Research progress of intravital microscopy of the pulmonary immune environment / 中国胸心血管外科临床杂志

@#The development of intravital microscopy (IVM) has enabled researchers to perform in-situ, real-time observations of pulmonary micro-circulation at the cellular level, and has become instrumental for researching the immune micro-environment of pulmonary diseases. This article introduces the developments in constructing the pulmonary imaging window and summarizes the current light microscopy techniques used for lung IVM with regard to its relevant functional and application features, which includes wide field fluorescence microscopy, confocal microscopy, as well as two-photon microscopy. It then provides examples of IVM of pulmonary immune response in inflammation and infection in murine models, and finally specifies the technological limitations to provide reference for researchers to systematically learn and understand the technology.

Characteristics of corneal stromal demarcation line after different cross-linking methods in keratoconus and its influence on the cross-linking effect / 中华实验眼科杂志

Objective:To compare the characteristics of corneal stromal demarcation line after different surgical methods of riboflavin/ultraviolet A corneal collagen cross-linking (CXL) in early keratoconus, and analyze the influence of the demarcation line on the cross-linking effect.Methods:A non-randomized controlled clinical study was conducted.Sixty-nine eyes of 69 patients treated with riboflavin/ultraviolet A CXL in the Eye Hospital of Shandong First Medical University from May 2019 to February 2021 were included.According to the cross-linking methods, the patients were divided into epithelium-on treatment group (21 eyes) and epithelium-off treatment group (48 eyes). There were 25 eyes in 5.4 J energy group and 44 eyes in 7.2 J energy group.The morphology and changes of corneal stromal cross-linking reaction (corneal stromal demarcation line) were observed at 2 weeks, 1, 3 and 4 months after operation.Changes in the thinnest corneal thickness (TCT), uncorrected visual acuity (UCVA, LogMAR), best corrected visual acuity (BCVA, LogMAR) and corneal maximum curvature (Kmax) were recorded.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Eye Hospital of Shandong First Medical University (No.2019.05). Written informed consent was obtained from each subject.Results:Of the 69 eyes after operation, 44 eyes (63.77%) had demarcation lines, and 25 eyes (36.23%) had no demarcation lines.The occurrence rate of demarcation lines in the epithelium-on treatment group was 79.17%(38/48), which was significantly higher than 28.57%(6/21) in the epithelium-off treatment group ( χ2=16.186, P<0.01). The occurrence rate of demarcation line in 5.4 J energy group was 72.00%(18/25), and the 7.2 J energy group was 56.80%(25/44), with no significant difference ( χ2=1.565, P=0.302). Slit lamp microscopy and anterior segment-optical coherence tomography showed that the demarcation line appeared at 1-2 weeks after operation, gradually converged and strengthened after 1 month, turned diffuse, blurred and faded by degrees after 2-3 months, and basically disappeared after 4 months.The depth of the demarcation line reached 141-423 μm, with an average depth of (263.44±84.22)μm.Scanning laser confocal microscopy showed that corneal stromal cells were activated and light reflection was enhanced after CXL.Collagen fibers extended vertically and horizontally, crisscrossed, and were in a reticular arrangement.The TCT decreased from preoperative (458.69±38.28)μm to (443.86±36.54)μm at 4 months after operation, showing a statistically significant difference ( t=6.705, P<0.001). There was no significant difference in the TCT reduction between groups with and without demarcation lines ( t=1.684, P=0.100). At 4 months postoperatively, the UCVA of all eyes increased from preoperative 0.74±0.37 to 0.69±0.38, and the difference was statistically significant ( t=2.109, P=0.039). There was no significant difference in BCVA between before and after operation ( t=1.006, P=0.319). There was no significant difference in change of UCVA and BCVA between groups with and without demarcation lines ( t=0.065, P=0.949; t=0.346, P=0.730). There was no significant difference in Kmax in all patients between before and after operation ( t=0.050, P=0.950). There was no significant difference in the Kmax change between groups with and without demarcation lines ( t=-0.739, P=0.464). The change in TCT in the epithelium-off treatment group was significantly greater than that in the epithelium-on treatment group ( t=2.815, P=0.008). There was no significant difference in UCVA, BCVA and Kmax changes between epithelium-on and epithelium-off treatment groups (all at P>0.05). There was no obvious corneal scarring, infectious keratitis, corneal endothelial decompensation or other complications. Conclusions:The demarcation line after CXL may be a sign of the depth of cross-linking reaction, which is more prone to occur after the epithelium-off operation method.Both the epithelium-on and epithelium-off operation methods have similar therapeutic effects.Demarcation line after different cross-linking methods has no significant influence on the cross-linking effect in keratoconus.

