[Mechanism of action of Jieduhuayu granules for remission of oxidative stress in hepatocytes].

Objective: To study and explore the effect and mechanism of action of Jieduhuayu granules on oxidative injury of human liver L02 cells. Methods: Human liver L02 oxidative injury model was established with 0.1 mmol/ L H(2)O(2) intervention for 1 h, and treated with different concentrations of Jieduhuayu (JDHY) solution. Hepatocytes were divided into five groups: normal, H(2)O(2), H(2)O(2) + JDHY (0.5 mg/ml), H(2)O(2) + JDHY (1 mg/ml), and H(2)O(2) + JDHY (1.5 mg/ml). MTT assay was used to detect hepatocytes activity. Transmission electron microscope was used to observe mitochondrial morphology in hepatocytes. Biochemical test was used to detect the levels of superoxide dismutase, lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, malondialdehyde, and reduced glutathione and albumin in hepatocytes. Western blot was used to detect the expression levels of rabbit anti-phosphatidylinositol 3-kinase (PI3K), AKT and mTOR in hepatocytes. One-way analysis of variance was used for comparison between multiple groups, and the LSD method was used for pairwise comparison. Results: Compared with the normal group, the cell proliferation activity (P < 0.05), mitochondrial vacuolization, superoxide dismutase activity, reduced albumin and glutathione content, and PI3K, AKT, and mTOR protein expression levels in the H(2)O(2) group were all significantly reduced (P < 0.05), while the content of malondialdehyde and the activities of alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase were significantly increased (P < 0.05). Compared with H(2)O(2) group, the cell proliferation activity (P < 0.05), alterations in morphological remission of mitochondria, superoxide dismutase activity, reduced albumin and glutathione content, and PI3K, AKT and mTOR protein expression levels in the H(2)O(2) + JDHY (1 mg/ml) and H(2)O(2) + JDHY (1.5 mg/ml) group (P < 0.05) were all significantly increased (P < 0.05), while malondialdehyde content and alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities were significantly decreased (P < 0.05). Conclusion: Jieduhuayu granule can effectively improve oxidative stress and mitochondrial injury in hepatocytes, and its effect may be related to the promoting expression of PI3K/AKT/mTOR signaling pathways.

[Texture analysis based on contrast-enhanced MRI can predict treatment response to neoadjuvant chemotherapy of breast cancer].

Objective: To investigate whether texture analysis based on contrast-enhanced MRI can predict pathological complete response of locally advanced breast cancer undergoing neoadjuvant chemotherapy(NAC). Methods: Forty-seven patients with breast cancer undergone neoadjuvant chemotherapy from January 2015 to February 2016 were divided into pathological complete response (pCR) group or non-pathological complete response (non-pCR) group based on surgical pathology. Their parameters of texture analysis based on MRI before neoadjuvant chemotherapy and after 2 cycles of treatment were analyzed. Parameters(Energy, Entropy, Inertia, Correlation, Inverse Difference Moment)before and after 2 cycles of NAC between pCR and non-pCR groups were compared using Student t or Wilcoxon rank sum test. The diagnostic performance of different parameters was judged by the receiver-operating characteristic (ROC) curve analysis. Results: The post-NAC value was significantly different from that of pre-NAC (all P<0.05). Pre-treatment parameters (Energy, Entropy, Inertia, Correlation, Inverse Difference Moment) were 78.58×10(-5)(55.64×10(-5), 135.23×10(-5)), 10.06 ± 1.02, 7 993.91±2 428.10, (4.76±0.99) ×10(-5) and (18.10±4.13) ×10(-3) in pCR group, and 76.84×10(-5) (48.68×10(-5), 154.15×10(-5)), 10.28±1.26, 7 184.77 (4 938.03, 9 974.04), (5.21±2.01) ×10(-5) and (17.68±5.87) ×10(-3) in non-pCR group. No significant difference was found between both groups. (P>0.05 for all). At the end of the second cycle of NAC, parameters(Energy, Entropy, Inertia, Correlation, Inverse Difference Moment) were (542.11±361.04) ×10(-5,) 7.95±1.28, 16 765.08±97 06.56, (0.43±0.07) ×10(-5,) and (12.18±9.82) ×10(-3) in pCR group, and 133.00×10(-5) (79.80×10(-5,) 239.00×10(-5)), 9.29±1.46, 7 916.64(6 418.89, 10 934.40), (0.38±0.08) ×10(-5) and (14.80±5.06) ×10(-3) in non-pCR group. At the end of the second cycle of NAC, there was significant difference in the parameters (Energy, Entropy, Inertia, Correlation) and Δparameters (ΔEnergy, ΔEntropy, ΔInertia, ΔInverse Difference Moment) between both groups (P<0.05 for all). The area under curve (AUC) of post-treatment ΔEntropy was 0.81, which was the largest one among parameters. Sensitivity of ΔEntropy for predicting pCR was 75.0% and specificity was 85.7%, respectively. Conclusion: Texture analysis based on dynamic contrast-enhanced MRI can predict early treatment response in primary breast cancer.

