BACKGROUND: BALB/c and C57BL/6 mice are two inbred strains, but after voluntary movement, there is no report on how to scientifical y reasonably select behavioral experiment methods and indicators and to evaluate the learning and memory abilities of mice.
OBJECTIVE: To analyze and compare the behavioral indicators between BALB/c and C57BL/6 mice fol owing voluntary wheel running, to explore the effect of exercise on learning and memory, and to provide a reference for selecting reasonable behavioral indicators.
METHODS: 2.5-month-old BALB/c and C57BL/6 mice were randomly divided into control and voluntary wheel running groups. Independent running wheel movement of mice was recorded with VitalView system. 4 weeks later, newborn neurons were labeled via DCX immunofluorescence. Spatial learning, memory and exploration abilities were compared through new arm test, new object recognition test and Morris water maze test.
RESULTS AND CONCLUSION: (1) The mean spontaneous activity of BALB/c mice daily was 2.56 fold of that of C57BL/6 mice during wheel running (P < 0.001). (2) Hippocampal DCX-positive cel s in exercised BALB/c and C57BL/6 mice were more than those in control group. (3) Meantime, exhibited by higher frequencies to explore new arm or object, and longer time and distance of moving around them, the learning and exploring capability was improved after exercising (P < 0.001), especial y in BALB/c mice. (4) Wheel running C57BL/6 mice exhibited progressed spatial learning and memory abilities compared with control mice in Morris water maze test, characterized by decreased latency to target, elevated target crossings and longer time or distance in quadrant zone (P < 0.05). However, there was no significant difference between wheel running and control BALB/c mice. Taken these data together, voluntary wheel running contributed to hippocampal neurogenesis of BALB/c and C57BL/6 mice, accompanied by the change of learning and memory capability, which could be detected properly via both new arm test and new object recognition test, but for Morris water maze test, C57BL/6 mice might be superior to BALB/c mice.

Objective:To investigate the heterogeneity of epitopes recognized by anti-GBM autoantibodies in sera from a large cohort of Chinese patients with anti-GBM disease and its clinical significance.Methods: The present study included 108 patients with anti-GBM disease who were diagnosed in our hospital, between Jan 1991 and May 2009, with complete clinical and renal pathological data. Sera or plasma exchange of the patients were used to incubate with cryostat section of normal human renal tissue for indirect immunofluorescence (IIF) assay. The cryostat sections of normal renal tissue were pre-treated by 6 mol/L urea to unmask cryptic epitopes, and untreated cryostat sections were used to detect natural exposed epitopes. The sera were diluted from 1:2 to 1:512 to determine titers of anti-GBM autoantibodies Patients with anti-GBM autoantibodies against cryptic or exposed epitopes were further stratified;their clinical and pathological associations were analyzed. Results: Sera from all the 108 patients could recognize cryptic epitopes on normal renal tissue ( urea treated section). IIF showed IgG linear staining along GBM. However, sera from 56/108 patients (group A) could also recognize exposed epitopes on normal renal tissue (untreated section) ; sera from the rest 52/108 patients (group B) could not recognize exposed epitopes. In urea treated condition, the average titer of anti-GBM autoantibodies from sera of patients in group A was significantly higher than that in group B (P＜0.01) , ANCA-positive patients in group A were significant less than that in group B (P＜0.01) . There was no significant difference between the two groups in regard to other clinical data (including serum creatinine) and renal histopathologic data. Conclusion: Anti-GBM autoantibodies from some patients with anti-GBM disease could recognize natural exposed epitopes, however, their anti-GBM titer for cryptic epitopes was higher than that of those recognizing cryptic epitopes only and the prevalence of serum ANCA was significantly less.