Treatment of rheumatoid arthritis by injection of sinomenine solid lipid nanoparticles under a fluorescence endoscopic laser confocal microscope / 中国中药杂志

A fluorescence endoscopic laser confocal microscope(FELCM) was used to direct the injection of sinomenine solid lipid nanoparticles(Sin-SLN) into the joint, and the in vitro effectiveness of Sin-SLN in the treatment of rheumatoid arthritis(RA) was evaluated. Sin-SLN was prepared with the emulsion evaporation-low temperature curing method. The Sin-SLN prepared under the optimal conditions showed the encapsulation efficiency of 64.79%±3.12%, the drug loading of 3.84%±0.28%, the average particle size of(215.27±4.21) nm, and the Zeta potential of(-32.67±0.84) mV. Moreover, the Sin-SLN demonstrated good stability after sto-rage for 30 days. The rabbit model of RA was established by the subcutaneous injection of ovalbumin and complete Freund's adjuvant. Five groups were designed, including a control group, a model group, a Sin(1.5 mg·kg~(-1)) group, a Sin-SLN(1.5 mg·kg~(-1)) group, and a dexamethasone(positive drug, 1.0 mg·kg~(-1), ig) group. The control group and the model group only received puncture treatment without drug injection. After drug administration, the local skin temperature and knee joint diameter were monitored every day. The knee joint diameter and the local skin temperature were lower in the drug administration groups than in the model group(P<0.05, P<0.01). FELCM recorded the morphological alterations of the cartilage of knee joint. The Sin-SLN group showed compact tissue structure and smooth surface of the cartilage. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the serum le-vels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α). The findings revealed that the Sin-SLN group had lower IL-1 and TNF-α levels than the model group(P<0.05, P<0.01). Hematoxylin-eosin(HE) staining was employed to reveal the pathological changes of the synovial tissue, which were significantly mitigated in the Sin-SLN group. The prepared Sin-SLN had uniform particle size and high stability. Through joint injection administration, a drug reservoir was formed. Sin-SLN effectively alleviate joint swelling and cartilage damage of rabbit, down-regulated the expression of inflammatory cytokines, and inhibited the epithelial proliferation and inflammatory cell infiltration of the synovial tissue, demonstrating the efficacy in treating RA.

Effect of repeated intravitreal Ranibizumab and Aflibercept injections on the corneal nerves in patients with macular edema / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of repeated intravitreal injection of ranibizumab and aflibercept on corneal nerve of patients with macular edema.METHODS: A total of 64 patients(64 eyes)enrolled in our hospital from June 2021 to June 2022 were treated with intravitreal injection of anti-vascular endothelial growth factor(VEGF). There were 20 cases(20 eyes)of diabetic macular edema, 19 cases(19 eyes)of wet age-related macular degeneration and 25 cases(25 eyes)of retinal vein occlusion. Corneal confocal microscope was used to collect images of corneal subbasal nerve plexus before injections and at 1mo after each intravitreal injection based on 3+pro re nata(PRN)treatment regimen. Furthermore, the length and density of corneal nerve were measured.RESULTS: There was no significant difference in corneal nerve density of patients injected with aflibercept between pre-injection and post-injection(P&#x003E;0.05), while the corneal nerve length after 2nd and 3rd injections was lower than that of pre-injection(all P&#x003C;0.01). There were no significant changes in corneal nerve density and length in patients with intravitreal injections of ranibizumab(all P&#x003E;0.05), and there was no significant differences in corneal nerve density and length after 3 injections of the two drugs(all P&#x003E;0.05).CONCLUSION: Repeated intravitreal anti-VEGF drug may affect corneal nerve to some extent. For patients who need repeated intravitreal injections of anti-VEGF, attention should be paid to the changes of corneal nerves.