Structure and expression of fibulin-2, a novel extracellular matrix protein with multiple EGF-like repeats and consensus motifs for calcium binding.

A new protein, fibulin-2, was predicted from sequence analysis of cDNA clones obtained from a mouse fibroblast library. This protein consists of a 1195-residue polypeptide preceded by a 26-residue signal peptide. The COOH-terminal region of 787 amino acids contained three anaphylatoxin-related segments (domain I), 11 EGF-like repeats (domain II), 10 of which had a consensus motif for calcium-binding, and a 115-residue globular domain III. Except for two additional EGF-like repeats, this COOH-terminal region showed 43% sequence identity with the previously described fibulin-1 (BM-90). The NH2-terminal 408 residues, unique to fibulin-2, showed no sequence homology to other known proteins and presumably form two additional domains that differ in their cysteine content. Recombinant fibulin-2 was produced and secreted by human cell clones as a disulfide-bonded trimer. Rotary shadowing visualized the protein as three 40-45 nm long rods which are connected at one end in a globe-like structure. No significant immunological cross-reaction could be detected between fibulin-1 and fibulin-2. Production of the fibulin-2 was demonstrated by Northern blots and radioimmunoassay in fibroblasts but not in several tumor cell lines. Together with the observation that the serum level of fibulin-2 is 1,000-fold lower than that of fibulin-1, the data indicate that these two isoforms are not always coordinately expressed. This is also suggested by Northern blots of tissue mRNAs and by immunofluorescence localizations using mouse tissues. The latter studies also demonstrated an extracellular localization for fibulin-2 in basement membranes and other connective tissue compartments.

The fluorescence recovery of polydiacetylene/fluorophore vesicles by interaction with cetyltrimethylammonium bromide.

Polydiacetylene (PDA) has been widely used as a biosensor candidate because it exhibits specific blue-red transition upon varied external stimulations, such as heat, pH, or mechanical pressure. We prepared mixed vesicles using the 10,12-pentacosadiynoic acid (PCDA) and fluorescence probe BO558, and investigated the interaction of surfactant with PCDA/BO558 vesicles. Experiments exhibited that the fluorescence of BO558 was significantly quenched owing to the ene-yne conjugated backbones in the PCDA vesicles. However, the fluorescence was gradually recovered in the presence of cetyltrimethylammonium bromide (CTAB) to the system. Meanwhile, the turbidity of the mixed solution decreased greatly with the increase of CTAB concentration. We investigated the mechanism of the fluorescence quenching and recovery in the system of PCDA/BO558 vesicles and CTAB by UV-vis spectrum, stable fluorescence, transmission electron microscopy. The morphology of vesicles transformed with the change of surfactant concentration.

[Analysis of imatinib trough concentration at steady state in adjuvant therapy of patients with high risk gastrointestinal stromal tumor].