Objective To assess the treatment outcomes of anterior segment optical coherence tomography(OCT) assisted excimer laser phototherapeutic keratectomy (PTK) for fungal corneal ulcer.Methods Twenty-four patients (24 eyes) suffered from fungal corneal keratitis were collected by a retrospecive chart review with at least 6 months of follow-up data available.All corneal ulcer depth did not exceed 1/3 corneal thickness and were resistant to local and systemic medication therapy for at least 1 week.Patients were divided into 2 groups.14 patients with 14 eyes were taken PTK and 10 patients with 10 eyes were treated with lamellar keratectomy.Follow-up time was at least 6 months.Visual acuities,corneal thickness,confocal microscope,healing of the epithelial defect and inflammation and complications were recorded.Results The postoperative visual acuity in two groups were significantly improved,visual acuity of PTK treatment group improved significantly better than the control group (P ＜ 0.05).Healing time of PTK treatment group (19.43 ± 5.68) days was shorter than that of the control group (34.40 ± 5.38) days,the difference was statistically significant (P ＜0.05).There was no significant difference in the residual central corneal thickness between the PTK treatment group and the control group (P ＞ 0.05).Residual mycelium after operation in the treatment group was significantly decreased than that of control group.Healing time of epithelial in PTK treatment group (4.0 ± 1.1) days was shorter than that of the control group (6.20 ± 1.39) days,the difference was statistically significant (P ＜ 0.05).There was no corneal dilation in two groups.No recurrence in PTK treatment group was occurred.3 cases in control group had recurrence after 6 months,drug control was effective in 1 case,and other 2 cases undertook corneal transplantation.Conclusion Anterior segment OCT assisted PTK for corneal ulcer can effectively clear the lesion,accelerate epithelial healing,provide a new choice for the treatment of early superficial fungal corneal ulcer.

Objective To evaluate the value of MR in the diagnosis of primary splenic lymphoma (PSL).Methods The MR imaging features of 3 PSL cases proved by pathology were retrospectively reviewed. Results Three cases were all pathologically diagnosed as B-cell non-Hodgkin's lymphoma(NHL). Two cases were multiple node shape and one was massive shape. Unenhanced MR imaging revealed heterogeneous splenic enlargement, with large nodules showing iso-hyperinternse on T1WI and hypointense on T2WI. Linear hyperintense on T1WI and T2WI was seen on the spleen peripherous. The spleen vascular was infiltrated. CE-MRI showed heterogeneous enhancement of spleen with iso-hypointense. The focus of postperitoneal showed medial enhancement. Immunohistochemistry-showed 2 cases of diffuse B cell. The CD20 and CD19 α of tumor cell showed diffuse( + ) ,CD3 ,CD5 individual( + ) ,CD43 ( + ) ,CD45RO( + ), CD10 ( +/- ), Mum ( + ), MAC387 individual ( + ), 1 case of B lymphocell type, CD79α ( + ) ,CD23( + ) ,CD38( + ) ,λ( + ). Conclusion The MR imaging features of PSL were characteristic and helpful in the diagnosis of PSL, but the correct diagnosis was still dependent on the pathology and immunohistochemical staining.