Clinical and immunopathological study of rejection after bioengineered keratoplasty: a report of 5 cases / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the mechanism of rejection of bioengineered keratoplasty by the laser confocal microscope and immunopathological studies.<p>METHODS: Retrospective case study. Five patients(5 eyes)who underwent bioengineered keratoplasty for infectious keratitis from Sep.2018 to Dec.2020 and were immunologically rejected included. All observers were monocular. Slit-lamp was used to examine corneal transparency and corneal neovascularization. The cornea was dynamically observed under laser confocal microscopy, the density of subepithelial langerhans cells and stromal inflammatory cells was recorded after graft rejection, and the contralateral eye was used as the control group for statistical analysis. The infiltration and distribution of CD4⁺ cells, CD8⁺ cells and inflammatory cells were observed after immunohistochemical staining of bioengineered corneas during corneal allograft.<p>RESULTS: Confocal laser microscopy showed that a large number of langerhans cells were activated under the corneal epithelium of the intraoperative eye, and the activation ratio was significantly different from that of the contralateral eye(χ²=38.29, <i>P</i><0.001). The stromal layer was rich in inflammatory cells, which was significantly different from that of the contralateral eye(<i>t</i>=32.5, <i>P</i><0.05). Immunohistochemical staining and HE staining were performed on the bioengineering corneal tissue. CD4⁺ and CD8⁺ cells were observed in all corneal stromal layers, and only one case had a large number of eosinophils.<p>CONCLUSION: Immunopathology of rejection after bioengineered keratoplasty confirmed that T-cell-mediated cellular immunity plays an important role in the pathogenesis, and hypersensitivity to xenoantigen may be involved in it.

Confocal microscopy observation of corneal nerve plexus and Langerhans cells in patients with systemic diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#Corneal confocal microscope(CCM)is a non-invasive instrument for the study of living cornea and is increasingly being used to evaluate corneal nerve plexus and Langerhans cell lesions. This paper reviews the recent advances in corneal nerve plexus and Langerhans cell lesions from the perspective of the use of CCM in the study of systemic diseases such as diabetes mellitus, Sjogren's syndrome, multiple sclerosis, thyroid-associated ophthalmopathy, fibromyalgia, chronic migraine and Parkinson's disease.

Clinical effect of anterior corneal stroma acupuncture on corneal epithelial cell dysfunction / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the clinical efficacy of anterior stromal puncture(ASP)for the corneal epithelial cells dysfunction(CED).<p>METHODS: Sixteen patients with CED underwent ASP in Wuhan Union Hospital from September 2015 to December 2015 were included. Uncorrected visual acuity, ocular surface disease index(OSDI), corneal fluorescence staining, corneal epithelial thickness, full corneal thickness, corneal subepithelial dendritic cell density, corneal endothelial cell density and corneal epithelial nerve density were observed and recorded at preoperative, 1mo and 3mo postoperatively, respectively.<p>RESULTS: Totally 16 patients compared with preoperatively, there was a significant increase in uncorrected visual acuity and corneal epithelial nerve density(<i>P</i><0.05)or a significant decrease in OSDI, corneal fluorescence staining, corneal epithelial thickness, full corneal thickness and corneal subepithelial dendritic cell density(<i>P</i><0.05)at 1mo postoperatively; while there was no significant difference in corneal endothelial cell density(<i>P</i>>0.05). And compared with 1mo postoperatively, there was a significant decrease in corneal subepithelial dendritic cell density(<i>P</i><0.05)and a significant increase in corneal epithelial nerve density(<i>P</i><0.05)at 3mo postoperatively, while there was no significant difference in uncorrected visual acuity, OSDI, corneal fluorescence staining, corneal epithelial thickness, full corneal thickness and corneal endothelial cell density(<i>P</i>>0.05).<p>CONCLUSION: ASP was effective for CED. Corneal confocal microscopy was essential for the evaluation of ASP efficacy, which can guide the clinical work better and establish the termination of intervention.

