Relationship between Changes of Pericyte Number and Wound Age during Repair of Skeletal Muscle Contusion in Mice.

ABSTRACT: Objective To observe the change pattern of pericyte number at different time periods after mice skeletal muscle contusion and discuss its role in wound age estimation. Methods A mice gastrocnemius muscle contusion model was established. The form and number changes of pericytes at 1, 3, 5, 7, 9, 14, and 28 d post-injury were detected by multiple immunofluorescence staining. Results Compared with the slender shape of pericytes in normal skeletal muscles, pericytes in the contusion area had increased volume, rounder form and a round nuclei. Part of pericytes were found to express satellite cell markers paired-box transcription factor （Pax7） or myoblast determination 1 （MyoD1）. The changes of pericyte number in skeletal muscles after contusion were time-dependant, and showed unimodal distribution with the extension of wound age. In the central contusion area, the number of pericytes peaked at 5 d post-injury while in the peripheral contusion area, the number of pericytes peaked at 5 d and 7 d post-injury. Conclusion The number of pericytes in contusion area varies time-dependently after skeletal muscle contusion in mice and might be a reference index for muscle wound age estimation, and is involved in the repair and regeneration of skeletal muscle injury.

Expression of Nrf2 in Different Cells after Human Cerebral Cortex Contusion.

ABSTRACT: Objective To observe the expression changes of nuclear factor-erythroid derived 2-related factors （Nrf2） in different cells at different time points after human cerebral cortex contusion, and to discuss its application in brain wound age estimation. Methods Thirty-six human brain tissues were selected, of which 6 were for control and 30 were cortical contusion at different time points post-injury, which were divided into 0-1 h, 3-6 h, 1-3 d, 5-7 d, and 10-14 d post-injury groups, with 6 cases in each group. Based on paraffin embedded sections, HE staining was used to observe the morphological changes post-injury, and double immunofluorescence staining was used to detect the expression of Nrf2 in neurons, astrocytes, and microglia. The number of positive cells was counted and statistical analysis was made. Results The number of neurons decreased 1-3 d post-injury. The expression of Nrf2 cells in neurons increased after injury, and the rate of positive cells peaked at 1-3 d post-injury. Glial cells were activated 1-3 d post-injury, and the activation peaked at 5-7 d post-injury. The cerebromalacia began to form at 10-14 d post-injury. Glial fibrillary acidic protein （GFAP） positive cells in mice increased gradually after injury and peaked at 5-7 d post-injury, while the proportion of Nrf2 in GFAP positive cells was relatively stable. After injury, ionized calcium-binding adapter molecule 1 （IBA1） positive cells increased and activated gradually. The expression proportion of Nrf2 in IBA1 positive cells increased gradually, reached its peak at 5-7 d post-injury, and then decreased. Conclusion The expression of Nrf2 in different cells involves in the biological function of different cells post-injury, and the dynamic expression of single cells has a time-dependent pattern. This may provide a new reference index for the wound age estimation of brain contusion in human.

Relationship between the Degeneration and Regeneration of Neuromuscular Junction and Wound Age during the Repair of Mouse Skeletal Muscle Contusion.

ABSTRACT: Objective To investigate the morphological changes in the degeneration and regeneration of neuromuscular junctions （NMJ） during the repair of mouse skeletal muscle contusion and discuss the correlation between the degeneration and regeneration of NMJ and wound age. Methods A total of 50 healthy adult male mice were randomly divided into 10 groups, including 9 experimental groups and 1 control group. Immunofluorescent staining was applied, and neurofilament was marked with neurofilament protein-H （NF-H）, presynaptic membrane was marked with synaptophysin （Syn）, presynaptic membrane was marked with acetylcholine receptor （AChR）. Morphological changes of NMJ regeneration at different time points after mouse skeletal muscle contusion were detected. Results The neurofilament and presynaptic membrane of NMJ at the junction of contusion zones began to degrade after contusion, and completed degradation at about 3 d post-injury. Then they gradually regenerated, roughly completing the regeneration at about 21 d and basically reaching the control group level. The ratio of presynaptic membrane quantity to presynaptic membrane quantity showed a trend of decreasing then rising and finally reaching the control level. Conclusion During the repair of mouse skeletal muscle contusion, the morphological changes and wound age of the NMJ at the junction of contusion zones have a close correlation, which is expected to be one of the biological indicators for forensic skeletal muscle wound age estimation.

