Objective To study the spatial arrangement of mouse connecting tubules（CNT）and their transition to collecting duct system.Methods The renal tissues of three C57/BL/6J mice were fixed by perfusion and embedded in Epon 812.Totally 2 000 consecutive sections with the thickness of 2.5 μm were obtained from the renal surface to the papilla.The serial images under microscope were digitalized and aligned.Thirty eight CNT were traced with a series of computer programs.Results The CNT from long loop nephron ascended towards renal capsule and merged with 5 to 6 CNT from short loop nephrons to form the so-called arcade,while the latter or some of the CNT from the superficial cortex directly drained into the collecting duct in the superficial cortex.The lengths of CNT and the arcade ranged from 150 μm to 500 μm and 600 μm to 900 μm respectively.Conclusion The arcade or CNT drains into collecting duct only at superficial cortical level,which suggests that the reabsorption of the glomerular ultra-filtration in collecting ducts keeps unaffected when collecting duct runs through middle to deeper cortex.

Objective The use of in vivo cryotechnique (IVCT) in combination with electrocardiograph (ECG) to study cardiac microcirculation under different hemodynamic conditions in living mouse.Methods Living mouse heart monitored by electrocardiograph was suffered from IVCT and freezing substitution under normal blood flow,myocardial ischemia or cardiac arrest conditions.Hematoxylin eosin (HE)staining,Schiff's staining and immunofluorescence staining for serum albumin,immunoglobulin were utilized on continuous paraffin sections,respectively.Confocal microscopy and statistical analyses were used.Results Comparing with normal hemodynamics,microvascular red cell volume reduction,morphology changed,myocardial cell glycogen loss,serum albumin ectopic distribution to myocardial cytoplasm,T tubular network failure and spacing width were happened in myocardial iscbemia condition; different shapes of red blood cells,myocardial cells glycogen deficiency,T tubular network failure and interval narrowing were found under cardiac arrest conditions.Conclusions Cardiac microcirculation,pathological changes of myocyte and its surrounding microenviroument in living mouse heart can be immediately captured in situ by the application of IVCT and ECG.

Objective To investigate the expression of endocytic receptor megalin and cubilinin in the developing mouse kidneys, and the correlation between the expression and the development of the renal tubules. Methods Expression of megalin and cubilin in developing mouse kidneys was examined at different embryonic days (E) using immunohistochemistry. Meanwhile, the ultrastructure of developing proximal tubules related to endocytosis was observed at transimission electron microscope level. Results At E9.5, megalin and cubilin were co-expressed in apical plasma membrane of the mesonephric ducts and mesonephric tubules. From E11 to E18, in the metanephros, the expression of both receptors were seen at the free surface and apical plasma of uretic bud, but weakly in all renal tubules of S shaped body at early differentiatial stage. With the mature of proximal tubule development, they were both confined to the brush border and the apical plasma of the proximal tubules in juxtamedullary cortex.Conclusion The endocytic receptors, megalin and cubilin are expressed in apical part of nearly all renal tubule epithelia in early development, and confined to free surface of mature proximal tubules, suggesting that with the mature of proximal tubules, the two receptors are generally involved in collaborating to facilitate, the reabsorption of ultrafiltration.

Objective: To investigate the clinical features, imaging features, pathological features and gene diagnosis of cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). Methods: Clinical manifestations, signs and imaging characteristics of a female patient hospitalized in the First Affiliated Hospital of Zhengzhou University for more than 10 days due to headache were analyzed, and skin biopsy and HTRA1 and Notch3 gene detection were performed. The pedigree of the proband was investigated in detail, and HTRA1 gene test and related imaging examination were conducted in parallel. Due to the deceased parents of the patient, relevant genetic testing could not be conducted. A control group of 100 healthy people were analyzed. Results: The clinical manifestations of proband were headache after insomnia, hearing loss in the right ear, easy to wake up and sweat at night. Brain MRI showed diffuse patchy long T₁ and long T₂ signals in bilateral fronto-parietal temporal occipital insula, internal and external capsule areas, bilateral basal ganglia areas, and bilateral thalamus. Fluid attenuated inversion recovery sequence showed high signals. Magnetic sensitive weighted imaging showed scattered patchy low signals in bilateral cerebral and cerebellar hemispheres, bilateral basal ganglia area, left thalamus and brain stem. The proband had consanguineous parents. A homozygous mutation C to T transition at position 589 (c.589C>T) was found in exon 3 of HTRA1 gene with the proband and both siblings. The heterozygous c.589C>T mutation appeared in another sister of the proband. Under the light microscope of skin biopsy, pigmentation in the basal layer of the skin could be seen, collagen fiber hyperplasia in the dermis was accompanied by a small amount of inflammatory cell infiltration, and no definite amyloidosis was found. No mutations were found in Notch3 gene. Because the patient′s parents were deceased, genetic testing was not possible. One hundred healthy controls had no such mutation. Conclusions The CARASIL family with HTRA1 gene c.589C>T homozygous mutation was reported, and the pathogenicity of the mutation was confirmed. HTRA1 genetic testing is recommended for diagnosis and differential diagnosis of CARASIL with family history or clinical suspicion.

