty-one sequences were decreased in the transformed cells; BLAST analysis indicated that there were 6 known sequences, 20 unknown sequences, 7 void sequences and 8 repeated sequences. Thirty-two sequences were increased in the transformed cells; Blast analysis indicated that there were 14 known sequences, 9 unknown sequences, and 9 repeated sequences. Fluorescent quantitative PCR revealed that, compared with control group, the expression of MY06, HACE1, ZNF143, and HNRPH1 were significantly increased in the radiation transforming group, with their mRNAs increased by 3.49, 29.38, 12.99 and 5.00 folds, respectively. Compared with control group, the expression of PCBP2, RPL15, and TCERG1 in the radiation transforming group was significantly decreased, with their mRNAs decreased by 1.89,48.77 and 11.95 folds, respectively. The 29 unknown sequences were registered in the GenBank (ID: EB643220-EB643248). Conclusion: The cDNA library has been successfully established for malignant transformation cellular model by suppression subtractive hybridization; the library includes a number of unknown genes. The increased gene ZNF143 is associated with cell proliferation and cell division. TCERG1, as an assistant transcription activation factor, plays an important role in the mRNA transcription and later modification. PCBP2, a Polyc connection protein, plays a modulating role in protein translation. These genes have not been reported in the radiation carcinogenicity.

Protective effects of proanthocyanidin on radiation-induced injury of human-derived cells / 第二军医大学学报

Objective: To investigate the protective effects of proanthocyanidin on radiation-induced injury of AHH-1 and HIEC cells, and to explore the possible molecular mechanism. Methods: CCK-8 assay was applied to examine the survival of AHH-1, HIEC cells after γ-ray irradiation with or without proanthocyanidin pretreatment. Annexin-V/PI staining and flow cytometry analysis were used to evaluate the apoptosis and cell cycle of AHH-1 and HIEC cells. The expression of Bcl-2 protein was analyzed by Western blotting assay. Results: Pretreatment with proanthocyanidin significantly increased the cell survival rate after radiation(P<0.01). The apoptosis rates of cells in the proanthocyanidin+radiation group were (11.78% at 4 Gy[AHH-1] and 5.32% at 8 Gy[HIEC], respectively) greatly lower than those of the single radiation group (26.38% at 4 Gy[AHH-1] and 12.45% at 8 Gy[HIEC], respectively, P<0.01). Moreover, the inhibition of Bcl-2 protein expression in AHH-1, HIEC cells was attenuated in the proanthocyanidin + radiation group. Conclusion: Proanthocyanidin has satisfactory protective effect on radiation-induced cellular injury; the mechanism may be related to the attenuation of Bcl-2 protein inhibition.

Differential radiobiological effects of carboxyfullerene C_3 on K562 and AHH-1 cells / 中华放射医学与防护杂志

Objective To investigate the application prospective of carboxyfullerene C_3 as a radioprotectant or assistant for tumor radiotherapy.Methods Different concentrations of C_3 were incubated with K562 and AHH-1 cell,CCK-8 assay and trypan blue rejection test were performed to examine the influence of C_3 on the cell viability.Annexin V/PI staining and flow cytometry assay were applied to assess the cell cycle and apoptosis after 7-ray irradiation.Results C_3 showed little toxicity to AHH-1 cell with the survival rate over 95% ,but 600 mg/L of C_3 markedly inhibited the growth of K562 cell (82%) .Pretreatment of 100 mg/L C_3 significantly increased the survival rate of AHH-1 cell after 4 Gy irradiation compared with the single radiation group(71.3% vs 90.3%) ,but decreased the apoptosis rate (26.3% vs 12.6%) ,while the survival rate of K562 cell was decreased and the apoptosis rate was elevated with the increase of C_3 concentration.Moreover,the cell cycle analysis revealed the G_2 phase block in AHH-1 cell after radiation exposure was mitigated by C_3 pretreatment,but that in K562 cell was aggravated.Conclusions C_3 has good radioprotective effects on AHH-1 cells.For K562 cell,C_3 could inhibit the cell proliferation,promote the radiation induced apoptosis and aggravate the G_2 phase block.

Studies of modifying effects of fullerene on ~(60)Co - γ irradiated zebrafish / 中华放射医学与防护杂志

Objective To evaluate the modification of C_(60) on the radiation effects of ~(60)Co-γ irradiation on zebrafish.Methods The adult and embryonic zebrafish were used as model organisms to examine the potential of C_(60) to elicit oxidative stress responses on the surviving rate,hatching rate and malformation occurrence,both upon exposure to light or in the dark.Reactive oxygen species (ROS) production and DNA damage were examined as the possible underlying mechanism.Results 500 × 10~(-9) nano-C_(60) waterborne exposure could enhance the γ-irradiation effects by decreasing adult fish survival upon light exposure,which resulted in ROS and DNA damage increasing.The hatching rates were also inhibited with higher malformation,though dark exposure did not make any enhancement,except that the 5000× 10~(-9) C_(60) would inhibit larvae hatching and induced more malformation.Conclusions Waterborne nano-C_(60) exposure may enhance the radiation effects on zebrafish,ROS production and DNA damage increasing may be the underlying mechanism.

