Effect of methyl eugenol on nasal mucosal aquaporin 5 in rats with allergic rhinitis / 北京大学学报(医学版)

OBJECTIVE@#To observe the effect of methyl eugenol on the expression of aquaporin (AQP) 5 in nasal mucosa of rats with allergic rhinitis and to explore its significance.@*METHODS@#In the study, 128 Wistar rats were randomly divided into normal control group, AR model control group, budesonide positive control group, 80 mg/kg group, 40 mg/kg group, 20 mg/kg group and 10 mg/kg group, and ovalbumin (OVA) was used to establish the model of allergic rhinitis. After successful modeling, castor oil, budesonide and corresponding doses of methyl eugenol were given respectively. After 1, 2 and 4 weeks of administration, the distribution of AQP5 in nasal mucosa was observed by immunohistochemistry. The expression of AQP5 in nasal mucosa of each group was compared by Western blotting. The expression of AQP5 mRNA was compared with real-time PCR.@*RESULTS@#AQP5 was mainly located in the glandular epithelium and ductal epithelial cell membrane and cytoplasm. The expression of AQP5 and AQP5 mRNA in nasal mucosa of the rats in the model control group was lower than that in the normal control group (P<0.05). AQP5 and AQP5 mRNA in nasal mucosa of the rats in each treatment group were higher than those in the model control group in varying degrees. The expression of AQP5 in the budesonide group was not significantly different from that in the normal control group 1, 2 and 4 weeks after drug intervention (P>0.05), but there was significant difference between the budesonide group and the model control group (P<0.05). The expression of AQP5 mRNA in the budesonide group was significantly different from that in the normal control group and the model control group (P<0.05).After 2 weeks of intervention, the expression of AQP5 in each dose group of methyleugenol was not significantly different from that in the budesonide group (P>0.05). After 1 week of intervention, there was no significant difference in AQP5 mRNA between the 20 mg/kg group and the normal control group (P>0.05), but there was significant difference between the 20 mg/kg group and the model control group (P<0.05).@*CONCLUSION@#Methyl eugenol may increase the degree of edema of the nasal mucosa by reducing the expression of AQP5 and reduce the secretion of glands, thus alleviating the symptoms of allergic rhinitis, sneezing and runny nose.

Effects of Col-Tgel, a bioactive material, on human tissue stem cells / 中国组织工程研究

BACKGROUND:The traditional two-dimensional culture system has been widely used in the in vitro culture of human tissue stem cells,but it cannot really simulate the three-dimensional physiological microenvironment in the body, which is not conducive to the study of the biological behavior of human stem cells. OBJECTIVE: To detect the effect of the bioactivity of Col-Tgel in human hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs)in vitro and in vivo,by constructing a three-dimensional culture system stimulating the physiological microenvironment of the body. METHODS:(1)In vitro co-culture:Green fluorescent protein labeled MSCs(MSCs-GFP)and human umbilical cord blood CD34+cells were co-cultured in Col-Tgel for 3 days (three-dimensional culture group). Human umbilical cord blood CD34+cells were cultured in Col-Tgel for 3 days as single culture group. MSCs-GFP and human umbilical cord blood CD34+cells were co-cultured in Transwell chamber for 3 days as two-dimensional culture group. Human umbilical cord blood CD34+cells were cultured routinely as control group. The percentage of CD34+CD38-CD45RA-CD90+cells in each group was measured by flow cytometry. In situ immunofluorescence staining was used to detect the activity of cells that were co-cultured in Col-Tgel.(2)In vivo transplantation:NOD/SCID mice subjected to 24-hour X-ray irradiation were divided into two groups: in experimental group, MSC-GFP cells were resuspended in Col-Tgel and transplanted into the tibia of NOD/SCID mice; in control group, MSCs-GFP were resuspended in PBS and transplanted into the tibia of NOD/SCID mice. The MSC-GFP growth in the bone marrow was detected by two-photon/confocal microscopy at 3 days post transplantation. RESULTS AND CONCLUSION: (1) After co-culture in Col-Tgel for 3 days, the percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was 2.8 times that of the two-dimensional culture group, indicating that the MSCs significantly promoted the expansion of CD34+CD38-CD45RA-CD90+cells in the Col-Tgel. The percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was increased by 4.5 times compared with the single culture group and increased by 1.5 times compared with the control group. Immunofluorescence staining showed that the cell viability of human MSCs and human umbilical cord blood CD34+cells was not affected after co-cultured in Col-Tgel for 3 days.In the in vivo transplantation experiment,MSC-GFP cells could survive in the medullary cavity.In summary, Col-Tgel provides a new strategy for stem cell culture and in vivo growth by forming a three-dimensional system similar to the physiological environment in vivo.

