Decreased Expression of Urethral Caveolin-1, -2, and -3 in the Rat Model of Overactive Bladder: Potential Mediator of Functional Interaction of Urethra and Urinary Bladder / 대한배뇨장애요실금학회지

Purpose@#To investigate the effect of detrusor overactivity (DO) on the urethral expression of caveolin (CAV)-1, -2, and -3 of urethra in an animal model of cyclophosphamide (CYP)-induced cystitis rat. @*Methods@#Female Sprague-Dawley rats were divided into the control group (n=20) and the cystitis group (n=20). Cystitis was induced by intraperitoneal injection of CYP (200 mg/kg). An urodynamic study was done 3 days after the CYP injection to measure functional change of the urinary bladder and urethra. Cellular localization and expression of CAV-1, -2, and -3 in the rat urethra were determined by immunohistochemistry (IHC) and Western blot. @*Results@#Urodynamic experiments demonstrated a decreased contraction interval in the cystitis group compared to the control (3.9±1.0 minutes vs. 6.6±1.2 minutes, P<0.05). Conversely, contraction pressure increased significantly in the cystitis group compared to the control (22.4±0.7 mmHg vs. 11.5±0.4 mmHg, P<0.05). The urethral pressure was decreased in the cystitis group compared to the control (4.05 ±2.5 mmHg vs. 5.8 ±2.8 mmHg, P <0.05). The IHC and Western blot data showed that CAV-1, -2, and -3 expression decreased significantly in the cystitis group compared control group (P<0.05). @*Conclusions@#The decreased urethral CAV-1, -2, and -3 in the DO rats suggests that CAVs might be related with the functional change of urethra in association with DO of urinay bladder.

Effects and related mechanisms of histone deacetylase inhibitors on the proliferation of choroidal melanoma C918 cell line / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To elucidate the effect of histone deacetylase(HDAC)inhibitor suberoylanilide hydroxamic acid(SAHA)on the proliferation of choroidal melanoma(CM)cell line C918 and to explore the related mechanism.METHODS: Inverted fluorescence microscope was used to observe the effect of different concentrations of SAHA(0.625, 1.25 or 2.5 μmol/L)on the morphology of C918 cell. The cell viability was detected by cholecystokinin octapeptide(CCK-8)assay. Plate clone formation assay and EdU staining were carried out to measure the effect of SAHA on the cell proliferation. Meanwhile, the expressions of cell proliferation-related proteins including c-Myc, CyclinA2 and CDK2, and histone deacetylase 7(HDAC7)and fibroblast growth factor 18(FGF18)were detected by Western blot.RESULTS: Compared with the control group, the cell density was reduced in SAHA. SAHA could also promote cell shrinkage, and the inhibition on cell was in a concentration-dependent manner. CCK-8 assay showed that SAHA treatment decreased cell viability in a dose-dependent manner and the inhibition rate was 80% when SAHA at 2.5 μmol/L. Compared with the control group, Western blot showed that SAHA could suppress the expression of cell proliferation proteins including c-Myc, CyclinA2 and CDK2 in a dose-dependent manner. In addition, 1.25 μmol/L SAHA significantly decreased the numbers of EdU staining positive cells and cell clones. More importantly, SAHA could dose-dependently decrease the expression of HDAC7 and FGF18 compared with control group.CONCLUSION: SAHA could inhibit the proliferation of CM cell line C918 by inhibiting the HDAC7/FGF18 signaling pathway.