Relationship between Expression Changes of CB2R and Wound Age of Brain Contusion in Mice.

ABSTRACT: Objective To investigate the expression of cannabinoid type 2 receptor （CB2R） at different time points after brain contusion and its relationship with wound age of mice. Methods A mouse brain contusion model was established with PCI3000 Precision Cortical Impactor. Expression changes of CB2R around the injured area were detected with immunohistochemical staining, immunofluorescent staining and Western blotting at different time points. Results Immunohistochemical staining results showed that only a few cells in the cerebral cortex of the sham operated group had CB2R positive expression. The ratio of CB2R positive cells gradually increased after injury and reached the peak twice at 12 h and 7 d post-injury, followed by a decrease to the normal level 28 d post-injury. The results of Western blotting were consistent with the immunohistochemical staining results. Immunofluorescent staining demonstrated that the changes of the ratio of CB2R positive cells in neurons, CB2R positive cells in monocytes and CB2R positive cells in astrocytes to the total cell number showed a single peak pattern, which peaked at 12 h, 1 d and 7 d post-injury, respectively. Conclusion The expression of CB2R after brain contusion in neurons, monocytes and astrocytes in mice suggests that it is likely to be involved in the regulation of the biological functions of those cells. The changes in CB2R are time-dependent, which suggests its potential applicability as a biological indicator for wound age estimation of brain contusion in forensic practice.

Validation of the traditional Chinese version of the Menopausal Rating Scale with WHOQOL-BREF.

Objective To assess the criterion validity, construct validity and test-retest reliability of the traditional Chinese language version of the Menopause Rating Scale (MRS-TC version). Methods This was an observational, cross-sectional study covering hospital and community samples of 317 women aged 39-62 years. Two questionnaires were administered, namely, the MRS-TC version, made up of 11 items in three dimensions, and the World Health Organization Quality of Life-BREF (WHOQOL-BREF). The intraclass correlation coefficient was used to examine the test-retest reliability of the questionnaire on two separate occasions, 2 weeks apart. The internal consistency was assessed with Cronbach's α. To evaluate criterion validity, the relationship between the individual items and dimension scores of both instruments was estimated. Pearson's correlation was used to assess convergent and discriminant validity; construct validity was evaluated by comparing the mean scores of menopausal and non-menopausal women for each of the MRS dimensions. Results The final questionnaire comprised 11 items in three dimensions. The intra-class correlation (ICC) for the test-retest reliability ranged from 0.83 to 0.93; values of Cronbach's α for psychological, somatic, and urogenital symptom domains were 0.88, 0.68, and 0.59, respectively. For the convergent and discriminant validity, the correlations between the individual questionnaire and the WHOQOL-BREF were significant; those with the MRS dimensions were significantly negatively associated for the physical, psychological, social and environmental domains. Conclusion The MRS-TC version using the traditional Chinese language is a reliable and valid questionnaire for assessing menopausal symptoms and global quality of life in climacteric women.

The impact of plasma Epstein-Barr virus DNA and fibrinogen on nasopharyngeal carcinoma prognosis: an observational study.