Establishment of Integration-Free Human Induced Pluripotent Stem Cells from A Patient with Primary Myelofibrosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the primary myelofibrosis (PMF)-induced pluripotent stem cell line (iPSC) by means of iPSC techinique so as to provide a experimental model for studying the blood disease mechanisms.</p><p><b>METHODS</b>Induced pluripotent stem cells were generated from mononuclear cells isolated from a PMF patient with JAK2(V617F) mutation by using episomal vectors.</p><p><b>RESULTS</b>PMF-derived iPSC was established from the patient with JAK2(V617F) gene mutation. The PMF-iPSC could be stably passaged, highly expressed pluripotent genes as human embryonic stem (ES) cells, and were able to form teratoma in NOD/SCID mice in vivo. H & E staining of the teratoma showed the presence of tissue type derived from all three embryonic germ layers. Sanger sequencing confirmed that PMF-derived iPSC carried different allele burdens of JAK2(V617F) gene mutation.</p><p><b>CONCLUSION</b>The interation-free iPSC from primary myelofibrosis patient in vitro has been established. This PMF-derived iPSC line provides a valuable tool for studying the pathogenesis, screening of chimical drugs and realizing the standard therapy of PMF.</p>

Effect of 18-β Glycyrrhetinic Acid on the Endoplasmic Reticulum of Nasal Epithelial Cells in Allergic Rhinitis Model Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of 18-β glycyrrhetinic acid (GA) on the endoplasmic reticulum of nasal epithelial cells in allergic rhinitis (AR) model rats.</p><p><b>METHODS</b>Totally 96 Wistar rats were randomly divided into the blank group, the AR model group, the loratadine group, the GA group, 24 in each group. AR models were established by peritoneally injecting ovalbumin (OVA). Morphological scoring was performed. GA at 21. 6 mg/kg was intragastrically administered to rats in the GA group. Nasal mucosal tissues were taken for electron microscopic examinations at the second, fourth, sixth, and tenth week after drug intervention.</p><p><b>RESULTS</b>The overlapping score was 2.10 ± 0.45 in the blank group, 5.10 ± 0.56 in the loratadine group, 5.10 ± 0.56 in the AR model group, 5.20 ± 0.78 in the GA group, showing statistical difference when compared with the blank group (P < 0.01). Results under transmission electron microscope showed that the number of the endoplasmic reticulum increased in the AR model group, with obvious cystic dilatation, a lot of vacuole formation, and degranulation. A large number of free ribosomes could be seen in cytoplasm. With persistent allergen exposure, changes mentioned above was progressively aggravated in the endoplasmic reticulum of nasal mucosal epithelium in the AR model group. But the dilation of endoplasmic reticulum, vacuole formation, and degranulation were relieved in the GA group, and got close to those of the blank group.</p><p><b>CONCLUSION</b>18-β GA could improve the expansion, vacuolization, and degranulation of the endoplasmic reticulum of nasal epithelial cells in AR model rats.</p>