Toxicity Management and Efficacy Evaluation of BCMA-CART in the Treatment of Relapsed and Refractory Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the toxicity management and efficacy evaluation of BCMA-chimeric antigen receptor T cells(CART) in the treatment of relapsed and refractory multiple myeloma (MM).@*METHODS@#The efficacy and adverse reactions of 21 patients with MM who received BCMA-CART treatment at the First Affiliated Hospital of Wenzhou Medical University from December 2017 to September 2020 were evaluated, and the efficacy assessment and survival analysis for high-risk patients and non-high-risk patients were evaluated.@*RESULTS@#After infusion of BCMA-CART cells in 21 MM patients, the number of effective cases was 17, of which the complete remission (sCR/CR) was 10, and the partial remission (VGPR/PR) was 7. The median OS time for all patients was 19.4 months, and the median PFS time was 7.9 months. The number of patients with extramedullary disease(EMD), high-risk genetics, and ISS stage Ⅲ were 5, 15 and 8, and the effective number was 3, 11 and 6, respectively. The treatment of 3 patients without high-risk factors was effective. The median OS and median PFS of patients with EMD were 14.2 and 2.5 months, respectively, which were shorter than those of patients without EMD (19.4 months and 8.9 months, respectively). The median OS and median PFS of patients with high-risk cytogenetic factors and ISS Ⅲ were not significantly different from those of non-high-risk patients. Cytokine release syndrane (CRS) occurred in 20 patients, of which 14 cases were Grade 1 CRS, while 6 were Grade 2, no CRS of Grade 3 or above occurred. IL-6 receptor inhibitors were used in 9 patients. All CRS were controlled effectively, and no patients had neurological toxicity.@*CONCLUSION@#BCMA-CART is a certain curative effect in the treatment of relapsed and refractory multiple myeloma, and the adverse reactions can be well controlled through close monitoring and timely treatment.

WU Xu's experience of sequential therapy for peripheral facial paralysis / 中国针灸

The paper introduces professor WU Xu 's experience of sequential therapy for peripheral facial paralysis. The sequential therapy refers to a staging treatment, but not rigidly adheres to it. With this therapy, the acupuncture- moxibustion regimen is modified flexibly in line with the specific symptoms of illness. At the acute phase of peripheral facial paralysis, warm acupuncture at Wangu (GB 12) is predominated and electroacupuncture is not recommended at the acupoints on the face. At the recovery phase, warm acupuncture at Zusanli (ST 36) is the main therapy and electroacupuncture is applied to the acupoints on the face appropriately. Besides, for the intractable case, the tapping technique with plum-blossom needle or skin needle should be combined and exerted in the local affected region. At the sequelae phase, in order to shorten the duration of illness, depending on the different types of facial paralysis, i.e. stiffness type, spasmodic type and flaccid type, the corresponding needling techniques are provided, i.e. bloodletting and moxibustion, strong stimulation with contralateral acupuncture and the technique for promoting the governor vessel and warming up yang.

Endophthalmitis Caused by Mixed Infections in Silicone Oil-filled Eye:One Case Report / 中国医学科学院学报

The incidence of endophthalmitis after vitrectomy is extremely low,especially lower in silicone oil-filled eyes.Silicone oil exerts a toxic effect on the cell membranes of microorganisms and leads to the lack of nutrients.It is thus believed to inhibit the growth of bacteria and fungi.Endophthalmitis induced by mixed bacteria in silicone oil-filled eye has been rarely reported.We reviewed the clinical manifestations,diagnosis,and treatment of a patient with endophthalmitis caused by mixed infection of

An Extrapulmonary Manifestation of Coronavirus Disease 2019:Olfactory Dysfunction / 中国医学科学院学报

In addition to acute respiratory symptoms,coronavirus disease 2019(COVID-19)could cause olfactory dysfunction,which becomes the only clinical manifestation of COVID-19 in some cases.We review the epidemiological characteristics,pathological mechanism,screening value,treatment and prognosis of olfactory dysfunction in patients with COVID-19,aiming to achieve an in-depth understanding of the early diagnosis,quarantine,scientific treatment and prognosis of COVID-19.

Bioinformatics Analysis and Verification of Acute B-Lymphocytic Leukemia in Children with Isolated Bone Marrow Relapse / 中国实验血液学杂志