BACKGROUND: The impact of combining plasma fibrinogen levels with Epstein-Barr Virus DNA (EBV DNA) levels on the prognosis for patients with nasopharyngeal carcinoma (NPC) was evaluated. METHODS: In this observational study, 2563 patients with non-metastatic NPC were evaluated for the effects of circulating plasma fibrinogen and EBV DNA levels on disease-free survival (DFS), distant metastasis-free survival (DMFS), and overall survival (OS). RESULTS: Compared with the bottom biomarker tertiles, TNM stage-adjusted hazard ratios (HR, 95% confidence intervals (CIs)) for predicting DFS in fibrinogen tertiles 2 to 3 were 1.26 (1.00 to 1.60) and 1.81 (1.45 to 2.26), respectively; HR for EBV DNA tertiles 2 to 3 were 1.49 (1.12 to 1.98) and 4.24 (3.27 to 5.49), respectively. After additional adjustment for established risk factors, both biomarkers were still associated (P for trend <0.001) with reduced DFS (HR: 1.79, 95% CI, 1.43 to 2.25 for top fibrinogen tertiles; HR: 4.04, 95% CI: 3.10 to 5.27 for top EBV DNA tertiles compared with the bottom tertiles). For patients with advanced-stage disease, those with high fibrinogen levels (3.34 g l(-1)) presented with worse DFS, regardless of EBV DNA 4000 or <4000 copies ml(-1) subgroup. Similar findings were observed for DMFS and OS. CONCLUSIONS: Circulating fibrinogen and EBV DNA significantly correlate with NPC patients survival. Combined fibrinogen and EBV DNA data lead to improved prognostic prediction in advanced-stage disease.

Role of 15-F2t-isoprostane in intestinal injury induced by intestinal ischemia/reperfusion in rats.

15-F2t-isoprostane is not only a specific marker of lipid peroxidation but also demonstrated to have potent bioactivities and can exert deleterious effects via activating thromboxane A2 receptor (TxA2r). We already demonstrated that lipid peroxidation represents a mechanism of intestinal ischemia/reperfusion (I/R) injury. But no studies have focused on 15-F2t-isoprostane production and its biological actions on postischemic intestine during intestinal I/R. This study was carried to investigate whether the mechanism of endogenous 15-F2t-isoprostane action is involved in the pathogenesis of intestinal I/R and administration of synthetic 15-F2t-isoprostane could exacerbate intestinal insult after intestinal I/R in vivo and in vitro. In comparison with that of the sham control, we reported that endogenous 15-F2t-isoprostane was liberated following intestinal I/R injury in rats, and using the TxA2r antagonist SQ29548 resulted in significant intestinal protection, evidenced by reduced lipid peroxidation, inflammation, and alleviated intestinal mucosal microvascular vasoconstriction. Further research found that in vivo administration of synthetic 15-F2t-isoprostane exacerbated intestinal I/R injury by disturbing microvascular perfusion and accumulating anaerobic metabolism. Meanwhile, 15-F2t-isoprostane did not change Hypoxia/Reoxygenation-induced IEC-6 cell viability but aggravated HUVECs cell death in vitro. Collectively, our results showed that locally produced 15-F2t-isoprostane was in proportion to the severity of oxidative stress-induced intestinal injury and its detrimental effects can be attenuated through TxA2r inactivation. Exogenous 15-F2t-isoprostane exacerbated intestinal I/R injury, which may be contributable to its biological actions on endothelium, rather than intestinal epithelium.

Molecular cloning and functional characterization of a mouse ccl6 analog gene in the rat.

Suppression subtractive hybridization was used to analyze differential expression of genes in rat peritoneal macrophages after granulocyte macrophage colony-stimulating factor treatment. We identified and cloned the mouse C10 analog gene in the rat, and named it as ccl6. The full-length cDNA of rat ccl6 was 467 bp, which contains a single-open reading frame and encodes 116 amino acid residues. Compared with other C-C chemokines, the rat ccl6 gene had an unusual four-exon genome structure instead of the typical three exons, it had the highest homology with murine ccl6. The rat ccl6 gene was localized on chromosome 10, where most of the C-C chemokine superfamily members are located. The recombinant rat C-C chemokine ligand 6 (CCL6) protein was expressed by the pGEX4T-1 plasmid in Escherichia coli BL21. The purified recombinant protein had bioactivity similar to that of mouse CCL6, which is a chemoattractant for macrophages and lymphocytes, but not for neutrophils.

A Bayesian spatial multimarker genetic random-effect model for fine-scale mapping.