Confocal microscopy features in chronic corneal edema / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To report <i>in vivo</i> confocal microscopic(IVCM)in corneal edema.<p>METHODS: Twenty-one patients with clinically diagnosed corneal edema were involved. All cases, in addition to control group of five normal eyes, were examined with IVCM. <p>RESULTS: IVCM findings in corneal edema showed that epithelial bullae appeared as dark, round areas with well-defined margins in all of the patients; Subepithelial fibroblasts and scarring; Other IVCM features included absent(57%)or reduced(43%)subbasal nerves in central corneal; Apart from the associated scar tissue, BZ presented a branching pattern of fine, darklines in all of the patients. Furthermore, fine or coarse granularity with avariable degree of reflectivity were noted in the anterior stroma in 13 of the 21 cases(62%); The number of keratocytes in the stroma of corneas was less than that in the normal corneas measured by using confocal microscopy in all of the patients; Endothelial changes with total obscuration of endothelial cell borders were seen in all of the patients. IVCM findings in the control group showed normal patterns.<p>CONCLUSION: This is the study in which IVCM features of corneal edema have been found in detail. Subepithelial fibroblasts, reduced subbasal corneal nerves and stromal keratocyte were well documented in this study. With increasing popularity of endothelial keratoplasty this work supports the role of IVCM in quantitative evaluation of corneal edema in preoperative stages, as well as after surgery.

Real-time assessments of liver quality in cold ischemia injury based on the technique of laser scanning confocal microscope / 中华器官移植杂志

Objective To assess the quality of donor liver allografts by employing laser scanning confocal microscope (LSCM ) and clinical liver function tests .Methods Sprague-Dawley rats were used for establishing cold ischemia models of liver allografts .According to different timepoints of cold ischemia ,four groups of CIT1h ,CIT6h ,CIT12h and CIT24h were designated .At the end of cold ischemia time (CIT) of each group , perfusion and preservation fluids were collected and fluoresceins perfused . After LSCM examinations ,tissue samples were harvested for HE examination .Finally a comparison was made between LSCM results and hematoxylin & eosin (HE) examinations .Also some relevant clinical parameters were detected in preserving and flushing fluids .Results Both LSCM examination and pathological examination indicated that the quality of liver allografts decreased significantly with the elapsing of time . Only the difference of LDH was statistically significant (P<0 .001) .Conclusions LSCM may be used for evaluating the ex vivo qualities of liver allografts .Simple handling and time efficiency re great advantages of LSCM .As compared with alanine transaminase (ALT ) and aspartate transaminase (AST ) ,LDH is a better indicator reflecting the quality of liver allografts .

La lente dermatológica… historia y algo más / Dermatological vision lens... history and something else

El microscopio es un instrumento, que desde hace muchos años es herramienta diagnóstica en Dermatología. La evolución del microscopio ha ido en sinergia con el avance de la tecnología, siendo el desarrollo de imágenes microscópicas digitalizadas, fuente de estudio en la época actual. El uso de estas imágenes en Dermatología ha permitido realizar diagnósticos en tiempo real; situación que vence la celeridad diagnóstica y aminora tanto para el médico como para el paciente, la lentitud de otras técnicas. El conocimiento sobre los diferentes métodos que se utilizan actualmente, para confirmar diagnósticos de enfermedades cutáneas, como: la dermatoscopía, la dermatoscopía multiespectral y la microscopía confocal de reflectancia, son imprescindibles para la formación del dermatólogo de hoy en día, ya que, con ellos se resumen pasos para el manejo definitivo de los pacientes. Con esta disertación, realizaremos un breve análisis de la historia de la microscopía, desde sus inicios hasta la era digital y los beneficios que se obtienen, con el uso de cada una de estas técnicas diagnósticas.

The microscope, is an instrument that for many years is used as a diagnostic tool in Dermatology. The evolution of the microscope has gone in synergy with the advancement of technology, being the development of digitized microscopic images a source of study and research in the present time. The use of these images in Dermatology has carried out diagnoses in real time; situation that overcomes the diagnostic speed and reduces for both the doctor and the patient the slowness of other techniques. The knowledge about the different methods currently used to confirm skin diseases, such as dermatoscopy, multispectral dermoscopy and confocal reflectance microscopy, is essential for the formation of today's dermatologist, since they summarize the steps for the definitive management of patients. With this dissertation, we will make a brief analysis of the history of microscopy from its beginnings to the digital age, and the benefits obtained with the use of each of these diagnostic techniques.

