Submucosal injection of pharyngeal ostium of Eustachian tube for diagnosis of patulous Eustachian tube / 临床耳鼻咽喉头颈外科杂志

To evaluate the clinical value of submucosal injection of pharyngeal ostium of Eustachian tube in diagnosing patulous Eustachian tube(PET).Twenty-six patients(32 sides),whose the symptoms were consistent with PET,were enrolled from March 2014 to May 2016.The symptoms and signs of all patients were evaluated after submucosal injection of saline into the Eustachian tube.Immediately after submucosal injection of saline into the Eustachian tube,the symptoms and signs disappeared in 24 cases(29 sides),and improved in 2 cases(3 sides).The resolution and/or improvement of symptoms and signs lasted for less than 24 hours in 12 patients,for more than 24 hours in 9 patients,and for more than 48 hours in 4 patients.No adverse reactions were observed.Submucosal injection may be a simple and practical method for auxiliary diagnosis of PET,and may be used in preoperative evaluation of Eustachian tuboplasty.

Histological structure of the trabecular meshwork in the eyeball: challenging the traditional concept and preliminary findings in rabbits, rats and mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To verify that the trabecular meshwork (TM) in the wall of the eyeball consists of smooth muscle fibers instead of collagen fibers or endothelial cells.</p><p><b>METHODS</b>Eighteen fresh eyeballs from 3 rabbits, 3 SD rats and 3 mice were sectioned along the sagittal plane and sliced after paraffin embedding for HE staining, VG staining, Masson staining, α-SMA immunohistochemistry or CD31 immunohistochemistry. These slices were observed under microscope and the structure of the TM was compared with those of scleral collagen fibers, ciliary muscles and endothelial cells.</p><p><b>RESULTS</b>HE staining of the eyeball slices from the 3 animal species resulted in purplish red staining of the TM, which was highly consistent with ciliary muscle fibers. The cell?like structures on the surface of the TM were not clearly outlined, with flat nuclei showing a dark purple staining; these structures did not show obvious boundaries from the TM. Ciliary muscle fibers, which were smooth muscle cells in nature, were aligned in bundles in various directions. The longitudinally sectioned cells were flat and contained purplish cytoplasm and highly flattened nuclei. Scleral collagen fibers were stained dark red with a few fibroblasts sandwiched among them. The long axis of the fibroblasts was in parallel with that of the collagen fibers. The outline of the fibroblast was not clear and the nucleus was flat in dark blue. The vascular endothelial cells presented with different morphologies and contained light purplish cytoplasm and dark nuclei, protruding into the vascular cavity. VG staining of the TM revealed a pale red filamentous structure, and the collagen fibers were stained bright red. Masson staining of the TM showed a reticular structure consisting mainly of dark red fibers intermingled with thin green fibers. Scleral collagen fibers presented with a cord?like green wavy structure. The endothelial cells were green and flat, while the ciliary smooth muscle fibers were purple. In immunohistochemistry for α?SMA, the TM and the ciliary smooth muscle fibers showed a strong positivity in the cytoplasm, while the scleral collagen fibers and vascular endothelial cells showed negative staining; immunohistochemistry for CD31 showed no obvious positive staining in the TM, collagen fibers or ciliary smooth muscle cells from all the animals in spite of slight differences among them.</p><p><b>CONCLUSION</b>The TM consists mainly of smooth muscle fibers with a thin layer of peripheral endomysium without endothelial cells.</p>