Objective:To understand the clinical characteristics and prognosis of Langerhans cell histiocytosis (LCH) with skin-limited lesion.Methods:A retrospective analysis was performed on clinical characteristics and prognosis of 16 skin-limited LCH patients, out of 578 LCH patients who were hospitalized in Beijing Children′s Hospital during December 2013 to June 2018.Results:A total of 16 skin-limited LCH cases, accounted for 2.7% of all 578 cases, were included.Among which, sex ratio (male vs.female) was 1.28∶1.00.Median ages of skin eruption occurrence and of diagnosis of the disease were 3.5 months (3 days to 2 years and 5 months) and 6 months (2 months 14 days to 2 years and 8 months) in this group.Among the 16 cases, seborrheic dermatitis-like lesions(11 cases, 68.7%) was the most common, and the trunk was most frequently involved[75.0% (12 cases)]. Serine/threonine protein kinase gene V600E [ BRAF (p.V600E)] mutation was detected in pathological specimens from 10 skin-limi-ted cases, with 9 cases being positive.Plasma samples from 5 positive cases were further detected for BRAF (p.V600E) mutation, and 4 positive results were gained.Of all 16 patients, 11 cases (68.7%) were treated.Remission were achieved in 3-6 months from treatment start in patients treated whether according to the Histiocyte Society′s LCH-2009 protocol for 25 weeks(6 cases, 37.5%), or with topical mometasonefuroate for 3 months (3 cases, 18.8%). Two patients(12.5%) with solitary cutaneous lesions underwent excision biopsy (one face and one prepuce) and were considered to be in remission immediately after surgery.None of these patients suffered from the recurrence of the disease.The remaining 5 patients (31.3%) with skin-limited LCH were just evaluated regularly, and achieved remission in 3-6 months of commencing observation.Among these untreated patients, 1 with consistently positive BRAF (p.V600E) mutation in plasma had bone involvement in the 24 th month of assessment, and was then treated based on the Histiocyte Society′s LCH-2009 Protocol.No clinical or imageological evidence supporting disease progression was found on this patient.Median follow-up period was 32.8 months (2.9-63.9 months). Except one patient, none of the rest cases had active disease till follow-up ended.Two-year event free survival(EFS) of this research was (92.3± 7.4)%.There was no significant difference between EFS of treated group and that of observation group( χ2=1.250, P=0.264). Conclusions:Skin-limited LCH often occurs in infants and newborns, with strong heterogeneity in clinical manifestations, laboratory indicators, and pathogenesis.Seborrheic dermatitis-like lesions were the most common cutaneous type.The prognosis of the patients is excellent despite progressing into multisystem involvement can be seen in a few patients.

In recent years, the prevention and cure of infectious bursal disease (IBD) have become more and more difficult due to the emergence of very virulent strains of infectious bursal disease virus (vvIBDV) and the variant strains of IBDV. In this research, the hybridoma cell lines which secretes anti-idiotypic antibodies against anti-IBDV IgG were established. According to the Jerne's theory of immune network, the use of the anti-idiotypic antibodies as a vaccine will be a new method for the prevention of IBD. In this study, the SPF chickens were inoculated with the IBDV- SD strain, and the bursal was obtained from the died chickens. The bursal was then homogenized and frozen-thawed 3 cycles, and the virus samples were prepared by cane sugar density gradient centrifugation and dialysis. Typical IBDV particles were observed under an electron microscope, and the concentration of the virus protein measured by ultraviolet absorbance spectrophotometry was 10.8 mg/mL. SPF chickens were immunized with the virus and the highly immunized sera were prepared and purified by Sulfuric acid ammonia salt out and Sephadex G-25 chromatography. Then, Balb/C mice of six or eight weeks old were immunized interapertoneally(I. P.) with purified antibodies to IBDV at regular intervals. SP2/0 myeloma cells were fused with the spleencytes from the immunized mice at a ratio of 10:1, in 50% polyethylene glycol (1540) and were then cultured in HAT until all the SP2/0 cells died. The hybridoma cells were selected by ELISA and the highly positive holes were cloned 3 times with the method of limited dilution. Two strains (2B6 strain,5F4 strain) of hybridoma cells were obtained, which were shown by ELISA to steadily secrete anti-IBDV idiotypic antibodies. The chromosome number of the two hybridoma cells were about 88 - 106, 95 in average, and the antibodies secreted belonged to the types of IgG1 and Kappa. Balb/c mice of 3 months old were inoculated I.P. with about 10(7) hybridoma cells per capita, and the ascites were collected 12 days later and the titre of anti-IBDV idiotypic antibodies measured by ELISA was 1 :25600 (for 2B6) and 1:12800 (for 5F4) . The ascites containing the anti-IBDV idiotypic antibodies were emulsified with complete or incomplete Freund's adjuvants, and the anti-IBDV idiotypic antibody vaccine was obtained. SPF and common Jingbai chickens were immunized with the vaccine obtained. The immunized chickens with the vaccine were inoculated with IBDV-SD strain at a dose of 2000 ELD50 after twoimmunizations. All the 10 SPF chickens in the non-immunized group were sick, and 8 of them died; and 5 out of the 50 SPF chickens immunized group got sick and 2 died. All the 10 common Jingbai chickens in the control group were sick, and 6 died; 7 of the 30 immunized common Jingbai chickens got sick and only 1 died. Chi2 analysis showed that the difference between the immunized and the non-immunized groups in both the SPF and the common Jingbai chickens were significant (P < 0.01). Our result indicated that the anti-IBDV idiotypic antibody vaccine well protected chickens and had a great potential in both research and clinical application.
Animals ; Antibodies, Anti-Idiotypic ; biosynthesis ; immunology ; Birnaviridae Infections ; immunology ; Cell Line, Tumor ; Chickens ; Enzyme-Linked Immunosorbent Assay ; Hybridomas ; immunology ; metabolism ; Infectious bursal disease virus ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; Viral Vaccines ; biosynthesis ; immunology