Objective To investigate the clinical features, imaging features, pathological features and gene diagnosis of cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). Methods Clinical manifestations, signs and imaging characteristics of a female patient hospitalized in the First Affiliated Hospital of Zhengzhou University for more than 10 days due to headache were analyzed, and skin biopsy and HTRA1 and Notch3 gene detection were performed. The pedigree of the proband was investigated in detail, and HTRA1 gene test and related imaging examination were conducted in parallel. Due to the deceased parents of the patient, relevant genetic testing could not be conducted. A control group of 100 healthy people were analyzed. Results The clinical manifestations of proband were headache after insomnia, hearing loss in the right ear, easy to wake up and sweat at night. Brain MRI showed diffuse patchy long T1 and long T2 signals in bilateral fronto?parietal temporal occipital insula, internal and external capsule areas, bilateral basal ganglia areas, and bilateral thalamus. Fluid attenuated inversion recovery sequence showed high signals. Magnetic sensitive weighted imaging showed scattered patchy low signals in bilateral cerebral and cerebellar hemispheres, bilateral basal ganglia area, left thalamus and brain stem. The proband had consanguineous parents. A homozygous mutation C to T transition at position 589 (c.589C>T) was found in exon 3 of HTRA1 gene with the proband and both siblings. The heterozygous c.589C>T mutation appeared in another sister of the proband. Under the light microscope of skin biopsy, pigmentation in the basal layer of the skin could be seen, collagen fiber hyperplasia in the dermis was accompanied by a small amount of inflammatory cell infiltration, and no definite amyloidosis was found. No mutations were found in Notch3 gene. Because the patient′s parents were deceased, genetic testing was not possible. One hundred healthy controls had no such mutation. Conclusions The CARASIL family with HTRA1 gene c.589C>T homozygous mutation was reported, and the pathogenicity of the mutation was confirmed. HTRA1 genetic testing is recommended for diagnosis and differential diagnosis of CARASIL with family history or clinical suspicion.

Objective : To explore the expression of chemokine CXCL1 in proliferative infantile hemangioma (IH) , and to study the effect of exogenous CXCL1 on hemangioma stem cells (HemSCs) . Methods : Immunohistochemistry was used to explore the expression of CXCL1 in proliferative IH specimens.Primary HemSCs were isolated from IH tissues by CD133 magnetic beads.5 groups of CXCL1 with different concentrations(0,10,20,50 and 100 ng/ml) were co-cultured with HemSCs, and the effects of exogenous CXCL1 on HemSCs were studied by cell viability and migration experiments. Results : CXCL1 was expressed in the interstitial tissues of proliferative IH.The overall expression of CXCL1 in proliferative IH was low, but the expression of CXCL1 in the proliferative IH interstitial tissues was higher than that of the adjacent interstitial tissues.The CXCL1 positive area rate was(0.773±0.101)% in the tumor compared with(0.268±0.081)% in the adjacent tumor, and the difference was statistically significant(t=7.843,P<0.001).Exogenous CXCL1 promoted the proliferation of HemSCs, and there were statistical differences after adding different concentrations of CXCL1 to HemSCs for 24,48,and 72 h(F=14.610,P<0.001;F=14.430,P<0.001;F=5.388,P<0.01).But the exogenous CXCL1 did not affect the migration ability of HemSCs. Conclusion The expression of CXCL1 in proliferative IH interstitial tissues is higher than that in adjacent interstitial tissues, and exogenous CXCL1 promotes the proliferation of HemSCs.