The generation of the endothelial specific cdc42-deficient mice and the effect of cdc42 deletion on the angiogenesis and embryonic development / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>High microvascular permeability plays an essential role in pathological process of multiple diseases such as septic shock, acute lung injury and acute respiratory distress syndrome, and burns. Inhibiting hyperpermeability is significant for controlling these conditions. Cdc42, as a main member of the small Rho GTPase family, plays a critical role in controlling and regulating the endothelial junctional permeability. We aimed to generate and identify endothelial specific cdc42-deficient mice by the Cre/loxp recombination approach, for examination in an animal model of the contribution of the cdc42 gene in the microvascular barrier function.</p><p><b>METHODS</b>We crossed cdc42(Flox/Flox) mice with mice expressing endothelial cell-specific Cre recombinase, and the offspring with the genotype cdc42(Flox/+)Tie2Cre(+/-) were back-crossed with the cdc42(Flox/Flox) mice. The cdc42(Flox/Flox)Tie2Cre(+/-) mice in the F2 generation were the target mice. If the cdc42 deficient mice did not survive, we would observe the cdc42 deficient mice embryos, and compare them with wild-type mice embryos.</p><p><b>RESULTS</b>Cdc42(flox/+)Cre(+/-) mice were mated with the cdc42(Flox/Flox) mice and among the living offspring there were no cdc42(Flox/Flox)Cre(+/-) target mice. We found the endothelial special cdc42 deficient embryos at the E7.5-E16.5 stage. We observed that cdc42 deficient embryos were much smaller, had fewer vessels and were a little more swollen compared with the wild-type embryos.</p><p><b>CONCLUSIONS</b>Endothelial specific knockout of cdc42 caused embryonic lethality and the mice did not survive to birth. The target embryos were much smaller, had fewer vessels and were a little more swollen compared with the wild-type embryos. These results demonstrated that the cdc42 plays an important role in development of embryos and in development of microvessels as well as microvascular permeability.</p>

A two-step protocol for induction of rat adipose tissue-derived stem cells into neuron-like cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differentiation potential of rat adipose tissue-derived cells (ADSCs) into neuron-like cells in vitro using a two-step induction protocol.</p><p><b>METHODS</b>ADSCs isolated from the epididymal fat pads in male SD rats by means of differential attachment were cultured in vitro and subjected to adipogenic induction. After flow cytometric identification of the cell surface antigens CD106, CD11b, CD45, CD49d, CD90 and CD29, the third-passage ADSCs were induced to transdifferentiate into neural stem cell (NSC)-like cells in DMEM/F12 medium containing 10 ng/ml basic fibroblast growth factor (bFGF), 20 ng/ml epidermal growth factor (EGF) and 2% B27. The resultant NSC-like cells were then induced to differentiate into neuron-like cells in the neurobasal medium containing 10 ng/ml brain-derived neurotrophic factor (BDNF), 10 ng/ml glial cell line-derived neurotrophic factor (GDNF) and 1 µmol/L retinoic acid (RA). Immunocytochemistry was employed to identify the expression of the cell surface markers nestin, MAP2 and NeuN.</p><p><b>RESULTS</b>The isolated ADSCs were positive for CD90 and CD29, and oil red O staining of the induced adipose-like cells yielded positive results. The third-passage ADSCs induced for 7 days aggregated as floating cell spheres positive for NSC surface antigen nestin. Further induction in neurobasal medium for 4 h resulted in adhesion of the cell spheres and the formation of cell processes extending from some peripheral cells, suggesting a morphological resemblance to neurons. Most of the cells showed positivity for MAP2 and NeuN.</p><p><b>CONCLUSION</b>ADSCs can be induced to differentiate into neuron-like cells in vitro under appropriate conditions.</p>

The comparison between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice on lung tissue pathological change and vasopermeability in acute lung injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the change of lung tissue and vasopermeability between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice in acute lung injury.</p><p><b>METHODS</b>The mice with vascular endothelial cell-specific expression of cre recombinase were crossed with cdc42(flox/flox) mice. The cdc42(flox/+)Cre(+/-) F1 offspring mice were crossed back with cdc42(flox/flox) mice, resulting in the F2 generation mice with three genotypes, namely cdc42(flox/+)Cre(+/-), cdc42(flox/flox)Cre(-/-) and cdc42(flox/+)Cre(+/-). The heterozygous mice with cdc42(flox/+)Cre(+/-) genotype were selected as the model mice, with the other two genotype groups as the control. After intratracheal instillation of 2 mg/kg LPS to induce acute lung injury, the mice were sacrificed to examine the lung pathologies, lung wet/dry ratio and lung microvascular permeability.</p><p><b>RESULTS</b>The heterozygous mice with cdc42 gene knockout (cdc42(flox/+)Cre(+/-)) showed no significant differences from the two control groups in the lung pathological score, lung wet/dry ratio or the lung microvascular permeability coefficient.</p><p><b>CONCLUSION</b>There were no significant difference on lung tissue and vasopermeability between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice.</p>