Multiple markers in linkage disequilibrium (LD) are usually used to localize the disease gene location. These markers may contribute to the disease etiology simultaneously. In contrast to the single-locus tests, we propose a genetic random effects model that accounts for the dependence between loci via their spatial structures. In this model, the locus-specific random effects measure not only the genetic disease risk, but also the correlations between markers. In other words, the model incorporates this relation in both mean and covariance structures, and the variance components play important roles. We consider two different settings for the spatial relations. The first is our proposal, relative distance function (RDF), which is intuitive in the sense that markers nearby are likely to correlate with each other. The second setting is a common exponential decay function (EDF). Under each setting, the inference of the genetic parameters is fully Bayesian with Markov chain Monte Carlo (MCMC) sampling. We demonstrate the validity and the utility of the proposed approach with two real datasets and simulation studies. The analyses show that the proposed model with either one of two spatial correlations performs better as compared with the single locus analysis. In addition, under the RDF model, a more precise estimate for the disease locus can be obtained even when the candidate markers are fairly dense. In all simulations, the inference under the true model provides unbiased estimates of the genetic parameters, and the model with the spatial correlation structure does lead to greater confidence interval coverage probabilities.

A grid-search algorithm for optimal allocation of sample size in two-stage association studies.

Multiple testing occurs commonly in genome-wide association studies with dense SNPs map. With numerous SNPs, not only the genotyping cost and time increase dramatically, many family wise error rate (FWER) controlling methods may fail for being too conservative and of less power when detecting SNPs associated with disease is of interest. Recently, several powerful two-stage strategies for multiple testing have received great attention. In this paper, we propose a grid-search algorithm for an optimal design of sample size allocation for these two-stage procedures. Two types of constraints are considered, one is the fixed overall cost and the other is the limited sample size. With the proposed optimal allocation of sample size, bearable false-positive results and larger power can be achieved to meet the limitations. The simulations indicate, as a general rule, allocating at least 80% of the total cost in stage one provides maximum power, as opposed to other methods. If per-genotyping cost in stage two differs from that in stage one, downward proportion of the total cost in earlier stage maintains good power. For limited total sample size, evaluating all the markers on 55% of the subjects in the first stage provides the maximum power while the cost reduction is approximately 43%.

[Enteral nutrition in the severely head injured patients].

This study evaluated the effect of modified element diet as early nutritional support in 20 severe head-injured patients. The study group (20 cases) was fed gastrically with the element diet of dextrin as glycogen by a nasal feeding tube, and the control group (20 cases) was nutritional supported with common method. The enteral nutrition (EN) was started within 5.0 +/- 2. 1 days post injury for study group versus 9.1 #+/- 4.6 days for control group (P < 0.05). The serum albumin levels of the study group rose in the third week post injury, the data of the forth week was slightly higher than that of the first week. In control group, the serum albumin levels reduced continuously within four weeks post injury. The serum glucose levels of the study group was significantly lower than that of the control group on the fifth day post injury (P < 0.05). 45% patients in the study group had diarrhea. This study showed that the modified element diet can be helpful for the early enteral nutrition and beneficial to improve the nutritional condition of the severe head-injured patients.

Plasma luteinizing hormone and progesterone concentrations after prostaglandin F2 alpha in Taiwan yellow cows (Bos indicus).

A luteolytic dose (500 micrograms) of cloprostenol was given four days before expected estrus to five mature cyclic cows of native Taiwan Yellow cattle. Blood samples were collected at hourly intervals for 60 hours and then twice daily (0800 h and 1600 h) until onset of the following estrus or Day 29 after prostaglandin (PG) injection. Plasma progesterone levels were quite low in the follicular phase and elevated over an 11-13 day period during the luteal phase for every animal. Two cows showed peak luteinizing hormone (LH) levels of 19.97 and 23.77 ng/ml of plasma at 120 and 127 h after PG injection. One cow did not show peak LH level during the 60-h sampling period, but had its highest concentration of 18.02 ng/ml at 0800 h on Day 8. Two cows showed no detectable peak LH during the 60-h sampling period or on subsequent days. But these two cows had increased concentrations of progesterone beginning on Day 6 and Day 12. All animals came back into estrus 23-28 days after PG treatment.