Nephrotic syndrome complicated with intracranial venous thrombosis treated with urokinase: report of 5 cases / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the effect of urokinase and low molecular weight heparin in children with nephrotic syndrome complicated with intracranial venous thrombosis.</p><p><b>METHODS</b>Urokinase and low molecular weight heparin were administered to the 5 patients intravenously. The initial dose of urokinase was 2000 - 4000 U/(kg.d), the initial pulse dose was 20 000 - 40 000 U given within 15 - 30 minutes, and the left was infused by using a pump, from the second day 2000 U/(kg.d) urokinase was infused daily for 3 to 7 days. During the treatment thrombin time (TT), activated partial thromboplastin time (APTT) were tested 3 times every week, with particular attention to bleeding. Low molecular weight heparin 100 - 120 AXaIU/kg, 1 or 2 times per day was hypodermally injected for a course of two weeks. Anti-platelet drugs: long-term oral administration of dipyridamole 3 - 5 mg/(kg.d) was applied 2 - 3 times every day for 3 months.</p><p><b>RESULTS</b>The clinical symptoms disappeared after one month of the combined therapy of urokinase, low molecular weight heparin and dipyridamole in 5 cases of nephrotic syndrome complicated with intracranial venous thrombosis in children, the plasma viscosity returned to normal in 1 month, activated partial thromboplastin time, prothrombin time, fibrinogen degradation products returned to normal in 1 to 2 months, venous thrombosis disappeared after 1 to 3 months in head CT or MRI examination, showing the cerebral venous sinus thrombosis complete recanalization without relapse cases in follow-up.</p><p><b>CONCLUSION</b>The early application of urokinase and low molecular heparin and anti-platelet coagulation drugs was effective. The early diagnosis, treatment and prevention of intracranial vein thrombosis in patients with nephrotic syndrome is important.</p>

An increase in intracelluar free calcium ions modulated by cholinergic receptors in rat facial nucleus / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Ca(2+) in the central nervous system plays important roles in brain physiology, including neuronal survival and regeneration in rats with injured facial motoneurons. The present research was to study the modulations of intracellular free Ca(2+) concentrations by cholinergic receptors in rat facial nucleus, and the mechanisms of the modulations.</p><p><b>METHODS</b>The fluorescence intensity of facial nucleus in Fluo-3 AM loaded acute brainstem slices was detected by applying intracellular free Ca(2+) measurement technique via confocal laser scanning microscope. The changes of fluorescence intensity of facial nucleus indicate the average changes of intracellular free Ca(2+) levels of the neurons.</p><p><b>RESULTS</b>Acetylcholine was effective at increasing the fluorescence intensity of facial nucleus. Muscarine chloride induced a marked increase of fluorescence intensity in a concentration dependent fashion. The enhancement of fluorescence intensity by muscarine chloride was significantly reduced by thapsigargin (depletor of intracellular Ca(2+) store; P < 0.01), rather than Ca(2+) free artifical cerebrospinal fluid or EGTA (free Ca(2+) chelator; P > 0.05). And the increase of fluorescence intensity was also significantly inhibited by pirenzepine (M(1) subtype selective antagonist; P < 0.01) and 4-DAMP (M(3) subtype selective antagonist; P < 0.01). In addition, fluorescence intensity was markedly increased by nicotine. The enhancement of fluorescence intensity by nicotine was significantly reduced by EGTA, nifedipine (L-type voltage-gated Ca(2+) channel blocker), dihydro-beta-erythroidine (alpha4beta2 subtype selective antagonist), and in Ca(2+) free artificial cerebrospinal fluid (P < 0.01), but not in the presence of mibefradil (M-type voltage-gated Ca(2+) channel blocker) or thapsigargin (P > 0.05).</p><p><b>CONCLUSIONS</b>The data provide the evidence that muscarinic receptors may induce the increase of intracellular free Ca(2+) levels through the Ca(2+) release of intracellular Ca(2+) stores, in a manner related to M(1) and M(3) subtypes of muscarinic receptors in rat facial nucleus. Nicotine may increase intracellular free Ca(2+) concentrations via the influx of extracellular Ca(2+)+ mainly across L-type voltage-gated Ca(2+) channels, in a manner related to the alpha4beta2 subtype of nicotinic receptors.</p>

Study of in vitro endotoxin neutralization effect and antibacterial activity of limulus anti-lipopolysaccharide factor simulated peptide REMP2 / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the role of REMP2 derived from limulus anti-lipopolysaccharide factor in neutralizing endotoxin in vitro and its antibacterial activity.</p><p><b>METHODS</b>(1) REMP2 and PMB in the concentrations of 100.00, 10.00, 1.00, 0.10, 0.01 micromol/L were respectively mixed with LPS (lEU/mL), with PMB as positive control. The LPS concentrations in different specimens were determined by routine method, and the neutralizing percentage was respectively calculated. (2) After adding isotonic saline (NS), the final concentrations of REMP2 and PMB were 10, 20, 40, 80 micromol/L, and the concentration of LPS was 100 microg/L. The murine monocytic macrophages were stimulated with LPS, then cultured with REMP2 and PMB, with NS in culture as negative control. The content of tumor necrosis factor (TNF)-alpha was determined by ELISA kit. (3) The morphologic changes of Escherichia coli. was observed under electron microscope at 10, 20 and 40 minutes after addition of REMP2 to Escherichia coli suspension (with terminal concentration of REMP2 at 40 micromol/L).</p><p><b>RESULTS</b>There were no significant difference in endotoxin-neutralizing percentages between PMB and REMP2 in concentrations of 0.10, 10.00, 100.00 micromol/L (P > 0.05). The contents of TNF-alpha were 1175 +/- 162, 859 +/- 122, 645 +/- 142, 489 +/- 102 ng/L, respectively,after treatment of 10, 20, 40, 80 micromol/L REMP2, which were obviously lower than that of NS (3463 +/- 218 ng/L, P < 0.01). Under transmission electron microscope, the outer and interior membranes of Escherichia coli were obscure and rough, bacterial bodies were swollen with vacuoles in cytoplasm after treatment with REMP2.</p><p><b>CONCLUSION</b>REMP2 has ability of neutralizing endotoxin and also antibacterial activity.</p>