Forward genetic screening for zebrafish mutants defective in erythropoiesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen and identify zebrafish mutants with erythropoiesis defects by N-ethyl-N-nitrosourea (ENU) mutagenesis and large-scale forward genetic screening using beta e 1 as the marker.</p><p><b>METHODS</b>The chemical mutagen ENU was used to treat healthy wild-type male fish (AB strain, F0). The surviving ENU-treated fish were mated with wild-type female fish to generate F1, and further F2 family was generated by F1 family intercross. The adult F2 fish were intercrossed within each F2 family and the resulting F3 embryos from each crossing were subjected to whole mount in situ hybridization (WISH) with the beta e 1 probe. Mutagenesis was performed by treating the male zebrafish with ENU to induce mutations in pre-meiotic germ cells to generate the founders, which were outcrossed to obtained the F1 fish. The F1 fish from different founders were mated to generate the F2 families. F3 embryos from the sibling cross in the F2 family were examined by whole mount in situ hybridization using beta e 1-globin probe. The putative mutants were then characterized with different hematopoiesis markers.</p><p><b>RESULTS AND CONCLUSION</b>We identified 4 beta e 1-deficient mutants with erythropoiesis defects, including two with specific erythiod lineage defects and two with concurrent lymphopoiesis defects.</p>

Forward genetic screening for zebrafish mutants defective in myelopoiesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify zebrafish mutants with myelopoiesis defects by ENU mutagenesis and large-scale forward genetic screening.</p><p><b>METHODS</b>Male zebrafish were mutagenized with N-ethyl N-nitrosourea to induce mutations in the spermatogonial cells to generate the founders, which were outcrossed with AB to raise F1 fish. The F1 fish from different founders were mated to generate the F2 families. The F3 embryos from F2 sibling crosses were screened by Sudan black B staining and neutral red staining.</p><p><b>RESULTS</b>A total of 350 F2 families from F1 sibling crosses were screened, and 1424 F2 crosses were analyzed. Six mutations were identified resulting in abnormal Sudan black B staining and neutral red staining, indicating the involvement of neutrophil deficiency or macrophage abnormalities.</p><p><b>CONCLUSION</b>It is simple and cheap to induce and screen myelopoiesis deficiency in zebrafish by ENU chemical mutagenesis and Sudan black B staining and neutral red staining. These mutants shed light on the identification of the genes important to myelopoiesis in zebrafish.</p>

Induced differentiation of rat adipose-derived stem cells into Schwann-like cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the differentiation potential of rat adipose-derived stem cells (ADSCs) into Schwann-like cells in vitro.</p><p><b>METHODS</b>ADSCs isolated from adult SD rats were cultured in vitro and identified with the cell surface antigens CD44, CD49d and CD106 by immunocytochemistry. The ADSCs of the sixth to eighth passages were inoculated in polylysine-coated culture plate and cultured for 12 days in DMEM/F12 culture medium containing 10% fetal bovine serum, 5 ng/ml platelet-derived growth factor, 10 ng/ml basic fibroblast growth factor, 14 micromol/L Forskolin and 200 ng/ml Heregulin to induce their differentiation in vitro. Immunocytochemistry was performed to identify the expression of the cell surface markers nestin, glial fibrillary acidic protein (GFAP), S-100, and P75.</p><p><b>RESULTS</b>The isolated and purified ADSCs were positive for CD44 and CD49d expressions but negative for CD106. After 12 days of culture in the conditional culture medium, most of the cells showed positive expressions of GFAP, S-100, and P75, the specific protein markers of Schwann cells.</p><p><b>CONCLUSION</b>Adult rat ADSCs are confirmed to have potentials of neuroglial differentiation and capable of differentiating into Schwann-like cells in vitro.</p>

Model for end-stage liver disease-sodium predicts prognosis in patients with chronic severe hepatitis B / 中华外科杂志