OBJECTIVE@#To analyze the hub genes affecting the solely bone marrow relapse of childish acute B-cell lymphoblastic leukemia (B-ALL).@*METHODS@#The high-throughput RNA sequencing data were downloaded from TCGA database, the differentially expressed genes were screened by DESeq2 package of R, and the differentially expressed genes were grouped by GO function enrichment analysis and KEGG pathway enrichment analysis. Further, the data of STRING database and Cytoscape software were used to construct protein interaction network, screen hub genes and highly interaction protein sub network, perform GO and KEGG analysis of the hub genes and protein sub network respectively. JASPAR database was used to screen the upstream transcription factor of the hub gene promoter. Survival analysis based on the expression of hub genes was performed with clinical information attached to TCGA database. The bone marrow samples and clinical data of the patients were collected, the analysis results of hub genes were verified through clinical samples.@*RESULTS@#847 differentially expressed genes were collected, including 813 up-regulated genes, 34 down-regulated genes, 11 hub genes were screened out. The results of survival analysis showed that RPS5、RPS15、RPL23、RPL35、RPS8、RPS27A、RPS3、RPL9、RPS21、RPS7 and RPL38 showed significant effect on the survival of the children, and ZNF460 might be involved in their regulation. The high expressions of RPS3, RPS15, RPS8, RPS27A, and RPS21 had been verified in clinical samples of solely bone marrow relapsed patients.@*CONCLUSION@#RPS3, RPS15, RPS8, RPS27A, RPS21 can be used as biomarkers to indicate the malignant event of solely bone marrow relapse, which may be regulated by ZNF460.

Analysis on the response measures and outcomes of four epidemics of infectious diseases in Shanghai based on a time axis / 上海预防医学

Objective To compare the response measures and outcomes of SARS-CoV(2003), H1N1 influenza(2009), H7N9 influenza(2013)and COVID-19(2020)in Shanghai and provide scientific evidence for the emergency response of public health emergencies. Methods We compared the response measures and outcomes of the four epidemics in Shanghai in the aspects of government response, prevention and control system, scientific and technological support, social mobilization and prevention effects, using critical incident analysis based on the time axis of the epidemics from literature review. Results In response to the four epidemics of infectious diseases occurred in 2003-2020, Shanghai has generally made some significant effort and flexible measures in the first month of the epidemics, including"closure of live poultry markets"and"three closed-loops and four 100% coverage"and other specific prevention and control measures, which have enhanced the prevention and control system. However, we identified that the construction of prevention and control system for public health emergencies remained inefficient, compared to rapid economic development. The majority of the construction measures were principally post-epidemic. In addition, there were many challenges, such as passive response, temporary response teams and measures, and difficulties in the flexible bottom-up response for residents, families and industries without standard operating procedure and guidelines. Conclusion It may be an effective measure for the prevention and control of infectious diseases to build joint prevention and control measures with mass participation and regular drills, in the perspectives of strategic, tactical and operational levels of epidemic prevention and control.

Integrated 3D bioprinting-based geometry-control strategy for fabricating corneal substitutes / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

BACKGROUND@#The shortage of donor corneas is a severe global issue, and hence the development of corneal alternatives is imperative and urgent. Although attempts to produce artificial cornea substitutes by tissue engineering have made some positive progress, many problems remain that hamper their clinical application worldwide. For example, the curvature of tissue-engineered cornea substitutes cannot be designed to fit the bulbus oculi of patients.@*OBJECTIVE@#To overcome these limitations, in this paper, we present a novel integrated three-dimensional (3D) bioprinting-based cornea substitute fabrication strategy to realize design, customized fabrication, and evaluation of multi-layer hollow structures with complicated surfaces.@*METHODS@#The key rationale for this method is to combine digital light processing (DLP) and extrusion bioprinting into an integrated 3D cornea bioprinting system. A designable and personalized corneal substitute was designed based on mathematical modelling and a computer tomography scan of a natural cornea. The printed corneal substitute was evaluated based on biomechanical analysis, weight, structural integrity, and fit.@*RESULTS@#The results revealed that the fabrication of high water content and highly transparent curved films with geometric features designed according to the natural human cornea can be achieved using a rapid, simple, and low-cost manufacturing process with a high repetition rate and quality.@*CONCLUSIONS@#This study demonstrated the feasibility of customized design, analysis, and fabrication of a corneal substitute. The programmability of this method opens up the possibility of producing substitutes for other cornea-like shell structures with different scale and geometry features, such as the glomerulus, atrium, and oophoron.

