Objective To study the protective effects of Tianma xingnao capsule on brain blood circulation in mice and rats.Methods The arteria carotis externa was ligated,and arteria carotis interna was kept.The left carotid artery was exposed and placed over an ultrasonic flow probe.The brain flow volume was recorded with an Ultrasonic Volume.The pursiness time of mice was observed to evaluate the effect of Tianma xingnao capsule on cerebral ischemia and anoxia.The permeability of normal blood-cerebral barrier was investigated in mice.Results Tianma xingnao capsule significantly increased brain blood flow and prolonged mouse pursiness time.Tianma xingnao capsule at a dose of 4 g/kg significantly elevated the content of Evans in mouse brain.Conclusions Tianma xingnao capsule can increase the brain blood flow in rats and improve brain blood circulation in mice.It is obviously advantageous to protect against cerebral ischemia and hypoxia.

Objective:To investigate the correlation between lumbar bone mineral density (BMD), musculoskeletal perfusion andmuscle mass.Methods:From May 2019 to August 2020, totally 91 patients who applied for CT perfusion (CTP) examination of abdomen (the scan range included the vertebral body of L1－L3) in Shanghai Tenth People′s Hospital of Tongji University were retrospectively analyzed. The mean BMD of L1－L3 vertebral body was measured by quantitative CT (QCT) at the same time of CT plain scan. According to BMD, the subjects were divided into normal BMD group ( n=33), osteopenia group ( n=41) and osteoporosis (OP) group ( n=17). The L3 level perivertebral muscle mass index and fat fraction were calculated based on QCT examination. The lumbar vertebral and perivertebral muscle perfusion parameters were measured based on CTP images. The parameters of QCT and CTP among three groups were analyzed by Kruskal-Wallis H test or one-way ANOVA. The correlation analysis was conducted between these parameters using Pearson or Spearman analysis. Results:The differences of the perivertebral muscle mass index and fat fraction among three groups were statistically significant ( P<0.05). The differences of the lumbar vertebral perfusion parameters including blood flow (BF), blood volume (BV) and flow extraction product (FE) among three groups were statistically significant ( P<0.05), and BF, BV and FE were positively correlated with BMD ( r=0.444, 0.312 and 0.266 respectively, all P<0.05; adjusted for age and gender r=0.437, 0.340 and 0.337 respectively, all P<0.05). There was no statistically significant difference in perivertebral muscle perfusion parameters among three groups ( P>0.05). Perivertebral muscle mass index was negatively correlated with fat fraction ( r=-0.599, P<0.001; adjusted for age and gender r=-0.404, P<0.001), and there was no correlation between perivertebral muscle mass index and muscle perfusion parameters, as well as perivertebral muscle fat fraction and muscle perfusion parameters. Conclusions:With the changes of BMD, bone mass and perivertebral muscle mass at L3 level are synchronous. Decreased vertebral bone mass is accompanied with reduced perivertebral muscle mass, increased muscle fat and decreased bone perfusion. The changes of vertebral perfusion and perivertebral muscle perfusion at L3 level are asynchronous, which implies that reduced perfusion in OP patients may be confined to the bone.

Objective To analyze the clinical features of infantile cytomegalovirus(CMV) hepatitis with cholestasis and investigate intrahepatic cholestasis due to hepatocytic impairment caused by CMV infection.Methods Forty-seven children with CMV cholestatic he-patitis were divided into 2 groups according to the level of total bilirubin(TB):22 cases with serum TB lower than 136.8 ?mol/L(groupⅠ),and 25 cases with serum TB higher than 136.8 ?mol/L(groupⅡ).All children were treated with both gangciclovir and routine met-hods,and serum biochemistry were checked before and after treatment.SPSS 13.0 software was used to analyze the data.Results Forty-seven cases of infantile CMV cholestatic hepatitis had different degrees of jaundice,hepatosplenomegaly and abnormal liver functions.The differences of serum ALT and AST between the 2 groups had statistical significance,the levels of serum gamma glutamy transferase(GGT) and alkaline phosphatase(ALP) were lightly higher in groupⅡcompared with those in groupⅠ,but there were no statistical significance.TB,direct bilirubin(DB),ALT and AST were decreased in the 2 groups after treatment,GGT and ALP hadn′t decreased significantly after treatment.Conclusions CMV infection can injure hepatocytes and epithelials on each grade of bile duct,thus CMV hepatitis causes intrahepatic cholestasis.Cholestasis due to hepatocytic impairment deserves emphasis and intervention should be done as early as possible.Gangciclovir therapy for CMV infection manifest effective and safe in short-term.