OBJECTIVETo investigate the effects of dopamine (DA) and metabolite in striatum of Parkinson's disease (PD) rats treated by bone mesenchymal stem cells (MSCs) modified by plasmid pIRESneo-EGFP-BDNF.

METHODpIRESneo-EGFP-BDNF was transfected to MSCs with electroporation. The rat models of PD were set up by 6-OHDA and then divided into four groups randomly, which were Sham group, PD group, BDNF group. The rotating behavior of rat models induced by apomorphine (APO) intraperitoneally which transplanting bone MSCs or MSCs modified by plasmid pIRESneo-EGFP-BDNF through cerebral lateral ventricle after 2, 4 and 8 weeks. The levels of DA, homovanillic acid (HVA), dihydroxy-phenylacetic acid (DOPAC) were measured by high performance liquid chromatography (HPLC) in striatum of each group.

RESULTSThe rotation numbers (r/min) of MSCs group or BDNF group in the 2nd, 4th and 8th week after transplanting were significantly decreased compared with that of PD group (P < 0.05). Those of BDNF group were specially significant compared with those of MSCs group (P < 0.05). The levels of DA, HVA, DOPAC and the ratios of DA/HVA, DA/DOPAC in stratum after PD rats intervened by transplanting cells through cerebral lateral ventricle after eight weeks were increased significantly in BDNF group or MSCs group while compared with PD group, especially in BDNF group.

CONCLUSIONThe behavior of rat with PD was improved significantly by increasing the levels of DA and decreasing metabolic rate of DA in striatum while transplanting BDNF modified bone MSCs through cerebral lateral ventricle.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; Corpus Striatum ; metabolism ; Disease Models, Animal ; Dopamine ; metabolism ; Genetic Engineering ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Parkinson Disease ; metabolism ; Plasmids ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the clinical effect of plantar medial perforator artery based reverse island medial dorsal pedal neurocutaneous vascular flaps.

METHODS12 cases with soft tissue defects of forefeet were treated by plantar medial perforator artery based reverse island medial dorsal pedal neurocutaneous vascular flaps. The flap size ranged from 3.0 cm x 3.5 cm to 5.5 cm x 8.5 cm.

RESULTAll flaps survived completely. The patients were followed up for 6 - 24 months. The texture and flexibility of the flaps were normal with no ulcer. The sensation improved with the two-point discrimination of 7 - 10 mm. The cosmetic and functional results were satisfactory. The wounds at donor site healed primarily.

CONCLUSIONSThe flaps have expanded size for large defects with good flexibility, thickness and texture. It is easily performed with less morbidity to main artery.