Association between iron deficiency and prevalence of thyroid autoimmunity in pregnant and non-pregnant women of childbearing age: a cross-sectional study / 中华医学杂志(英文版)

BACKGROUND@#Thyroid autoimmunity (TAI) is prevalent among women of reproductive age and associated with adverse pregnancy outcomes. This study aimed to investigate the association between iron nutritional status and the prevalence of TAI in women during the first trimester of pregnancy and in non-pregnant women of childbearing age.@*METHODS@#Cross-sectional analysis of 7463 pregnant women during the first trimester of pregnancy and 2185 non-pregnant women of childbearing age nested within the sub-clinical hypothyroid in early pregnancy study, a prospective collection of pregnant and non-pregnant women's data, was conducted in Liaoning province of China between 2012 and 2015. Serum thyrotropin, free thyroxine, thyroid peroxidase antibodies (TPOAbs), thyroglobulin antibodies (TgAbs), serum ferritin, and urinary iodine were measured. Iron deficiency (ID) was defined as serum ferritin 150 μg/L. TPOAb-positive was defined as >34 U/mL and TgAb-positive was defined as >115 U/mL. Multilevel logistic regression was conducted to examine the association between TAI and different iron nutritional status after adjusting for potential confounders.@*RESULTS@#The prevalence of isolated TPOAb-positive was markedly higher in women with ID than those without ID, in both pregnant and non-pregnant women (6.28% vs. 3.23%, χ = 10.264, P = 0.002; 6.25% vs. 3.70%, χ = 3,791, P = 0.044; respectively). After adjusting for confounders and the cluster effect of hospitals, ID remained associated with TPOAb-positive in pregnant and non-pregnant women (odds ratio [OR]: 2.111, 95% confidence interval [CI]: 1.241-3.591, P = 0.006; and OR: 1.822, 95% CI: 1.011-3.282, P = 0.046, respectively).@*CONCLUSION@#ID was associated with a higher prevalence of isolated TPOAbs-positive, but not with isolated TgAb-positive, in both pregnant women during the first trimester of pregnancy and non-pregnant women of childbearing age, while IO was not associated with either isolated TPOAb-positive or isolated TgAb-positive.@*CLINICAL TRIAL REGISTRATION@#ChiCTR-TRC-13003805, http://www.chictr.org.cn/index.aspx.

Association between iron deficiency and prevalence of thyroid autoimmunity in pregnant and non-pregnant women of childbearing age: a cross-sectional study / 中华医学杂志(英文版)

Background@#Thyroid autoimmunity (TAI) is prevalent among women of reproductive age and associated with adverse pregnancy outcomes. This study aimed to investigate the association between iron nutritional status and the prevalence of TAI in women during the first trimester of pregnancy and in non-pregnant women of childbearing age.@*Methods@#Cross-sectional analysis of 7463 pregnant women during the first trimester of pregnancy and 2185 non-pregnant women of childbearing age nested within the sub-clinical hypothyroid in early pregnancy study, a prospective collection of pregnant and non-pregnant women’s data, was conducted in Liaoning province of China between 2012 and 2015. Serum thyrotropin, free thyroxine, thyroid peroxidase antibodies (TPOAbs), thyroglobulin antibodies (TgAbs), serum ferritin, and urinary iodine were measured. Iron deficiency (ID) was defined as serum ferritin <15 μg/L and iron overload (IO) was defined as ferritin >150 μg/L. TPOAb-positive was defined as >34 U/mL and TgAb-positive was defined as >115 U/mL. Multilevel logistic regression was conducted to examine the association between TAI and different iron nutritional status after adjusting for potential confounders.@*Results@#The prevalence of isolated TPOAb-positive was markedly higher in women with ID than those without ID, in both pregnant and non-pregnant women (6.28% vs. 3.23%, χ2 = 10.264, P = 0.002; 6.25% vs. 3.70%, χ2 = 3,791, P = 0.044; respectively). After adjusting for confounders and the cluster effect of hospitals, ID remained associated with TPOAb-positive in pregnant and non-pregnant women (odds ratio [OR]: 2.111, 95% confidence interval [CI]: 1.241–3.591, P = 0.006; and OR: 1.822, 95% CI: 1.011–3.282, P = 0.046, respectively).@*Conclusion@#ID was associated with a higher prevalence of isolated TPOAbs-positive, but not with isolated TgAb-positive, in both pregnant women during the first trimester of pregnancy and non-pregnant women of childbearing age, while IO was not associated with either isolated TPOAb-positive or isolated TgAb-positive.@*Clinical trial registration@#ChiCTR-TRC-13003805, http://www.chictr.org.cn/index.aspx.