Objective To report a peliosis hepatic in child and review literature and discuss.Methods Case history was inquired.Physical,labtoratory,imagement and histopathology of liver biopsy(HE staining) were examed.Results A 4-year old girl appeared dermatitis with erythema and herpes at local skin where was bit by insect before onset.The girl appeared fever,cough,then abdominal pain,hepatomegaly,pleural effusion and ascites.Lab examination revealed slight elevation of aspartate transaminase,?-glutamyltranspeptidase and alkaline phosphatase.The liver B-mode ultrasonography and CT scan revealed hepatomegaly with density heterogeneity of the parenchyma.The liver biopsy revealed many small capsule filled with blood cells.Conclusions Clinical characteristics of the disease are fever,upper abdomen pain,janundice,ascites and hepatomegaly.The diagnosis shall be combined with the pathologic biopsy of liver.

OBJECTIVETo evaluate the feasibility and efficacy of a new MRI imaging method--diffusion weighted imaging (DWI) with short TI version recovery-echo planar imaging (STIR-EPI) sequence in differentiating benign cervical lymph nodes from malignant ones. METHODS Twenty nasopharyngeal carcinoma patients and fourteen volunteers received both conventional MRI and DWI with STIR-EPI. Ability of detecting lymph nodes between conventional MRI and STIR-EPI-DWI was compared, and the difference of apparent diffusion coefficient (ADC) value between metastatic lymph node and normal lymph node was analyzed.

RESULTSDWI was more sensitive in detecting lymph node than conventional MRI. ADC value of metastatic lymph node (0. 766 +/- 0. 119) x 10 (-3) mm(2)/s was significantly lower than that of normal lymph node (0. 975 +/- 0. 179) x 10 - mm2/s (P < 0. 01).

CONCLUSIONAs a new MRI imaging technique in detecting cervical lymph nodes, diffusion weighted imaging ( DWI) with short TI version recovery-echo planar imaging ( STIR-EPI) sequence is more reliable and sensitive than conventional MRI imaging, providing an alternative way to differentiate benign lymph nodes from malignant ones.

Adult ; Aged ; Diagnosis, Differential ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; methods ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; Neck ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo study the modulatory effect of insulin on apoptosis of skeletal myoblast (L6 cells) by serum of scalded rat and its mechanism.

METHODSL6 cells cultured with DMEM medium containing 10% FBS were divided into control (C, added with 20% normal rat serum), serum from rat with scald injury (S, added with 20% serum from scalded rat), insulin (I, added with 20% normal rat serum and 100 nmol/L insulin), and serum of scalded rat + insulin (SI, added with 20% serum of scalded rat + 100 nmol/L insulin) groups according to the random number table. After being cultured for 48 hours, apoptosis was observed with Hoechst 33258 staining and its number counted, annexin V -FITC/PI double-labeling method was used to assess apoptosis rate, the protein levels of phosphorylated (p-) Akt, p-PI3K, Bax, Bcl-2, and active caspase-3 were determined by Western blotting. Data were processed with grouped or paired t test.