Application of Laser Scanning Confocal Microscope to Morphologic Research of Acupuncture and Moxibustion / 针刺研究

Laser scanning confocal microscope (LSCM) has been used in the field of morphological research of acupuncture and moxibustion for more than 20 years. It is one of the important tools for revealing the structure of acupoints and the effect of acupuncture from histological perspective. With the help of technical advantages of LSCM, the quality of morphological research of acupuncture and moxibustion has been greatly improved, helping us gain a deeper understanding about the structure of acupoints and meridians as well as histochemical changes induced by acupuncture/moxibustion intervention. In order to promote the application of LSCM in acupuncture and moxibustion, we simply reviewed some recent studies in this field and combined them with our experience, trying to provide some technical suggestions. We expect that the technique of LSCM could be integrated into more experiments in acupuncture medicine to provide more powerful morphological evidence for exploring the underlying mechanisms of acupuncture and moxibution therapies.

Histological changes of cornea in vivo of patients with xerophthalmia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the changes of corneal cytology of patients with different degrees of xerophthalmia of the types of deficient aqueous production and over evaporation with confocal microscopy. METHODS:Retrospective case series was adopted with three - dimensional corneal topography and anterior segment analysis system (Sirius system). A total of 33 typical dry eye patients (total 65 eyes) with deficient aqueous production and 28 typical dry eye patients (total 55 eyes) with over evaporation were selected from optical center of the First Affiliated Hospital of Harbin Medical University from December 2016 to June 2017, which were grading according to dry eye degrees. The corneal epithelial cell (surface epithelial cell, pterygoid epithelial cell, basal epithelial cell), Langerhans cell (dendritic cell),stroma (deep stromal layer and superficial stromal layer), the density and form of endothelial cells were observed and analyzed with the confocal microscope. RESULTS: Confocal microscope showed that the number of corneal epithelial cells decreased with the increasing of dry eye severity in patients with tear deficiency and evaporative. The results were statistically significant (P<0.01). The sizes of each layer of corneal epithelium cells were uneven, and the arrangement was irregular. The diameter of corneal sub-epithelial nerve fiber of deficient aqueous production dry eye became thinner with irregular arrangement. With the increase in severity of dry eye,the degree of curvature and branches gradually increased. The activation degree of Langerhans cells of evaporative dry eye was directly proportional to the severity of dry eye(P<0 01). There was no significant changes in the number of superficial stromal cells in the two types of dry eye (P>0 05). The number of deep stromal cells in the two types of dry eye had no relations with the severity of dry eye (P>0. 05). There was no significant changes in the number of corneal endothelial cells in the two types of dry eye (P>0.05). CONCLUSION:The Sirius system can be used for non-contact and non-invasive examination and classification. And there is changes of the density and morphology in each layers of cornea cells of two types of dry eye with the confocal microscope.

Topical ozone therapy: An innovative solution to patients with herpes zoster / 中南大学学报(医学版)

Objective:To observe the clinical efficacy and safety of topical ozone therapy for patients with herpes zoster by reflectance confocal microscopy (RCM).Methods:A total of 60 patients with herpes zoster were divided into a control group and an ozone treatment group (n=30).In the control group,patients took oral valacyclovir tablets or granules (0.3 g per day,three times a day) and they were subjected to local weak laser irradiation treatment plustopical 2％ mupirocin ointment twice a day.In the ozone group,the treatment is same as the control group except mupirocin ointment was replaced with topical ozone treatment (hydrotherapy every day plus ozonated oil twice a day).The clinical symptoms,discoid cell and adverse reactions were observed and taken records at day 0,3,7 and 14.Statistical analysis was performed to compare the clinical efficacy between the 2 groups.Results:On the seventh day of treatment,the discoid cells of the ozone group disappeared,and the difference between the control group and the ozone group was statistically significant (P＜0.05).The difference of decreased percentage of pain scores at each time point between the 2 groups was statistically significant (P＜0.05).The clinical efficacy was 100％ in the ozone group and 86.7％ in the control group,with significant difference between the 2 groups (P＜0.05).Conclusion:Topical ozone therapy in patients with herpes zoster is helpful in relieving pain,shortening the course as well as improving the clinical efficacy without obvious adverse reactions.It is worth to be popularized.