Objective: To explore the features of imatinib mesylate (IM) plasma concentration during adjuvant therapy and clinical factors associated with IM plasma concentration in patients with high risk gastrointestinal stromal tumors (GIST), and to determine whether IM plasma concentration <1100 µg/L influences the efficacy of adjuvant therapy. Methods: A retrospective case control study method was used. Case inclusion criteria: (1) complete resection of lesion and GIST confirmed by pathology; (2) high risk classified according to modified National Institutes of Health classification system (2008); (3) administration of IM 400 mg/d for at least 1 month; (4) not taking the medication likely affecting IM pharmacokinetic, such as rifampicin, dilantin, and carbamazepine, within 1 month before blood collection. Data of GIST patients who visited GIST Disease - Oriented Outpatient, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between January 2015 to December 2018 were retrospectively analyzed. After taking IM for 22-26 hours, 5 ml of peripheral venous blood was collected into EDTA anticoagulant tube. IM plasma concentration was detected by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Patients were divided into <1100 µg/L group and ≥1100 µg/L group according to plasma concentration. Linear regression was used to analyze the relevance between clinical features and IM plasma concentration. Parameters with normal distribution were analyzed by Pearson correlation coefficient, and parameters with non-normal distribution were analyzed by Spearman correlation. Kaplan-Meier survival curves and COX regression model were used for survival analysis. Results: Among the 85 patients enrolled in the study, 49 patients (57.6%) were male and 36 (42.4%) were female, with mean age of (51.9±11.0) years. The body mass index was (22.5±2.9) kg/m(2) and body surface area was (1.6±0.2) m(2). Thirty patients received gene test, including 23 patients with c-Kit exon 11 mutation, 4 with c-Kit exon 9 mutation, 1 with c-Kit exon 11 and 17 mutation and 2 without c-Kit or PDGFRA gene mutation. The mean IM plasma concentration was (1391.4±631.3) µg/L, and there were 32 patients with plasma concentration <1100 µg/L and 53 patients with plasma concentration ≥1100 µg/L. There were no statistically significant differences between the two groups in gender, age, body mass index, body surface area, hematological examination (white blood cells, albumin, alanine aminotransferase, aspartate aminotransferase and serum creatinine), tumor location, tumor size, mitotic counts, duration of adjuvant therapy and methods of operation (all P>0.05). Positive correlation between IM plasma concentration and serum creatinine was observed in linear regression analysis (r=0.297, P=0.007), but there were no correlations between IM plasma concentration and age (r=0.044, P=0.686), body mass index (r=0.066, P=0.547), body surface area (r=-0.010, P=0.924), white blood cells (r=-0.080, P=0.478), albumin (r=-0.065, P=0.563), alanine aminotransferase (r=0.114, P=0.308), aspartate aminotransferase (r=0.170, P=0.127) and duration of adjuvant therapy (ρ=0.060, P=0.586). There was no statistically significant difference in IM plasma concentration between patients with different genders (t=0.336, P=0.738) and patients with different surgical methods (F=0.888, P=0.451). Up to March 1, 2019. the median follow-up time was 30 (range 4-49) months. Tumor recurrence was detected in two patients with plasma concentration <1100 µg/L and two with plasma concentration ≥1100 µg/L. One recurrent patient with plasma concentration <1100 µg/L was detected to harbor c-Kit exon 11 and exon 17 mutations, and the other did not receive gene detection. Two recurrent patients with plasma concentration ≥1100 µg/L were both detected to harbor c-Kit exon 9 mutation. The 3-year relapse-free survival rate was 96.4% in the cohort, 96.2% in patients with plasma concentration <1100 µg/L, and 96.6% in patients with plasma concentration ≥1100 µg/L. No significant difference in relapse-free survival was observed between the two groups (P=0.204). Univariate Cox analysis showed that IM plasma concentration <1100 µg/L was not a risk factor for patients with high risk GIST (HR=0.238, 95% CI: 0.022-2.637, P=0.242). Conclusions: IM plasma concentration of adjuvant therapy in patients with high risk GIST varies with individual. Patients with higher level of serum creatinine are more likely to have a higher plasma concentration. A blood drug concentration standard of less than 1100 µg/L for advanced GIST patients may not influence the prognosis of patients with high risk GIST.