Clinical efficacy and pharmacoeconomics analysis of three targeted drugs in the treatment of non-small cell lung cancer / 中国基层医药

Objective@#To compared the clinical efficacy and pharmacoeconomics of gefitinib, erlotinib, icotinib in the treatment of non-small cell lung cancer (NSCLC), thus to provide guidance on drug selection for patients from safety, efficiency and economical aspects after medical reform.@*Methods@#The patients with NSCLC who admitted to the First Affiliated Hospital of China Medical University from 2014 to 2017 and treated by gefitinib, erlotinib, icotinib were selected (30 patients in each group). All the patients were diagnosed with NSCLC by tissue or cytology study and eventually disease progressed.The clinical efficacy of the three drugs was evaluated by retrospective analysis.Specially, cost-effectiveness and cost-utility were evaluated in terms of disease control rates(DCR) and quality adjusted life years(QALY), respectively.@*Results@#The results demonstrated that the progression free survival (PFS) of gefitinib, erlotinib and icotinib were 0.934 years, 1.079 years and 1.063 years, respectively.There was no statistically significant difference in PFS among the three groups(F=0.001, P=0.9 990). The major drug adverse reactions were rash, diarrhea and hepatic injury.The DCR of gefitinib, erlotinib and icotinib were 66.7%, 70.0% and 63.3%, respectively, the difference was statistically significant(χ2=0.300, P=0.8 607). The total cost of treatment in the three groups were 85 118 CNY, 70 513 CNY and 7 2213 CNY, respectively.The cost-utility ratios of three drugs were 91 133, 65 389 and 67 973, respectively.The cost-effectiveness ratios of three drugs were 1 276, 1 007 and 1 141, respectively.The erlotinib group had lower cost and higher utility (effectiveness). Sensitivity analysis showed that the conclusion was relative stable when the price of three drugs changed at the same time.@*Conclusion@#All of the three drugs are applicable for targeted therapy of NSCLC and show similar curative effect and adverse reaction.However, erlotinib shows better economy than others.

Role of monocyte chemoattractant factor∕chemokine C-C receptor 2 pathway in amygdala in neuro-pathic pain in rats / 中华麻醉学杂志

Objective To evaluate the role of monocyte chemotactic factor-1 (MCP-1)∕chemokine C-C receptor 2 ( CCR2) in amygdala in neuropathic pain ( NP) in rats. Methods Clean-grade healthy male Sprague-Dawley rats, weighing 200-260 g, aged 2 months, in which NP model was established by ligating the left L5,6spinal nerve, were used in this study. The experiment was performed in two parts. Ex-periment Ⅰ Thirty-two rats were divided into 2 groups using a random number table method: control group (C group, n＝8) and NP group (n＝24). Rats were sacrificed at 7, 14 and 21 days after establis-hing NP model in group NP or at the corresponding time points before establishing NP model in group C, and the amygdala was removed for determination of the expression of MCP-1 and CCR2 mRNA and positive cell count using quantitative real-time polymerase chain reaction and immunohistochemistry. Experiment ⅡThirty-two rats were divided into 4 groups ( n＝8 each) using a random number table method: control group (C group), NP group, MCP-1 group and specific CCR2 antagonist RS102895 group (RS group). MCP-1 (50 pmol for each side) or RS102895 (100 pmol for each side) was injected into the bilateral a-mygdala at days 3, 6, 13 and 20 after establishing NP model in MCP-1 and RS groups, respectively. The thermal paw withdrawal latency (TWL) and mechanical paw withdrawal threshold (MWT) were measured at days 4, 7, 14 and 21 after establishing NP model (T1-4). Rats were sacrificed at T4and the L5segment of the spinal cord was removed for determination of interleukin-1beta (IL-1β), IL-6 and tumor necrosis fac-tor-alpha ( TNF-α) contents by enzyme-linked immunosorbent assay. Results Experiment Ⅰ Compared with group C, the expression of MCP-1 and CCR2 mRNA in amygdala was significantly up-regulated, and the number of MCP-1 and CCR2 positive cells was increased in group NP ( P＜0. 05). Experiment ⅡCompared with group C, the MWT was significantly decreased and TWL was shortened at T1-4, and the contents of IL-1β, IL-6 and TNF-α were increased in the other three groups ( P＜0. 05). Compared with group NP, the MWT was significantly decreased and TWL was shortened at T1, and the contents of IL-1β, IL-6 and TNF-α were increased in group MCP-1, and the MWT was significantly increased and TWL was prolonged at T1-4, and the contents of IL-1β, IL-6 and TNF-α were decreased in group RS ( P＜0. 05 or 0. 01). Conclusion Enhanced function of MCP-1∕CCR2 in amygdala may be involved in the pathophysio-logical process of NP in rats.

