Objective: To study the suspension culture of tuber and its alkaloid content based on the stimulation of salicylic acid. Method: The tubers of Pinelliae Rhizoma in suspension tube were treated with different concentrations of exogenous salicylic acid to analyze the growth status. The content of alkaloids in tuber was detected by HPLC. Test conditions:chromatographic column for Agilent Eclipse plus C₁₈ column (4.6 mm×250 mm,5 μm),mobile phase of acetonitrile water(4:96),the column temperature was maintained at 35℃,detection wavelength for inosine 250 nm,guanosine 260 nm,volume flow rate 1.0 mL·min^-1. Result: The results showed that the exogenous salicylic acid had a certain effect on the growth of suspension tuber of Pinelliae Rhizoma. When the salicylic acid concentration was 150 μmol·L^-1,the culture lasted for 25 days and the fresh weight reached the maximum value of 7.483 8 g. It also accumulates a certain amount of alkaloids. The linear range of guanosine was 0.03-0.45 μg (R²=0.999 6). After 10-days cultivatation in the salicylic acid concentration of 50 μmol·L^-1,guanosine content of Pinelliae Rhizoma tubers reached a maximum of 1.353 3 mg·g^-1. The linear range of inosine 0.003-0.045 μg (R²=0.999 5). When the salicylic acid concentration was 200 μmol·L^-1,cultured for 30 days,the content of inosine in Pinelliae Rhizoma tubers reached the maximum value of 0.149 8 mg·g^-1. Conclusion: The results of this experiment provide a reference for the study of tissue culture and rapid propagation of Pinelliae Rhizoma tubers and regulation of alkaloids,which are of great significance for the development of Pinelliae Rhizoma industry.

Objective To explore the clinical effect of peroneal artery perforator flap for the treatment of serious heel spoke injuries in children.Methods From July,2014 to September,2017,13 children with severe heel and ankle injuries were treated by peroneal artery perforator flap.There were 9 males and 4 females,with an average age of 6.5 years (ranged from 3 to 11 years).The size of flaps ranged from 10.0 cm×3.0 cm to 14.5 cm×5.0 cm.The donor sites were sutured directly or covered with skin grafting.Regular follow-up was performed to observe the survival status of the flap and the functional recovery of the ankle joint.Results Twelve flaps survived completely.One flap had partial marginal necrosis in the distal portion,which was healed after dressing.Partial inadequate venous return happened in 1 case,which also recovered by the removal of part of the suture.All cases were followed-up for 3 months to 20 months.The appearance,texture,and color of the flaps were similar to the surrounding skin.No ulcer occurred.All case acquired normal gait.Conclusion The peroneal artery perforator flap is a good option for reconstruction of serious heel spoke injuries in children.The flap has consistent blood supply while leaving minimal morbidity at donor site.

Objective To explore the effect of hypergravity on morphology and osteogenesis function of preosteoblast MC3T3-E1 ceils.Methods The cultured MC3T3-E1 cells under hypergravity by different loading forces were divided into five groups,including control group,5 g group,10 g group,15 g group and 20 g group.The experimental groups were loaded for 30 min each time in 3 successive days,and the control group with no g-value was synchronously exposed to the same surrounding.The morphology of cytoskeletal protein was observed by phalIoidin staining,The alkaline phosphatase (ALP) content was examined by ALP activity assay kit,the gene expression of ALP,collagen Ⅰ (Col Ⅰ),osteocalcin (OC),runt-related transcription factors (Runx2) was measured by real-time quantitative PCR,and the protein expression of Col Ⅰ and OC was tested by Western blotting.Results Under the condition of hypergravity,cell body of osteoblast became thinner,but its surface area increased significantly;with the structure of skeletal arrangement becoming loose,actin microfilament structure reduced so that the orderly arrangement of actin-like dispersion lowered.The gene expressions of related indicators of osteogenic differentiation including ALP,Col][,OC,Runx2 were significantly up-regulated,which was the same as Col Ⅰ protein and OC protein after hypergravity loading.A very minute quantity of small red-orange nodules was found in the control group,while the cells in experimental groups after hypergravity loading obviously formed various sizes of red-orange nodules.Conclusions Under hypergravity,changes in osteoblast morphology can be triggered by rearrangements of skeletal structure.Furthermore,osteoblast maturation and differentiation can be stimulated effectively by up-regulating differentiation-related gene and protein expressions.