<p><b>OBJECTIVES</b>To study the practical use of the serum sodium incorporated model for end-stage liver disease (MELD-Na) on clinic and to assess its validity by the concordance-statistic in predicting the prognosis of the patients with chronic severe hepatitis B.</p><p><b>METHODS</b>Adult patients with a diagnosis of chronic severe hepatitis B between January 2007 and December 2007 in a single center were analyzed. The serum sodium, MELD, MELD-Na, and Delta MELD-Na (Delta MELD=MELD score at 14 days after medical treatment-MELD score at admission) scores of 426 patients with chronic severe hepatitis B were calculated. The 3-month mortality in patients was measured, and the validity of the models was determined by means of the concordance-statistic.</p><p><b>RESULTS</b>The area under the receiver-operating characteristic curves of Na, MELD and MELD-Na for the occurrence of death in 3 month were 0.718, 0.875 and 0.922. The 3-month mortality of the MELD-Na scores group <25, 25-30, >30-35, >35- <40 and > or = 40 were 2.0%, 5.4%, 35.4%, 53.8% and 86.9% respectively. There was a significant difference of 3-month mortality between the five groups (P<0.05). The 3-month mortality of Delta MELD-Na> 0 group was 65.9%, and the Delta MELD-Na < or = 0 group was 15.8%. There was a significant difference of 3-month mortality between the two groups (P<0.05).</p><p><b>CONCLUSIONS</b>MELD-Na score is a valid model to predict 3-month mortality in patients with chronic severe hepatitis B. Delta MELD-Na is clinically useful parameters for predicting the therapeutic effect of chronic severe hepatitis B.</p>

Identification of factors associated with YMDD mutation in patients with chronic hepatitis B receiving lamivudine treatment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To identify factors associated with YMDD mutation in patients with chronic hepatitis B before and after lamivudine treatment in Zunyi region.</p><p><b>METHODS</b>53 patients with chronic hepatitis B were enrolled in this study, HBV DNA,HBV markers, ALT, AST, TBil, albumin in the serum were examined at 0, 3, 6, 12, 18 and 24 months after lamivudine treatment. HBV genotype and YMDD mutation were determined by sequencing before lamivudine treatment. YMDD mutation was checked again if serum HBV DNA rebound to more than 1 x 10(4) copies/ml after the initial decrease.</p><p><b>RESULTS</b>HBV genotype in Zunyi region is constitute of B, C and B+C genotype. YMDD mutation occurred in 18 cases after lamivudine treatment, the rate of YMDD mutation was 15.1%, and 34.0% after 1 year and 2 years treatment. There are four types of mutation: rtL180M/M204V, rtL180M/M204I, rtM204I, rtL180M. rtM204V mutation in C gene was always accompanied by rtL180M mutation (100%). The rate of rtL180M/M204V mutation in genotype C group was significantly higher than that in genotype B group (77.8% to 25.0%), the same was true for the rtL180M/ M204I mutation (22.2% to 12.5%). There was no point mutation in genotype C group. The point mutation of rtM204I and rtL180M appeared only in genotype B group. Gender, nation, family history of hepatitis B and HBeAg were not associated with YMDD mutation (P more than 0.05), while the mutation rate was associated with the disease course and severity of disease. YMDD mutation did not occur in patients with low HBV DNA level (less than 10(5) copies/ml).</p><p><b>CONCLUSION</b>YMDD mutation after lamivudine therapy is associated with HBV genotype and P gene mutation type, and prolonged treatment increases the the mutation rate. In order to reduce the incidence of YMDD mutation, patients with shorter disease course, lower HBV DNA level, more serious liver damage should be treated with lamivudine.</p>

Separation and purification processes of alkaloids from Lotus plumule with macroporous adsorption resin / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the optimal separation of the alkaloids from Lotus plumule by selecting appropriate macroporous adsorption resins.</p><p><b>METHOD</b>To evaluated the separating efficiency by measuring the adsorption ratio, eluting ratio and the concentration of neferine.</p><p><b>RESULT</b>The LSA -5B macroporous adsorption resin prolided the optimal separating efficiency. The best adsorption capacity was achieved with the following conditions: the concentration of extact liquid was 0. 125 g x mL(-1) (equivalence to raw material), then the resin was washed by water to remove impurity and alkaloids were desorbed by 50% ethanol.</p><p><b>CONCLUSION</b>This method is simple, feasible and suitable for industry production.</p>