Seasonal variations in plasma progesterone concentrations in cyclic Taiwan yellow cattle (Bos indicus)

Plasma concentrations of progesterone were measured by radioimmunoassay in plasma samples obtained from 11 mature cyclic cows of native Taiwan Yellow cattle during the warm (March-April) and cool (November-December) seasons. Progesterone levels began to increase after Day 4 (Day 0 = the day of estrus) and reached maximum values on Day 10 and Day 14 for cool and warm seasons, respectively. Progesterone concentrations in plasma during the estrous cycle were significantly higher (p < 0.001) in the cool season (2.66 +/- 0.19 ng/ml) than in the warm season (1.98 +/- 0.15 ng/ml).

[Study on omeprazole 20 mg twice weekly in prevention of duodenal ulcer relapse].

The aim of this study is to propose a regimen of omeprazole 20 mg twice weekly for the prevention of duodenal ulcer (DU) relapse. 257 endoscopically verified DU patients entered this study. They were divided into four groups: (1) Group A, 68 patients stopped treatment after healing of DU during treatment of cimetidine 800 mg at night. (2) Group B, 72 patients healed DU after treatment of omeprazole 20 daily. Then anti-ulcer drug was withdrawn. (3) Group C, after healing of DU, 54 patients received a maintenance dosage of cimetidine 400 at night for up to 1 year. (4) Group D, after healing of DU, 63 patients received omeprazole 20 mg twice weekly (every Monday and Thursday) for up to 1 year. All the patients were followed in out-patients department for assessment of symptomatic response and side effect. Endoscopic examination were performed in those patients when any symptom related to the upper gastrointestinal tract appeared. The cumulative recurrence rates of Group A to Group D at 6 months were 44.1%, 23.6%, 22.2% and 4.8%, and that at 12 months were 76.5%, 36.1%, 27.8% and 6.3% respectively. The relapse rate of patients with omeprazole 20 mg twice a week was significantly lower than that of other groups at either 6 or 12 months. In patients with maintenance therapy of cimetidine, DU recurred more rapidly among smokers than nonsmokers, but there is no significant difference between smokers and nonsmokers receiving maintenance therapy of omeprazole.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effect of CCK receptor antagonists on plasma CCK bioassay].

Fasting and postprandial plasma CCK levels of 102 normal subjects were measured by bioassay with dispersed rat pancreatic acini. The reference values ranged from 0 to 4.2 pmol/L (CCK-8 equivalents) for fasting and from 1.1 to 13.5 pmol/L for postprandial state. There was no significant difference between male and female, or in different age groups. The effects of CCK receptor antagonists of 3 different categories on CCK bioactivity in plasma measured by the bioassay were investigated. L 364,718 (5 nmol/L), proglumide (1.0 mmol/L), or Bt2-cGMP (0.1 mmol/L) was either extracted by SEP-PAK C18 cartridges together with human plasma containing 8 pmol/L of CCK-8, or added into the plasma extracts before the assay. The CCK bioactivity was inhibited by all of the 3 CCK antagonists. The action of L364,718 could be eliminated by the procedure of plasma extraction, but not of proglumide or Bt2-cGMP. It was suggested that CCK bioassay can be used even if L364,718 was administered. However, CCK cannot be measured accurately if there are proglumide or Bt2-cGMP in the plasma.

[Protective effect of colloidal bismuth subcitrate on gastric mucosal lesion induced by aspirin].

This study was designed to investigate the protective effect of colloidal bismuth subcitrate (CBS) on aspirin-induced gastric mucosal lesion 20 patients with arthralgia were allocated into this study. All of them were free of gastrointestinal symptom and their gastric mucosa were nearly normal under gastroendoscopic observation. The first group of 10 patients received orally aspirin 1.5 qid for 4 days. The second group also of 10 patients was treated with CBS 120 mg qid and after 2 days they received aspirin and CBS simultaneously in the dosage mentioned above for another 4 days. Before and after treatment, the patients of both groups were examined endoscopically. After treatment, the mucosal inflammation was much less in the second group than that in the first group. The prostaglandin E2 concentration of antral mucosa in the first group was reduced significantly after administration of aspirin alone, while that in the second group was slightly increased after a combined treatment of aspirin and CBS. The results demonstrate that CBS is an effective agent in prevention of gastric lesion induced by aspirin and prostaglandin E may be involved in this mucosal protective effect.