RESULTS(1) The amount of apoptosis with typical morphological change in S group [(59.6 +/- 3.9) per visual field] was more than that in C, I, and SI groups [(4.9 +/- 2.6), (5.5 +/- 2.1), (19.7 +/- 2.3) per visual field, with t value respectively 28.53, 29.86, 21.53, P values all below 0.01]. (2) Apoptotic rate in S group was (18.5 +/- 1.8)%, which was markedly higher than that in C, I, and SI groups [(1.1 +/- 0.6)%, (1.5 +/- 0.3)%, (7.8 +/- 0.6)%, with t value respectively 22.41, 22.83, 13.92, P values all below 0.01]. (3) Compared with those in C group, the protein levels of Bax and active caspase-3 in S group were up-regulated (1.12 +/- 0.63 vs. 0.16 +/- 0.03, 2.15 +/- 0.51 vs. 0.21 +/- 0.03, with t value respectively 3.80, 10.69, P values all below 0.01), the protein level of p-Akt was lowered (0.20 +/- 0.03 vs. 0.42 +/- 0.07, t = -8.46, P < 0.01), and the protein levels of p-PI3K and Bcl-2 showed no statistical difference (0.19 +/- 0.03 vs. 0.26 +/- 0.09, 0.17 +/- 0.03 vs. 0.28 +/- 0.07, with t value respectively -2.73, - 1.14, P values all above 0.05). The protein levels of Bax (0.40 +/- 0.14) and active caspase-3 (0.83 +/- 0.18) in SI group were lowered (t = -3.23, P < 0.05; t = 6.66, P < 0.01) and the protein levels of p-Akt, Bcl-2, and p-PI3K in SI group were elevated (0.39 +/- 0.10, 0.78 +/- 0.03, 0.47 +/- 0.12, with t value respectively 4.07, 18.71, 5.05, P < 0.05 or P < 0.01) as compared with those in S group.

CONCLUSIONSSerum from scalded rat can induce apoptosis in skeletal myoblast, and the effect can be inhibited by insulin through PI3K/Akt signal pathway.

Animals ; Apoptosis ; drug effects ; Burns ; blood ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line ; Insulin ; pharmacology ; Male ; Myoblasts, Skeletal ; cytology ; drug effects ; pathology ; Rats ; Rats, Wistar ; Serum ; immunology ; Signal Transduction ; bcl-2-Associated X Protein ; metabolism

A strain, MMR003, used for D-p-HPG production in industry was classified as Burkholderia pickettii by morphological observation and biochemical characterization. The gene encoding the D-hydantoinase enzyme was cloned, sequenced and expressed in Escherichia coli. The nucleotide sequence of the 5.0 kb insert of subclone pXZ-total was determined. One open reading frame of 1374 bp was found and predicted to encode a polypeptide consisting of 458 amino acids in size of 50 kD. The amino acid sequence alignment of D-hydantoinase from Burkholderia pickettii shows the 85% homologous with the corresponding enzyme from Agrobacterium radiobacter NRRL B11291. The D-hydantoinase gene (dha) harboured in the plasmid pXZPH2 in E. coli BL21(DE3) was highly expressed by IPTG induction. The D-hydantoinase activity for D, L-p-hydroxyphenylhydantion is 0.66 u/mL broth, which is 2-fold increase compared to the parent strain Burkholderia pickettii.
Amidohydrolases ; genetics ; metabolism ; Amino Acid Sequence ; Animals ; Burkholderia ; classification ; enzymology ; genetics ; Cloning, Molecular ; DNA Probes ; Escherichia coli ; Gene Expression ; Molecular Sequence Data ; Recombination, Genetic ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

OBJECTIVETo investigate the apoptosis-inducing effect of trichloroethylene (TCE) on cultured normal human epidermis keratinocytes (NHEK) in vitro.

METHODSNR(50) values (the concentration of neutral red absorbed is reduced to 50%) of TCE on NHEK were assayed by neutral red uptake (NRU), and the administered dose of TCE was determined. Lipid peroxidation (LPO) and oxidative stress were assessed by measurement of malondialdehyde (MDA) contens and superoxide dismutase (SOD) activity. Transmission electron microscope (TEM) were used to observe morphologic changes, flow cytometer (FCM) was used to measure DNA contents and calculate cell apoptosis rate and proliferation index (PI).