Biological dressing with human hair keratin-collagen sponge-poly 2-hydroxyethyl methacrylate composite promotes burn wound healing in SD rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a composite material containing human hair keratin (HHK), collagen sponge (inner layer) and poly 2-hydroxyethyl methacrylate (PHEMA) film that allows sustained release of polydatin and test its effect as a biological dressing in promoting burn wound healing in SD rats.</p><p><b>METHODS</b>Three HHK materials with fast, moderate, and low degradation rates were mixed at the ratio of 4:3:3 to prepare a reticular structure, which was processed into a composite material with bovine tendon-derived collagen sponge, and further complexed with HEMA film containing PD prepared by polymerization. Degree II burn wound was induced in SD rats by scalding and within postburn day 2-5, the wounds were cleansed and covered with the composite material or with glutaraldehyde-treated porcine skin (positive control). At week 1, 2, 4, 6 and 8 following wound dressing, 6 full-thickness skin samples were harvested from the wounds for histological observation and immunohistochemical detection of collagen and elastic fibers, and the wound healing time and healing rate were recorded.</p><p><b>RESULTS</b>The prepared collagen sponge film was transparent and porous (50-300 microm in diameter) and allowed sustained PD release into normal saline within 48 h. Compared with the porcine skin, the composite material reduced exudation and maintained ideal moisture of the wound, and significantly shortened the wound healing time (P=0.000). On day 7, 14, and 21 following dressing, the composite material and porcine skin significantly increased the wound healing rate as compared with the negative control group (P=0.000), and on day 14, the composite achieved significantly greater healing rate than the porcine skin (P<0.05).</p><p><b>CONCLUSION</b>HHK-collagen sponge-PHEMA/PD composite as a dressing material promotes burn wound healing in rats by allowing in vivo construction of tissue engineered epidermis. PHEMA is feasible for sustained drug delivery in this composite.</p>

Preliminary study of chicken calamus conduit as a scaffold material for tissue engineering / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the unique structural features of chicken calamus keratin (CCK) conduit as a candidate scaffold material for tissue engineering and its in vivo degradation and histocompatibility after its implantation into living tissues.</p><p><b>METHODS</b>Chicken calami were taken from healthy chickens and treated through sequential, controllable physical and biochemical procedures for preparation of three types of CCK conduits, namely CCK-I (mildly treated), CCK-II (moderately treated) and CCK-III (intensely treated). Light microscopy (LM) and scanning electron microscopy (SEM) were performed for morphological observation. Each of these three types of CCK pieces (experimental group) and the untreated ones (control group) was implanted into the dorsal muscular tissue on both sides of SD rats, respectively. Routine tissue sectioning and HE stain were performed to identify the morphological changes under light microscope. Each of the CCK threads (experimental group) and the untreated chicken calamus threads (control group) was also grafted within the sciatic nerve bundles of SD rats, respectively.</p><p><b>RESULTS</b>The wall of the chicken calamus was composed of 4 compact parts from inside to outside on cross sections, namely the innermost basophilic homogenous coarse line, 3-5 layers of acidophilic corneum, 60-100 layers of circular keratin tracts containing massive pigment granules, and 10-20 outmost layers of keratin tracts with only a few pigment granules. The three-dimensional surface features of chicken calamus identified by SEM, as compared with untreated chicken calamus, was characterized by loose arrangement containing horizontal and vertical keratins with obvious pores of different sizes and depths on its surface. At 8 weeks after implantation into the muscular tissue in experimental groups, the CCK grafts were degraded into thin filaments or/and dispersed pieces and fine granules with the appearance of blood vessels, which facilitated the absorption of the degradation products; at 12 weeks, the grafts were markedly degraded into tiny fragments. In the control group, in contrast, the grafts remains intact throughout the experiment. After implantation of the material into the nerve bundles, similar cell infiltration and tissue responses to the grafts were observed as compared to those occur in intramuscular grafting. The degradation products did not seem to cause nerve tissue degeneration or necrosis.</p><p><b>CONCLUSIONS</b>Fresh chicken calamus is a natural tube composed of multi-layered compact keratin tracts with pigment granules and small amount of matrix, and is non-absorbable in vivo, and therefore does not favor the purpose for use directly as a candidate biological scaffold. After proper treatment, the chicken calamus becomes loosely arranged porous material, and can be degraded and absorbed in vivo without resulting in tissue degradation or necrosis, suggesting its potential for applications in tissue engineering.</p>