Adult ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Middle Aged ; Skin Transplantation ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; innervation ; Tibial Arteries ; surgery ; Young Adult

The metabonomics method was used to study the intervention effect of Psoraleae Fructus and Myristicae Semen in "Ershen pill" on the changes in serum endogenous metabolites in spleen-kidney Yang deficiency diarrhea rats before and after processing, screen out differentiated metabolites related to spleen-kidney Yang deficiency diarrhea and explore the metabolic patterns related to spleen-kidney Yang deficiency diarrhea and the processing synergy mechanism of Psoraleae Fructus and Myristicae Semen in "Ershen pill". Efforts were made to detect SOD and MDA of each group, test rat serum metabolic fingerprints in different stages by using GC-MS, analyze by PCA and PLS-DA methods and screen out potential biomarks through VIP and t test. The results revealed that "Ershen pill" could enhance the level of SOD and decrease the level of MDA and identified 10 differentiated metabolites related to spleen-kidney Yang deficiency diarrhea. Compared with the model group, all of metabolites recovered to varying levels after being intervened with "Ershen pill", with the best effect shown in the "Ershen pill" IV group (salt-processed Psoraleae Fructus + bran-roasted Myristicae Semen). It is speculated that that Psoraleae Fructus and Semen Myristicae in "Ershen pill" show a synergistic effect by inhibiting peroxide, improving aglucolipid, amino acids and energy metabolism, with multiple target sites.
Animals ; Chemistry, Pharmaceutical ; Diarrhea ; drug therapy ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Energy Metabolism ; drug effects ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Metabolomics ; Myristicaceae ; chemistry ; Psoralea ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spleen ; drug effects ; metabolism ; Yang Deficiency ; drug therapy ; metabolism

Objective To evaluate the effect of cardiovascular disease (CVD) on the healthcare costs associated with chronic kidney disease (CKD).Methods A total of 96 patients with stage 3-4 CKD treated at CKD clinic of Beijing Friendship Hospital,Capital Medical University were enrolled in the study.Their mean age was (61.6±9.5) years including 65 male and 31 female patients.A retrospective analysis of clinical material and health claims between January 2010 and October 2010 was conducted.Firstly,patients were grouped according to median CKD-associated healthcare cost and clinical characteristics were compared between two groups.Secondly,patients were stratified into three categories based on CVD prevalence (with acute cardiovascular events,with CVD but no acute events,and without CVD),and CKD-associated healthcare costs were assessed among the groups.Finally,the potential factors influencing CKD-associated healthcare costs were evaluated by optimal scaling regression analysis.Results During January to October in 2010,median CKD-associated healthcare costs was 13960.5 yuan (interquartile range 10226.5,19667.2 yuan).In the group with higher healthcare costs,more females,greater urine albumin-creatinine ratio,more emergency observations and/or hospitalizations caused by acute cardiovascular events,higher diabetes mellitus prevalence and calcium-phosphorus products,and lower eGFR and hemoglobin levels were found (P＜0.05,respectively).In contrast,the total prevalence of CVD was not significantly different between the groups (P=0.386).When grouping by CVD prevalence,significant differences of CKD-associated healthcare costs were observed only between patients with acute cardiovascular events and the other two groups (P＜0.01,respectively).The median healthcare cost of the former was approximately twice as higher as that of the other two groups,and the maximal cost was also found in the acute-cardiovascular-event group.For the optimal scaling regression analysis,both emergency observations and/or hospitalizations caused by acute cardiovascular events and diabetes mellitus entered the equation,and standardized coefficients were -0.538 and -0.217 respectively (P＜0.01 and P＜0.05).Conclusions Emergency observations and/or hospitalizations caused by acute cardiovascular events are important factors inducing high CKD-associated healthcare costs in patients with stage 3-4 CKD.Therefore,the prevention of acute cardiovascular events may be favorable to reduce CKD-associated healthcare costs.Larger and longer-time perspective studies are required to confirm it.In addition,diabetes mellitus also influences CKD-associated healthcare costs.