Rapid Identification of Oligosaccharide Isomers by Electron Capture Dissociation in Combination with Metal Ions / 分析化学

Oligosaccharide isomers were distinguished by electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry ( ECD-FT-ICR-MS ) in combination with utiliZing alkali, alkaline earth, and transition metals ( Na+, Ca2+, Ba2+, Mg2+, Mn2+ and Co2+) as charge carriers in electrospray.Maltoheptaose, mannohexaose and laminarihexaose were taken as examples to investigate influence of metal ions on the extent of oligosaccharide fragmentation.The same types of fragmentation ions ( 0,2 A and 2,4 A) were obtained for barium- and calcium-adducted maltoheptaose.Mg2+ and Mn2+ had the similar influence ( 0,2 A, 2,4 A and 2,5 A ).Three cross-ring cleavage ions ( 1,4 A, 2,4 A and 2,5 A ) were generated in the spectrum of cobalt-associated maltoheptaose.But in the case of doping Na+into maltoheptaose, only 0,2 A ion was detected.It was found that the signals in the spectra of mannohexaose and laminarihexaose were worse than that in the spectrum of maltoheptaose, probably resulting from different numbers of adducted metal ions.The isomers, mannohexaose and laminarihexaose could be distinguished by ECD-MS in conjunction with the addition of Ca2+, Mg2+ or Co2+.The addition of Ca2+ was the best choice for analysis of oligosaccharides.

Study on role of miR-208b-3p in dexmedetomidine alleviating rat myocardial ischemia-reperfusion injury / 重庆医学

Objective To investigate the role of miR-208b-3p in dexmedetomidine(DEX) alleviating rat myocardial ischemia reperfusion injury(MIRI).Methods Eighty adult male Wister rats were divided into the sham operation(Sham) group,I/R group,DEX group and miR-208b-3p+ DEX group.In the Sham group,a 6-0 silk suture was only placed without ligation,and other 3 groups were treated by I/R model.Before ischemia reperfusion,the DEX group received a loading dose of DEX(5 μg/kg) via the femoral vein followed by a continuous infusion of 5 μg · kg-1 · h-1 for 1 h.In the miR-208b-3p+ DEX group,rats received intramuscular injection of miR-208b-3p analogue at 24 h before ischemia reperfusion.The cardiac function indexes were monitored at 120 min after reperfusion,including heart rate(HR),left ventricular systolic blood pressure(LVSP),left ventricular end diastolic pressure(LVEDP) and left ventricular rate maximum(dp/dtmax).The serum levels of cTn-Ⅰ and CK-MB were detected and the apoptosis rate(AI) was measured by in situ apoptosis assay.The levels of oxidative stress related indicators were detected and Western blot was used to detect the level of myocardial apoptosis protein.Results Compared with the Sham group,the cardiac function in the I/R group was decreased,but the levels of CTn-Ⅰ,CK-MB,AI,MDA,Bax and Caspase-3 were increased(P＜0.05);compared with the I/R group,the above indexes in the DEX group were improved(P＜0.05);compared with the DEX group,the above indexes in the miR-208b-3p+DEX group were deteriorated(P＜0.05).Conclusion Overexpression of miR-208b-3p can eliminate the protective effect of DEX on MIRI.