Retinal nerve fiber layer and corneal nerve loss in early stage of diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the changes of retinal nerve fiber layer(RNFL)and corneal nerve fibers(CNF)in diabetic patients without diabetic retinopathy(DR)retrospectively. <p>METHODS:Forty eyes of 40 patients with type 2 diabetes mellitus, who were found no diabetic retinopathy, were examined by optical coherence tomography(OCT)and <i>in vivo</i> confocal microscope(IVCM). Forty eyes of 40 healthy participants were only scanned using OCT served as RNFL control group, and another forty eyes of 40 healthy participants were only scanned using IVCM served as CNF control group. The thickness of superior, inferior, nasal, temporal and average RNFL to optic disc were measured using OCT. Corneal nerve fibers length(CNFL)and corneal nerve fibers density(CNFD)were measured using IVCM. <p>RESULTS: The diabetic patients showed decreased thickness of inferior RNFL when compared to controls(<i>P</i>=0.003).No significant differences were found in the thickness of superior, nasal, temporal and average RNFL(<i>P</i>>0.05). The diabetic patients were also found to have lower CNFL and CNFD(<i>P</i><0.01). What's more, in diabetes patients, the average RNFL were positively correlated with CNFL and CNFD(<i>r</i>=0.518, <i>P</i><0.01; <i>r</i>=0.484, <i>P</i>=0.002),and the inferior RNFL were also positively correlated with CNFL and CNFD(<i>r</i>=0.607, <i>P</i><0.01; <i>r</i>=0.573, <i>P</i><0.01). <p>CONCLUSION: RNFL and CNF are early damaged in diabetic patients without DR. RNFL decreases in the inferior quadrant of retina. The loss of RNFL positively correlates with that of CNF significantly.

Effect of Electroacupuncture on Intracellular Calcium Ion Concentration and Expression of Calcium/Calmodulin Dependent Protein Kinase Ⅱ in Lumbar Spinal Cord in Rats with Neuropathic Pain / 针刺研究

OBJECTIVE: To observe the changes of intracellular calcium ([Ca2＋]i) concentration and expression of calcium/calmodulin dependent protein kinaseⅡ (CaMKⅡ) in spinal dorsal horn neurons of spared nerve injury (SNI) rats, so as to explore its mechanisms underlying improvement of neuropathic pain. METHODS: One hundred and ten SD rats were randomly divided into 5 groups: sham control, model, EA, AP-5 and L-NAME groups. The sham group underwent only a simple separation of the sciatic nerve but without ligation and abscission. The neuropathic pain model was established by abscission of the right tibial and common peroneal nerve. EA (2 Hz, 1－3 mA) was applied to right "Weizhong" (BL 40) and "Huantiao" (GB 30) for 30 min, once a day for 7 days, starting from day 11 after surgery. For rats of the AP-5 and L-NAME groups, AP-5 (a competitive antagonist for NMDA receptor, 0.7 mg·kg－1·d－1) and L-NAME (a non-selective antagonist for nitric oxide synthase [NOS], 60 mg·kg－1·d－1) were respectively administrated by intraperitoneal injection, once daily for 7 days. The mechanical pain threshold was measured, and the calcium fluorescence intensity (shown by Fluo-3/AM calcium fluorescence indicator) of the superficial layer of the lumbar spinal cord (L 4－L 6) was measured by immunohistochemical staining and the expression of spinal cord (L 4－L 6) CaMK Ⅱ protein was detected by Western blot (WB). RESULTS: After modeling, the mechanical pain threshold was significantly decreased on day 10 and 16 after operation in comparison with the sham operation group and baseline data of pre-operation in each group (P<0.01), and remarkably increased in the EA, AP-5 and L-NAME groups relevant to the model group on day 16 (P<0.01, P<0.05), while the effect of EA was significantly superior to that of AP-5 and L-NAME groups (P<0.05), suggesting a reduction of EA analgesia after administration of AP-5 and L-NAME. The concentration of intracellular [Ca2＋]i was significantly higher in the model group than in the sham group, and considerably lower in the EA, AP-5 and L-NAME groups than in the model group (P<0.01, P<0.05). Moreover, the expression level of CaMKⅡ shown by WB and immunohistochemical staining was significantly higher in the model group than in the sham group (P<0.05) and obviously lower in the EA group (not the AP-5 and L-NAME groups) than in the model group on day 16 after the intervention (P<0.05). It suggests an involvement of glutamate NMDA receptor and NMDAR-NOS/NO signaling in the analgesic effect and CaMKⅡ expression down-regulation of EA. CONCLUSIONS: EA can ease pain in rats with neuropathic pain, which is closely related to its effect in reducing the calcium concentration and the expression of CaMKⅡ in the lumbar spinal cord, possibly mediated by glutamate NMDA receptor and NMDAR-NOS/NO signaling.