Exon skipping mutations in collagen VI are common and are predictive for severity and inheritance.

Mutations in the genes encoding collagen VI (COL6A1, COL6A2, and COL6A3) cause Bethlem myopathy (BM) and Ullrich congenital muscular dystrophy (UCMD), two related conditions of differing severity. BM is a relatively mild dominantly inherited disorder characterized by proximal weakness and distal joint contractures. UCMD was originally regarded as an exclusively autosomal recessive condition causing severe muscle weakness with proximal joint contractures and distal hyperlaxity. We and others have subsequently modified this model when we described UCMD patients with heterozygous in-frame deletions acting in a dominant-negative way. Here we report 10 unrelated patients with a UCMD clinical phenotype and de novo dominant negative heterozygous splice mutations in COL6A1, COL6A2, and COL6A3 and contrast our findings with four UCMD patients with recessively acting splice mutations and two BM patients with heterozygous splice mutations. We find that the location of the skipped exon relative to the molecular structure of the collagen chain strongly correlates with the clinical phenotype. Analysis by immunohistochemical staining of muscle biopsies and dermal fibroblast cultures, as well as immunoprecipitation to study protein biosynthesis and assembly, suggests different mechanisms each for exon skipping mutations underlying dominant UCMD, dominant BM, and recessive UCMD. We provide further evidence that de novo dominant mutations in severe UCMD occur relatively frequently in all three collagen VI chains and offer biochemical insight into genotype-phenotype correlations within the collagen VI-related disorders by showing that severity of the phenotype depends on the ability of mutant chains to be incorporated in the multimeric structure of collagen VI.

Complete exon-intron organization of the mouse fibulin-1 gene and its comparison with the human fibulin-1 gene.

Fibulin-1 is a 90 kDa calcium-binding protein present in the extracellular matrix and in the blood. Two major variants, C and D, differ in their C-termini as well as the ability to bind the basement membrane protein nidogen. Here we characterized genomic clones encoding the mouse fibulin-1 gene, which contains 18 exons spanning at least 75 kb of DNA. The two variants are generated by alternative splicing of exons in the 3' end. By searching the database we identified most of the exons encoding the human fibulin-1 gene and showed that its exon-intron organization is similar to that of the mouse gene.

Automated 3D segmentation of hippocampus based on active appearance model of brain MR images for the early diagnosis of Alzheimer's disease.

AIM: To investigate the hippocampal regional deformation modes by means of a novel method of automatic segmentation for discriminating between Alzheimer's disease (AD) and normal aging; and to further provide the effective evidence for the early diagnosis of AD. METHODS: Twenty AD patients and sixty healthy volunteers were included in this retrospective study. High-resolution structural volumetric images were obtained on a 3.0 T MR imaging system. Data were processed to create three-dimensional (3D) active appearance model (AAM) of hippocampus. Automatic recognition and 3D segmentation were carried out on both sides of the hippocampus in brain MR images of individuals with this model, and the hippocampal statistical shape model was established for AD group and control group. Student's t test was used to identify whether there was difference between AD group and control group in the hippocampal regional deformation detected by automatic segmentation, and to compare whether there was difference between the automated segmentation and the manual tracing for quantifying hippocampal volumes on left/right side of the same sex group of healthy volunteers and if there was genderwise difference. Pearson's Correlation test was employed to determine whether there was a correlation between automated segmentation and manual tracing for quantifying hippocampal volumes. RESULTS: No significant difference was detected between automated segmentation and manual tracing for quantifying hippocampal volumes on left/right side of the same sex group of healthy volunteers (P>0.05). Further there was no significant genderwise difference (P>0.05). A very strong positive correlation existed between both methods for quantifying hippocampal volumes (denoted R(2) near 1.0, P<0.001). Noticeable atrophy of bilateral hippocampal head was found among twenty patients with AD through statistical shape model compared with control group (P<0.05), especially on the left where inward-deformation was significantly found. CONCLUSION: This novel method of automated segmentation of the hippocampus based on AAM has been found to be reliable and accurate in our study, which may be an alternative to manual segmentation. The featured atrophy of hippocampal head can be regarded as an important biomarker for the early diagnosis of AD.