Toxicity evaluation of chicken calamus keratin conduit as a tissue-engineering scaffold biomaterial / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the toxicity of chicken calamus keratin (CCK) conduit as a tissue-engineered scaffold material.</p><p><b>METHODS</b>The chemical composition of the leaching solution of CCK was determined by means of ultraviolet spectrometry, and the toxic effects of the solution was evaluated by skin sensitization test in rats, intracutaneous stimulation test in rabbits, acute systemic toxicity test in mice, and cytotoxicity test in L929 cells.</p><p><b>RESULTS</b>The leaching solution of CCK consisted mainly of middle-molecular-weight peptides with a small quantity of macromolecular proteins. Skin sensitization test in rats showed that application of the CCK leaching solution caused no obvious skin reddening, regional edema, or skin necrosis. Intracutaneous injection of the leaching solution in rabbits did not induce obvious skin stimulation manifested by intradermal erythema or edema. In acute systemic toxic test, administration of the leaching solution in mice caused no death, organ dysfunction, cyanosis, tremor, severe peritoneal irritation, ptosis, or dyspnoea. In vitro cytotoxicity test indicated that the cell toxicity of the CCK leaching solution was approximately at 0 level.</p><p><b>CONCLUSION</b>CCK contained in the treated chicken calamus easily undergoes hydrolysis to release mainly some peptides which do not induce obvious toxic effects, suggesting the safe potential applications of CCK conduit as a tissue-engineering biomaterial.</p>

Effects of serum of rats with ventilator-induced lung injury on endothelial cell permeability and its mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of inflammatory mediators in the serum of rats with ventilator-induced lung injury (VILI) on endothelial cellular cytoskeleton and monolayer cellular permeability and explore the molecular mechanism of VILI-induced lung edema.</p><p><b>METHODS</b>Thirty healthy male SD rats were divided into 3 groups, namely group A with normal tidal volume ventilation, group B with high tidal volume ventilation and group C with high tidal volume ventilation plus ulinastatin treatment. The serum was collected from each rat after ventilation and added into endothelial cell line ECV-304 culture medium, and 2 h later the changes of F-actin and cell permeability were observed.</p><p><b>RESULTS</b>Compared to sera from rats with normal tidal volume ventilation, the sera of rats with high tidal volume ventilation caused obvious reorganization of actin cytoskeleton with weakened fluorescent intensity at the peripheral filament bands and formation of long and thick stress fibers in the center, which resulted in endothelial contraction and increased cell permeability. Pretreatment with ulinastatin could lessen these changes significantly. The percentage in change of permeability coefficient (Ppa%) after stimulation with the sera of rats in groups A, B and C was (6.95+/-1.66)%, (27.50+/-7.77)%, and (17.71+/-4.66)%, respectively, showing statistically significant differences (P<0.05).</p><p><b>CONCLUSION</b>The pro-inflammatory mediators in the serum of rats with high tidal volume ventilation increases endothelial cell permeability by reorganizing actin cytoskeleton, and pretreatment with ulinastatin can lessen the hyperpermeability by inhibiting multiple pro-inflammatory mediators.</p>

Collagen gel coating or cyclosporine A for improving histocompatibility of chicken calamus keratin / 南方医科大学学报

<p><b>OBJECTIVE</b>To improve the histocompatibility of chicken calamus keratin (CCK) graft by collagen-gel coating or using of cyclosporine A (CsA).</p><p><b>METHODS</b>Thirty SD rats were equally randomized into 5 groups, and in 4 of them, CCK implantation into the bilateral erector spinae was performed on different treatment protocols. In group A, the rats received daily intraperitoneal injection of CsA (5 mg/kg) for two consecutive weeks after CCK implantation; in group B, CCK was soaked in CsA (2.5 mg/ml) solution at 4 degrees Celsius; for 48 h before grafting; in group C, CCK coated with collagen gel was grafted; and in group D, only CCK was implanted. Rats in the fifth group received only cutaneous incision as well as muscular dissection to serve as the blank control. CCK degradation and its effect on the surrounding tissues were observed at 2, 4 and 8 weeks after grafting. Immunohistochemistry was performed to identify T lymphocyte infiltration in the host tissues.</p><p><b>RESULTS</b>All the rats survived the operation. Numerous macrophages, especially multinucleated giant cells occurred on the peripheral of the CCK grafts, and small degraded CCK pieces were observed in their cytoplasm. Only a few inflammatory cells were seen in the host tissues. At 2, 4 and 8 weeks after CCK implantation, only a few CD3-positive cells were found in all the groups, and in group A and B, the density of T lymphocytes was significantly lower than that in group D, and there was no significant difference between group A and the blank control group.</p><p><b>CONCLUSIONS</b>CsA significantly improves the histocompatibility of CCK material, and short-term systemic CsA administration achieves the best results. Macrophages, especially multinucleated giant cells participate in CCK degradation in vivo.</p>