Effect of sulbactam sodium/cefoperazone sodium on neutrophil extracellular traps of patients with recurrent lower respiratory tract infection / 吉林大学学报(医学版)

Objective:To detect the neutrophil extracellular traps (NETs) formation in the peripheral blood of the patients with recurrent respiratory tract infection,and to evaluate the effect of sulbactam sodium/cefoperazone sodium on the formation of NETs.Methods:A total of 36 patients with recurrent respiratory tract infection (case group) and 30 healthy volunteers (healthy control group) were selected.The NETs formation of subjects in two groups was detected by confocal microscope and scanning electron microscope (SEM).According to the appearance of neutrophils,the formation of NETs was classified as grade Ⅰ,Ⅱ and Ⅲ,the number of NETs formation cells of subjects in two groups was calculated.The formation of NETs of the patients in case group were detected before and after treated with sulbactam sodium/cefoperazone sodium.Results:The number of NETs formation cells of grade Ⅰ and Ⅱ of the patients in case group was more than that in healthy control group (P<0.05);while the number of NETs formation cells of grade Ⅲ of the patients in case group was less than that in healthy control group (P<0.05).The number of NETs formation cells of grade Ⅰ and Ⅱ of the patients in case group were significantly decreased (P<0.05),while the number of NETs formation cells of grade Ⅲ was significantly increased (P<0.05) after treated with sulbactam sodium/cefoperazone sodium.Conclusion:A lot of NETs with high antibacterial function can be formed in the patients with recurrent lower respiratory tract infection,and sulbactam sodium/cefoperazone sodium can inhibit the formation of NETs.

Effects of chronic alcohol exposure on hippocampus and cerebral cortex neurons in mice / 药学学报

This study was performed to investigate the changes of the number, morphology and ultrastructure of the central nervous system of mice during the long-term alcohol exposure. Mice at 60 days in age were used to establish the long-term alcohol exposure model. The structure of the central nervous system, such as nuclear antigen, dendritic spines and synapses, were labeled by the methods of immunocytochemistry and DiI (1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethy lindocarbocyanine perchlorate) scattering. The results showed that prolonged alcohol exposure could promote apoptosis of nerve cells in the central nervous system, and inhibit the proliferation of neural stem cells, which reduced the number of nerve cells in the central nervous system. Long-term ethanol exposure can also lead to a decrease in the density of dendritic spines of neuron, a smaller number of synapses (connections between nerve cells), and some changes in synaptic ultrastructure. The density of nerve cells and their dendritic spines, as well as the changes of synaptic ultrastructure, suggest that the function of nerve cells may be low.

Protective effects of choline on myocardial ischemic rat heart and its potential mechanisms / 重庆医学

Objective To study the protective effects of choline on myocardial ischemia rat heart and its potential mecha -nisms .Methods Ischemia hypoxia environment was simulated with low value of pH (pH 6 .8) and lack of oxygen .Calcium currents were recorded by whole cell patch under the voltage clamp configuration .The alternations in[Ca2 + ]induced by KCl was detected by laser scanning confocal microscope in ventricular myocytes ,then disccuss the effects of choline on calcium and calcium store in cells . Results The normalized peak currents of ICa-L in ventricular myocytes were larger in pH 6 .8 group than those in pH 7 .4 group , which can be attenuated by choline .The(Ca2 + )i induced by KCl in ventricular myocytes were significantly increased in pH 6 .8 Ty-rode solution and its increasing can be suppressed by choline .4-DAMP can inhibit the suppressing effect of choline .Conclusion The possible mechanism of M 3 receptor involved in the protection of ischemic myocardium by inhibiting myocardial cells in ICa-L ,in-hibiting intracellular calcium overload .