Reconstruction of mandibular symphyseal defects by an internal trifocal distractor: an experiment in dogs.

There have been few, if any, clinical reports on the use of an internal distractor to cure mandibular symphyseal defects. Compared with the currently-used external distractor, an internally implanted distractor may be more comfortable for patients, so we designed an internal trifocal distractor and tested its efficacy in animal models. We removed the mandibular symphyseal bones from mongrel dogs, producing a broken site 25-30 mm wide. Two transport discs, each roughly 10mm wide, were placed at the two ends of the broken bone, followed by assembly of the self-made internal trifocal distractor on the support of the two discs. A week after the procedure we initiated traction at a speed of 0.5mm/12h. The whole process lasted for 13-16 days until the two transfer discs had joined together at the centre of the mandible. Occlusal films were taken at the first, fourth, eighth, and twelfth week after traction. The animals were killed at the twelfth week, and the mandibles were examined histologically to assess the new bone formation. During the twelfth week after the initiation of traction, the physical continuity of the broken bones was restored. Histological results showed that the new bone formed in the traction area was abundant and had fully matured. We also found the formation of external rough callus, which indicated osteal concretion. Our results suggest that a well-designed internal trifocal distractor may be useful in the treatment of mandibular symphyseal defects.

[Study on the immunization status of children and risk factors in floating population].

In order to reveal immunization status of children in migrating families and its related risk factors, a study was conducted in Beijing, Inner Mongolia and Fujian from August to November, 1996. Results showed that the overall coverage with BCG, OPV, DPT and MV was 34.26% (95% CI: 29.75%-38.94%). Many factors influenced the immunization status of floating children which mainly consisted of a) regular searching for new coming floating children conducted by EPI staff, b) awareness of parents on the number of the total EPI-targeted vaccines, c) home as the place that the child was born, d) the length did the child's mother stay at the place, e) how well did the parents comply to the vaccination and f) The type of hospital was chosen when seeing a doctor.

[Economic burden of poliomyelitis].

Economic burden that caused by paralyzed and fatal poliomyelitis was described to provide basic data for evaluation. The study showed: the average costs for treatment, recovery and rehabilitation were 1612.6 yuan, 921.9 yuan and 1112.2 yuan per case, respectively. The total economic loss due to all new victims and handicapped cases of polio was 1.99 billion yuan in 1993 in China.

[Social burden caused by poliomyelitis].

Social burden caused by paralyzed poliomyelitis was analysed, using data from a Sampling Survey of Handicapped People in China, in 1987. The study results showed: 29.94% of 1707 handicapped polio victims aged 6 or over were not able to go to school; 44.23% of them could not take care of their daily life; 36.32% of them could not play out-door and do shopping; 2.58% of them could not normally communicate with others. 84.53% of the 1487 handicapped victims caused by polio aged 15 or over wholly or partly lost their working ability. We also compared handicapped people's employment rate and the status of marriage with non-handicapped people. Data showed that the employment rate and the number of married handicapped polio victims were remarkably lower then those non-handicapped people.

[Seroepidemiological surveillance and methodological study of pertussis, diphtheria and tetanus].