Objective To explore the effect of hypergravity on morphology and osteogenesis function of preosteoblast MC3T3-E1 cells. Methods The cultured MC3T3-E1 cells under hypergravity by different loading forces were divided into five groups, including control group, 5 g group, 10 g group, 15 g group and 20 g group. The experimental groups were loaded for 30 min each time in the three successive days, and the control group was synchronously exposed to the same surrounding except for difference in g-value. The morphology of cytoskeletal protein was observed by phalloidin staining, The alkaline phosphatase (ALP) content was examined by ALP activity assay kit, the gene expression of ALP, collagen Ⅰ(ColⅠ), osteocalcin (OC), runt-related transcription factors (Runx2) was measured by real-time quantitative PCR, and the protein expression of ColⅠ and OC was tested by Western blot. Results Under the condition of hypergravity, cell body of osteoblast became thinner, but its surface area increased significantly; with the structure of skeletal arrangement becoming loose, actin microfilament structure reduced so that arrangement of actin-like dispersion orderly lowered. The gene expressions of related indicators of osteogenic differentiation including ALP, ColⅠ, OC, Runx2 loaded by hypergravity were significantly up-regulated, which was the same as ColⅠ protein and OC protein after hypergravity loading. There was only a very minute quantity of small red-orange nodules in the control group, while the cells after hypergravity loading in experimental groups obviously formed various sizes of red-orange nodules. Conclusions Under hypergravity, changes in osteoblast morphology can be triggered by rearrangements of skeletal structure. Furthermore, osteoblast maturation and differentiation can be stimulated effectively by up-regulating differentiation-related gene and protein expressions.

Objective To discuss the clinical features diagnosis and treatment of cerebral schistosomiasis. Methods A to?tal of 166 patients with cerebral schistosomiasis were treated and their clinical data were collected and analyzed retrospectively. Results In 166 cases of cerebral schistosomiasis the confirmative diagnoses of 156 cases were diagnosed according to the clin?ical manifestation etiology immunology and auxiliary examination CT MRI .In among 74 cases were confirmed by pathologi?cal examination 10 cases were diagnosed through to the diagnostic treatment. Totally 102 patients received the oral medication of praziquantel and they all improved and discharged 14?16 days later 64 patients received the craniotomy and praziquantel medication after the operation and 48 patients significantly improved others did not improve or aggravated. There was no opera?tive mortality. Conclusions Neuroimaging and laboratory tests are valuable in the diagnosis of cerebral schistosomiasis. The praziquantel treatment is selected firstly when the diagnosis was established. However in the case of serious intracranial hyper?tension intractable epilepsy and praziquantel treatment fails the surgical treatment is required.

OBJECTIVETo investigate the regulation of different hypoxia on cell survival and autophagy.

METHODSPC12 cells were treated with different hypoxia. The cell survival was measured by MTT assay, expressions of LC3 and p62 were marked for autophagy detected by Western Blot, and the level of reactive oxygen species (ROS) was analyzed by flow cytometry.

RESULTSThe cell viability was different under different hypoxia: moderate hypoxia promoted cell viability, and severe hypoxia caused a decrease in cell viability; autophagy marker molecules, p62 and LC3-II expressions were different: moderate hypoxia increased p62 and LC3-II expressions, in contrast, severe hypoxia led to the decrease of p62 and LC3-II expressions; compared to normoxia, moderate hypoxia did not change the levels of ROS, while severe hypoxia increased the levels; 3-MA, the inhibitor of autophagy, elevated the levels of ROS in the three oxygen concentrations, additionally, the increased amplitudes in the moderate and severe hypoxia groups were higher than that in the normoxia group.