Regulation of Intracellular Calcium by Endoplasmic Reticulum Proteins in Small Intestinal Interstitial Cells of Cajal

BACKGROUND/AIMS: We investigated the role of representative endoplasmic reticulum proteins, stromal interaction molecule 1 (STIM1), and store-operated calcium entry-associated regulatory factor (SARAF) in pacemaker activity in cultured interstitial cells of Cajal (ICCs) isolated from mouse small intestine. METHODS: The whole-cell patch clamp technique applied for intracellular calcium ions ([Ca²+]i) analysis with STIM1 or SARAF overexpressed cultured ICCs from mouse small intestine. RESULTS: In the current-clamping mode, cultured ICCs displayed spontaneous pacemaker potentials. External carbachol exposure produced tonic membrane depolarization in the current-clamp mode, which recovered within a few seconds into normal pacemaker potentials. In STIM1-overexpressing cultured ICCs pacemaker potential frequency was increased, and in SARAF-overexpressing ICCs pacemaker potential frequency was strongly inhibited. The application of gadolinium (a non-selective cation channel inhibitor) or a Ca2+-free solution to understand Orai channel involvement abolished the generation of pacemaker potentials. When recording intracellular Ca²+ concentration with Fluo 3-AM, STIM1-overexpressing ICCs showed an increased number of spontaneous intracellular Ca²+ oscillations. However, SARAF-overexpressing ICCs showed fewer spontaneous intracellular Ca2+ oscillations. CONCLUSION: Endoplasmic reticulum proteins modulated the frequency of pacemaker activity in ICCs, and levels of STIM1 and SARAF may determine slow wave patterns in the gastrointestinal tract.

Effect of Compound Jiegu Tablets and Zhuanggu Tablets on the differentiation of bone marrow mesenchymal stem cells in vitro / 中国组织工程研究

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are a kind of pluripotent stem cells in the bone marrow. Drug-containing serum treatment may be followed by a variation in the expression of genes involved in multiple signaling pathways. OBJECTIVE: To study the effect of compound Jiegu Tablets and Zhuanggu Tablets on the differentiation of BMSCs in vitro. METHODS: Sixty Sprague-Dawley rats were randomized into Jiegu group, Zhuanggu group, combined group and blank control group. Different drug-containing sera were prepared and used to culture BMSCs isolated from the rat bone marrow. RT-PCR and western blot assay were used to detect the expression of BMP-2, Runx2, VEGF and Akt at mRNA and protein levels, respectively, at 24 hours after culture.RESULTS AND CONCLUSION: Compared with the blank control group, the expression levels of Akt and VEGF mRNA and protein were significantly increased in the Jiegu group and combined group (P < 0.05 or P < 0.01), but showed no difference in the Zhuanggu group (P > 0.05). Compared with the blank control group, the expression levels of BMP-2 and Runx2 mRNA and protein were significantly increased in the Jiegu group (P < 0.05) and very significantly increased in the Zhuanggu group and combined group (P < 0.01). These findings indicate that compound Jiegu Tablets and Zhuanggu Tablets containing sera lead to synergistic effects on the BMSCs differentiation into vascular endothelial cells and osteoblasts in vitro.

Relationship between neuropeptide S in amygdala and 5-hydroxytryptamine and GABA in spinal dorsal horns of rats with neuropathic pain / 中华麻醉学杂志