Preparation and bioactivity of human hair keratin-collagen sponge, a new type of dermal analogue / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a three-dimensional porous film of human hair keratin (HHK)-collagen sponge complex for use as a dermal substitute.</p><p><b>METHODS</b>The three components F, B, and Z derived from healthy human hair were weaved into a meshwork and integrated with purified soluble type I collagen extracted from bovine tendons to prepare a highly porous film with vacuum freeze-drying followed by secondary cross-linking with glutaraldehyde. The film was grafted beneath the dorsal skin in 21 SD rats (experimental group), with simple collagen sponge serving as the negative control. The rats receiving surgical operation but without graft served as the blank control. The graft and its surrounding tissue were harvested on days 3, 7 and at weeks 2, 4, 6, 8, 12 after implantation for evaluation of tissue compatibility, vascularization and degradation.</p><p><b>RESULTS</b>The prepared collagen sponge film was semitransparent and porous. Three to 7 days after grafting, inflammatory reaction was relieved gradually, and several fibroblasts and blood vessels were found adherent to the grafts in the experimental groups. At week 4, the wounds healed in the experimental groups, and the fibroblasts were actively secreting collagen and the film degraded obviously with the appearance of elastic fibers. At weeks 6 and 8, new collagen fibers thickened and assumed regular arrangement, and the collagen sponge films disappeared completely. In the control groups, the changes were less obvious and total HHK degradation occurred till week 12.</p><p><b>CONCLUSION</b>The degradable and absorbable HHK-collagen sponge film has relatively satisfactory tissue compatibility and can accelerate wound healing by stimulating cell proliferation and vascularization, showing the potential as an optimal dermal substitute.</p>

Effects of serum of the rats ventilated with high tidal volume on endothelial cell permeability and therapeutic effects of ulinastatin / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>With the widespread use of ventilators in treating critically ill patients, the morbidity of ventilator-induced lung injury (VILI) is increasing accordingly. VILI is characterized by a considerable increase in microvascular leakiness and activation of inflammatory processes. In this study we investigated the effects of inflammatory mediators in VILI rat serum on endothelial cytoskeleton and monolayer cellular permeability, as well as the therapeutic effect of ulinastatin, to explore the pathogenesis and the relationship between biotrauma and lung oedema induced by VILI.</p><p><b>METHODS</b>Thirty healthy male Sprague-Dawley rats were randomly divided into three groups: group A (normal tidal volume ventilation), group B (high tidal volume ventilation) and group C (high tidal volume ventilation plus ulinastatin). The serum of each rat after ventilation was added to endothelial cell line ECV-304 medium for two hours to observe the effects of serum and/or ulinastatin on endothelial fibrous actin and permeability.</p><p><b>RESULTS</b>Compared to rats ventilated with normal tidal volume, serum of rats ventilated with high tidal volume caused a striking reorganization of actin cytoskeleton with a weakening of fluorescent intensity at the peripheral filament bands and formation of the long and thick stress fibres in the centre resulting in endothelial contraction and higher permeability. Prior treatment with ulinastatin lessened the above changes significantly. The changes of permeability coefficient of endothelial permeability after group A, B or C rats serum stimulation were (6.95 +/- 1.66)%, (27.50 +/- 7.77)% and (17.71 +/- 4.66)% respectively with statistically significant differences (P < 0.05) among the three groups.</p><p><b>CONCLUSIONS</b>The proinflammatory mediators in the serum of the rats given high tidal volume ventilation increases endothelial permeability by reorganizing actin cytoskeleton, and pretreatment with ulinastatin lessens the permeability by inhibiting of proinflammatory mediators.</p>

Ooplasmic transfer: problems and prospects / 南方医科大学学报

Cytoplasmic transfer between human oocytes, which represents a complete cytoplasmic exchange, has been performed recently as a means to improve the outcome of assisted reproduction and becomes a hotspot of researches. Many studies have indicated that mitochondria in the oocytoplasm obviously affect fertilization of the oocytes and early embryo development. However, ooplasmic transfer can lead to mitochondrial DNA heteroplasmy and the prospect of mitochondrial heteroplasmy and its potential problems necessitate further studies. The authors reviews the ooplasmic transfer, the relation between ooplasm and fertilization and embryo development, and the mitochondrial heteroplasmy. The authors also propose a new theory of "reverse cloning technique".