In this paper, we studied the diagnostic method of DPT using PA method developed by Japanese National Institute of Health (NIH) in 1993-1994, meanwhile we surveyed the antibody level of DPT. We collected 900 sera samples and 300 sera samples from Hebei province in 1993 and 1994. The result showed that PA method was simple, reliable and easy to observe, which can make up the shortages existed in PHA method. The antitoxin titers obtained by the PA method correlated well with the PHA method, while the latter was more sensitive. The antibody level of pertussis obtained through PA method did not correlate well with MA method. The result also showed that the antibody level of DPT was not high, with a 95.0% positive rate of pertussis. The rate of titers higher than 1:320 was 4.7% with GMT 1:121; the positive rate of diphtheria was 62.9% with GMT 0.0280 IU/ml, in 1993 the positive rate of tetanus was 70.5% with GMT 0.1410 IU/ml, in 1994 the positive rate was 75.7% with GMT 0.3281 IU/ml. The titers in urban areas were higher than in rural areas and a decrease was observed with the increase of age. The titers in males was similar to in females. According to this result, a routine immunization followed by booster program should be emphasized.

[Evaluation of acute flaccid paralysis (AFP) surveillance system in 1994].

Increased by 40.2% compared with 1993, 3142 acute flaccidparalysis cases were reported from AFP surveillance system in 1994. A total number of 1297 counties had AFP cases reported, accounting for 45.1% of total amount of counties. Out of 3142 AFP cases, 307 were confirmed, coming from 225 counties. For children under 15 years old, the reported AFP incidence was 1.1/10(5), non-polio AFP rate was 0.83/10(5), much higher than 0.37/10(5), in 1993. 48.5% of the total AFP cases had double stool specimen taken within 0-14 days after paralysis. 79% AFP cases had been investigated within 48 hours after being reported. Some of the major monitoring indicators of AFP surveillance system were close to the WHO criterion, but the percentage of AFP cases reported within 48 hours of careseeking and AFP cases with follow-up visit after 60-70 days of paralysis were very low. Furthermore, 77.9% of the confirmed cases were diagnosea clinically by residual paralysis, so did the 46.4% of discarded cases. It is in urgent need to increase the percentage of cases classified by laboratory test.

[Epidemiological analysis of poliomyelitis cases in 1994, China].

In 1994, the number of reported polio cases was 307 in China. This number was 53% lower than that in 1993, and was the lowest record ever in China. In 1994, the distribution of polio cases was highly dispersed and the number of outbreak was reduced obviously. Of 225 counties which reported polio cases in 1994, polio outbreak occured in 2 counties. Cases due to polio outbreak accounted for 4.2% of the total cases. The distribution of polio wild virus was narrow but the regional difference still existed. In the southeast, high risk region for polio consisting Guangdong, Hainan, Fujian three provinces, the number of polio cases decreased remarkably in 1994. On the contrary, in the southwest region which, consisting Guangxi, Yunnan, Guizhou, Sichuan four provinces, the number of polio cases increased obviously, accounted for approximately a half of the total number in 1994. During the coming 1 to 2 years, we should emphasis on these regions and provinces where high incidence was noticed in our polio eradication program. Those provinces should include Xinjiang, Fujian. Hubei three provinces that polio wild virus was identified in 1994, and Guangxi, Sichuan, Guizhou, Yunnan in the south west region as well. We should further increase Polio vaccination coverage of children especially floating and unplanned ones through strengthening routine immunization and carring out new national immunization day activity.

Cloning and sequence analysis of cDNAs encoding the alpha 1, alpha 2 and alpha 3 chains of mouse collagen VI.

cDNA clones encoding the alpha 1, alpha 2 and alpha 3 chains of mouse collagen VI have been isolated by screening cDNA libraries with the corresponding human probes. The composite cDNAs for the alpha 1, alpha 2, and alpha 3 chains are 2.5, 1.6 and 2.9 kb in size respectively. The alpha 1 and alpha 2 cDNAs encode the C-terminal portions of the chains as well as the entire 3'-untranslated regions, while the alpha 3 cDNAs encode a central segment of 959 amino acids flanking the triple-helical domain. The deduced amino acid sequences share 86-88% identity with the human counterparts and 67-73% identity with the chicken equivalents. Alignment of the deduced amino acid sequences of mouse, human and chicken collagens reveal that the key features of the protein, including the cysteine residues, imperfections in the Gly-Xaa-Xaa regions, Arg-Gly-Asp sequences and potential N-glycosylation sites, are mostly conserved.