CONCLUSIONModerate hypoxia promotes cell survival, severe hypoxia causes the cell death, and the autophagy activity may mediate the effects of different hypoxia.

Animals ; Autophagy ; physiology ; Cell Death ; Cell Hypoxia ; Cell Survival ; PC12 Cells ; Rats ; Reactive Oxygen Species ; metabolism

BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.

METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.

RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.

CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.

Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

OBJECTIVETo observe the effects of acute hypoxia on the cell adhesion molecule close homologue of L1 (CHL1) expression in different brain areas and main organs (heart, lung, kidney) of mice, and provide a basis for the role of CHL1 in hypoxia injury.

METHODSMice were randomly divided into two groups (n=10): normoxia group and hypoxia group. Hypoxia group were treated by acute hypoxia (8% O2, 8 h). Protein expression changes in different tissues were evaluated by Western blot.

RESULTSIn central nervous system, CHL1 protein expressions were down-regulated in cerebral cortex, hypothalamus and brain stem by acute hypoxia and up-regulated in cerebellum. In heart and lung, CHL1 protein expression were down-regulated by acute hypoxia.

CONCLUSIONCHL1 protein expressions were changed in different tissues after acute hypoxia, which suggested CHL1 might play an important role in hypoxia damage regulation.

Animals ; Brain ; metabolism ; Cell Adhesion Molecules ; genetics ; metabolism ; Hypoxia ; metabolism ; Lung ; metabolism ; Male ; Mice ; Myocardium ; metabolism ; Tissue Distribution

OBJECTIVERecent study demonstrated that hypoxia could regulate the proliferation and differentiation of neural stem cells in vitro. In the present study, effects of low glucose and/or hypoxia on the proliferation and metabolism of neural stem cells were investigated in vitro.

METHODSThe neural stem cells were isolated from the rat embryonic mesencephalon (E13.5), and exposed to different oxygen concentrations (low oxygen: 3% O2 or normoxia: 20% O2) and different glucose concentrations (high glucose concentration: 4.5 g/L and low glucose concentration: 1.4 g/L) for 3 days. The proliferation of neural stem cells were examined by CCK-8 assay. Furthermore, the content of glucose, lactate, and pyruvic acid in the medium were measured after cultured in different condition for 1, 3, 5 days.

RESULTSLow oxygen and low glucose could increase the proliferation of neural stem cells respectively; in addition, the number of neurospheres under both low oxygen and glucose was the most among the four groups. The content of glucose and pyruvic acid in the medium from low oxygen or low glucose condition decreased, while the lactate concentration increased compared with the control group.

CONCLUSIONThe results indicate the neural stem cells prefer grow under the low glucose and low oxygen condition, and that is mainly under going glycolysis to maintain its self-renew ability. This study may provide us a useful clue for application of neural stem cells transplantation.

Animals ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Female ; Glucose ; metabolism ; Glycolysis ; Neural Stem Cells ; cytology ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the prevalence of J wave in apparently healthy subjects in Wuhan.

METHODSThe study subjects comprised of 1817 apparently healthy subjects (1131 males, mean age 46.38 +/- 15.81 years; 686 females, mean age was 42.77 +/- 14.15 years). ECG and routine medical examinations were performed. J wave was defined as a wave followed QRS complexes with amplitude of at least 0.05 mV and 0.03 s.

RESULTSThe overall incidence of J wave was 7.26%. The incidence of J wave in males was significantly higher than in females (10.53% vs. 1.87%, P < 0.01). The incidence of J wave in leads of inferior wall (II, III, avF), right wall (V(1 - 3)) and left wall (V(4 - 6)) was 4.57%, 0.50%, and 2.20%, respectively. J wave located in leads of inferior wall was more than in left and right walls (both P < 0.05). The incidence of J wave positively correlated with age (y = 0.1387x + 1.6318, r = 0.78, P < 0.01).

CONCLUSIONSJ wave is more likely seen in males and aged people and is more likely located in leads of inferior wall, than in leads of left and right walls.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Death, Sudden, Cardiac ; epidemiology ; prevention & control ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Physical Examination ; Prevalence ; Young Adult