Changes in intracellular Ca2 + in the ureter smooth muscle cells of rats with neuropathic urinary tract dysfunction and its significance / 中华实用儿科临床杂志

Objective To investigate the changes in intracellular Ca2+ in the ureter smooth muscle cells (USMC) of rats with neuropathic urinary tract dysfunction (NUTD) and their significance.Methods Forty-five rats were randomly and averagely divided into NUTD group,experimental control (EC) group and blank control (BC) group.The NUTD group was operated with a spinal cord transection at the first lumbar level and the sacral cord was destroyed;in EC group the spinal process was partly bitten at the same position,but the spinal cord was not transected;BC group was given no operations.One week later,the video-urodynamic was performed to observe the acontractile detrusor (ACD),vesicoureteral reflux (VUR) and urinary tract dysfunction in rats among the NUTD group,EC group and the BC group.Video-urodynamic assessment was performed at the sixth week after operation.Ureter smooth muscle cells (USMC) were obtained by collagenase digestion.Intracellular Ca2 + in the USMC were observed by laser scanning confocal microscope.Then the effects of Bay K8644(10-8 mol/L,10-7 mol/L,10-6 mol/L) on cytosolic Ca2+ concentrations([Ca2+] i) in NUTD group were studied by calculating the fluorescence intensity.Results ACD and no detrusor overactivity were found in all rats in NUTD group and without vesicoureteral reflux.Immunofluorescence method confirmed that the cells were USMC.Compared with BC group (31.44 ± 2.82) and EC group (32.06 ± 3.67),the fluorescence intensity (FI) of intracellular Ca2 + in USMC was much lower in the NUTD group (9.80 ± 1.11),and there was significant difference(P ＜ 0.05).Bay K8644 (10-8 mol/L,10-7 mol/L,10-6 mol/L) increased the FI of [Ca2 +] i in a concentration-dependent manner,which were 3.80 ± 1.30,10.04 ± 2.15,19.89 ± 2.06,respectively,and there was significant difference (P ＜ 0.05).Conclusions The decrease in Ca2 + concentration in the ureter smooth muscle cells may be one of the important factors for the primary ureteral dysfunction of NUTD.And calcium channel agonist can be meaningful for adjusting abnormal Ca2+ concentration in USMC of NUTD.

Vasodilation Effect of Extract of Jasminum samba / 医药导报

Objective To observe the vasodilation effect of extract of Jasminum samba (EJs), a kind of traditional Chinese medicine, on ex vivo rat thoracic aortic rings, and to investigate its mechanism. Methods On ex vivo aortic ring perfusion device, influence of EJs on contraction of the aorta induced by phenylephrine (PE) or potassium chloride (KCl) was observed. Influence of N-nitro-L-arginine-methylester ( L-NAME ), barium chloride ( BaCl2 ), glibenclamide ( Gli ) on vasodilating effect of EJs (0. 5, 1, 2, 4, 8 g·L-1 ) was detected. Effect of EJs on the contraction of calcium chloride (CaCl2 ) and PE in Ca2+-free medium was detected. [ Ca2+ ] i in vascular smooth muscle cells was determined by using laser scanning confocal microscope (LSCM). Results In blood vessels with intact endothelium, EJs concentration-dependently decreased PE- or KCl-induced vasoconstriction, the maximum dilating effect being (105. 0±3. 2)% and (78. 0±6. 5)% , respectively; L-NAME affected the vasodilatory effect of EJs on thoracic aorta rings ( P<0. 01), the maximum dilatory effect being (58. 0 ± 6. 9)% . BaCl2 and Gli had significant influence on vasodilation of EJs, and the contraction was obviously attenuated (P<0. 01), the maximum dilatory effect being (37. 0±5. 2)% and (78. 0±10. 0)% , respectively. EJs significantly inhibited contracting effect of PE on thoracic aorta rings in Ca2+-free medium (P<0. 01). The maximum contraction effect was (70. 0±6. 3)% . EJs inhibited CaCl2-induced vasoconstriction (0. 5-8 mmol·L-1 ), and vasoconstriction was decreased by (65. 0±3. 2)% . LSCM recorded that Fmax / F0 of 4 and 8 g·L-1 EJs was (2. 0±0. 2) and (1. 5±0. 2), respectively. Conclusion EJs exerted a dose-dependent vasodilating effect on rat isolated aorta rings. The mechanism might be related to promoting NO release, activating K+channels and decreasing the concentration of cytoplasmic Ca2+.