RESULTSNR(50) values of TCE on NHEK was found to be 4.53 mmol/L (95% CI: 3.92-5.13 mmol/L). The increase in MDA content and inhibition of SOD activity in a concentration-dependent manner were shown after NHEK was treated with a series of dose of TCE 4 h later, and typical morphologic changes of apoptosis were also observed by TEM examination. FCM analysis revealed a sub-G(1) peak in the apoptotic cells. The apoptotic rate in TCE 0.125, 0.500, 2.000 mmol/L exposed groups (31.83%, 38.63%, 44.35%, respectively) were significantly higher than that in blank control (18.42%), while PI in TCE 0.125, 0.500, 2.000 mmol/L group (3.26%, 2.48%, 2.07%, respectively) were significantly lower than that in blank control (4.99%).

CONCLUSIONTCE may induce apoptosis of cultured NHEK in vitro, and inhibit cell proliferation through lipid peroxidation and oxidative stress.

Apoptosis ; drug effects ; Cells, Cultured ; Epidermis ; cytology ; drug effects ; Humans ; Keratinocytes ; drug effects ; Lipid Peroxidation ; Oxidative Stress ; Trichloroethylene ; toxicity

OBJECTIVETo explore mechanism of dermal toxicity of trichloroethylene(TCE).

METHODSNormal human keratinocytes (KC) were isolated from foreskins of healthy donors undergoing circumcision by two-step trypsin digestion and cultured in serum-free medium. Cells were treated with medium, 1% acetone (volume fraction) 0.125, 0.500 or 2.000 mmol/L TCE for different time (4, 8, 12 or 24) hours. After treatment, MTT assay and ATPase activity detected, inhibition ratio of mitochondrial enzyme was calculated according to optical density (A) value of MTT assay. Mitochondrial membrane potential (MMP) was detected by flow cytometry FCM after being stained with Rhodamine123 (Rh123). Morphological changes were also observed through transmission electron microscope (TEM).

RESULTSCellular viability and ATPase activity declined with dose of TCE, while inhibition ratio of mitochondrial enzyme increased with dose of TCE. FCM results showed that after treatment with 2.000 mmol/L TCE, fluorescence density of Rh123 decreased quickly from 18.73 +/- 0.45(0 h) to 8.20 +/- 0.66(8 h) (P < 0.01). After 8 h, fluorescence density maintained at the level equal to that of 8 h (fluorescence density of Rh123 were 8.20 +/- 0.36 and 8.20 +/- 0.40 for 12 and 24 h respectively, compared with that for 8 h group, P > 0.05). The results also showed that MMP diminished with dose of TCE. Under TEM, mitochondria in TCE-treated group appeared extensive swelling and vacuolar degeneration with less matrix and obscure or vanished mitochondria cristae but in control group, mitochondrial structure was integrated, with uniform matrix and visible mitochondria cristae.

CONCLUSIONSTCE could inhibit mitochondrial metabolic enzyme, reduce ATP production, diminish MMP, and destroy ultrastructure of mitochondria in KC, all these contributing to the cytotoxicity of TCE.

Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Keratinocytes ; drug effects ; metabolism ; ultrastructure ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Microscopy, Electron, Transmission ; Mitochondria ; drug effects ; metabolism ; ultrastructure ; Trichloroethylene ; toxicity

OBJECTIVESTo compare expressions of tyrosine-phosphorylated proteins in different hepatocellular carcinoma cell lines with different metastasis potential and to screen key molecules associated with HCC metastasis and recurrence.

METHODSUsing two-dimensional electrophoresis, Western blotting and MALDI-TOF-MS/MS, we analyzed tyrosine-phosphorylated protein profiles of Hep3B, MHCC97L and MHCC97H, HCC cell lines with different metastasis potentials.

RESULTS10 spots were detected in Hep3B, 19 in MHCC97L and 17 in MHCC97H. Seventeen significantly different phosphotyrosine proteins in gel were identified by MALDI-TOF-MS/MS, including Annexin I.

CONCLUSIONThe changed expression of tyrosine-phosphorylated proteins is associated with HCC metastasis and recurrence.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Neoplasm Proteins ; analysis ; Phosphotyrosine ; analysis