Objective To evaluate the relationship between neuropeptide S (NPS) in the amygdala and 5-hydroxytryptamine (5-HT) and GABA in spinal dorsal horns of rats with neuropathic pain.Methods Eighty pathogen-free healthy male Sprague-Dawley rats,weighing 200-260 g,aged 2 months,were divided into 4 groups (n =20 each) using a random number table:sham operation group (group Sham),neuropathic pain group (group NP),low dose NPS group (group L-NPS) and high dose NPS group (group H-NPS).The neuropathic pain model was established by left L5,6 spinal nerve ligation (SNL) in anesthetized rats.NPS was injected into the bilateral amygdala at 3,6,9,12,15 and 18 days after SNL in LNPS group (10 pmol per side) and H-NPS group (100 pmol per side).The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 days before SNL and 1,4,7,11,14,17 and 21 days after SNL.Five rats were selected at 7,14 and 21 days after SNL and sacrificed,and the lumbar segment (L5) of the spinal cord was removed for detection of the expression of 5-HT and GABA in spinal dorsal horns by immunofluorescence histochemistry.Results Compared with group Sham,the MWT was significantly decreased,the TWL was shortened,and the expression of 5-HT and GABA in spinal dorsal horns was down-regulated in NP,L-NPS and H-NPS groups (P＜0.05).Compared with group NP,the MWT was significantly increased,the TWL was prolonged,and the expression of 5-HT and GABA in spinal dorsal horns was up-regulated in L-NPS and H-NPS groups (P＜0.05).Compared with group L-NPS,the MWT was significantly increased,the TWL was prolonged,and the expression of 5-HT and GABA in spinal dorsal horns was up-regulated in group H-NPS (P＜0.05).Conclusion The spinal mechanism of endogenous analgesia induced by NPS in the amygdala may be related to up-regulation of the expression of 5-HT and GABA in spinal dorsal horns of rats with neuropathic pain.

Content comparsion of six alkaloids in different Zingiber-processed Aconiti lateralis Radix Praeparata / 中成药

AIM To compare the contents of benzoyl mesaconitine,benzoyl hypaconitine,benzoyl aconitine,aconitine,hypaconitine and mesaconitine in Zingiberis Rhizoma Recens boiled juice-processed,Zingiberis Rhizoma boiled juice-processed,Zingiberis Rhizoma Recens juicing-processed,Zingiberis Rhizoma Recens slice malaxation steaming,and Zingiberis Rhizoma slice malaxation steaming Aconiti lateralis Radix Praeparata.METHODS Acid-base titration method was adopted in the content determination of total alkaloids.HPLC was applied to determining the contents of six alkaloids,the analysis was performed on a 35 ℃ thermostatic Waters Symmetry(C)C1scolumn (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.04 mol/L ammonium acetate flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 235 nm.RESULTS The contents of total alkaloids and diester-type alkaloids (mesaconitine,aconitine and hypaconitine)in various processed products,especially for Zingiberis Rhizoma Recens slice,Zingiberis Rhizoma slice malaxation steaming products,were obviously lower than those in the raw product.The contents of benzoyl hypaconitine and benzoyl aconitine in Zingiberis Rhizoma slice malaxation steaming product were the lowest,while that of benzoyl mesaconitine in the raw product was the lowest.CONCLUSION Compared with other Zingiber-processing methods,both Zingiberis Rhizoma Recens slice and Zingiberis Rhizoma slice malaxation steaming can reduce the toxicity of Aconiti lateralis Radix Praeparata more effectively.

Targeted Genes Sequencing Identified a Novel 15 bp Deletion on GJA8 in a Chinese Family with Autosomal Dominant Congenital Cataracts / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Congenital cataract (CC) is the leading cause of visual impairment or blindness in children worldwide. Because of highly genetic and clinical heterogeneity, a molecular diagnosis of the lens disease remains a challenge.</p><p><b>METHODS</b>In this study, we tested a three-generation Chinese family with autosomal dominant CCs by targeted sequencing of 45 CC genes on next generation sequencing and evaluated the pathogenicity of the detected mutation by protein structure, pedigree validation, and molecular dynamics (MD) simulation.</p><p><b>RESULTS</b>A novel 15 bp deletion on GJA8 (c.426_440delGCTGGAGGGGACCCT or p. 143_147delLEGTL) was detected in the family. The deletion, concerned with an in-frame deletion of 5 amino acid residues in a highly evolutionarily conserved region within the cytoplasmic loop domain of the gap junction channel protein connexin 50 (Cx50), was in full cosegregation with the cataract phenotypes in the family but not found in 1100 control exomes. MD simulation revealed that the introduction of the deletion destabilized the Cx50 gap junction channel, indicating the deletion as a dominant-negative mutation.</p><p><b>CONCLUSIONS</b>The above results support the pathogenic role of the 15 bp deletion on GJA8 in the Chinese family and demonstrate targeted genes sequencing as a resolution to molecular diagnosis of CCs.</p>