Effects of catalase on the engraftment of human hematopoietic stem cells in NOD/SCID mice / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of catalase (CAT) on engraftment of human hematopoietic stem cells (HSC) by co-transplanting umbilical cord-derived mesenchymal stem cells (UC-MSC) with over-expressed CAT and human HSC into NOD/SCID mice.</p><p><b>METHODS</b>The UC-MSC cultured in vitro were transfected by the retrovirus containing green fluorescent protein (GFP) and GFP-CAT genes respectively. MSC-GFP and MSC-GFP-CAT cell lines were sorted by flow cytometry. Co-culture and co-transplant experiments were performed to detect the effects of CAT on expansion and engraftment of human HSC.</p><p><b>RESULTS</b>The percentage of GFP(+) cells were approximately 97.6% and 96.8% after sorting. The mRNA expression of CAT in MSC-GFP-CAT was 23.9-fold higher than that in UC-MSC. The activity of CAT in UC-MSC, MSC-GFP, MSC-GFP-CAT cells were 19.5, 20.3 and 74.1 Unit respectively. There was no significant differences in the percentage of CD34(+) cells between 3 groups in co-culture experiment. And the percentage of human CD45(+) cells in NOD/SCID mice were (3.22 ± 3.1)%, (4.26 ± 3.56)% and (7.37 ± 4.51)% respectively.</p><p><b>CONCLUSION</b>MSC-GFP-CAT significantly improves the engraftment of human HSC in NOD/SCID mice, whereas co-culture with the MSC-GFP-CAT can not promote the expansion of HSC in vitro.</p>

An improved method for generating integration-free human induced pluripotent stem cells / 中国实验血液学杂志

The genome instability and tumorigenicity of induced pluripotent stem cells (iPSC) hinder their great potentials for clinical application. Using episomal vectors to generate iPSC is the best way to solve safety issues at present. This method is simple and the exogenous gene was not integrated into the host genome. However, the reprogramming efficiency for this method is very low and thus limits its usage. This study was purposed to improve episomal method for generating induced pluripotent stem cells from cord blood mononuclear cells (CB MNC), to establish integration-free iPSC technology system, and to lay the foundation for individualized iPSC for future clinical uses. To improve the reprogramming efficiency for iPSC, episomal method was used at various combinations of episomal vectors, pre-stimulating culture mediums and oxygen condition were tested to optimize the method. The results showed that using erythroid culture medium for culturing 8 days, transfecting with episomal vectors with SFFV (spleen focus forming virus) promoter under the hypoxic condition (3%), CB MNC could be mostly efficiently reprogrammed with the efficiency 0.12%. Furthermore, the results showed that erythroblasts (CD36(+)CD71(+)CD235a(low)) were the cells that are reprogrammed with high efficiency after culture for 8 days. It is concluded that a highly efficient and safe method for generation of integration-free iPSC is successfully established, which is useable in clinical study.

Protein O-GlcNAcylation and cancer / 肿瘤

O-GlcNAcylation is the addition of a single N-acetylglucosamine (GlcNAc) moiety to the hydroxyl groups of serine or threonine residues of nuclear and cytoplasmic proteins. O-GlcNAc plays an important role in the regulation of many biological processes including, but not limited to, cell cycle progression, transcription, translation, signal transduction, stress response, etc. O-GlcNAc plays significant roles in the progression and etiology of various diseases including diabetes, cardiovascular disease, cancer and Alzheimer's disease. The aberrant O-GlcNAcylation in tumor tissues is closely associated with cell proliferation and metastasis. This review is mainly involved the characteristics of protein O-GlcNAcylation alterations in different types of human cancer and the functions of O-GlcNAcylation in oncogenes and tumor suppressor genes. Copyright © 2013 by TUMOR.

Effect of whisker trimming on behavior and barrel cortex of rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the change of behavior, as well as the plasticity of somatosensory cortex after whisker trimming.</p><p><b>METHODS</b>SD rats were divided into 4 groups. Group A is the normal control group; group B: bilateral vibrissotomy on the second postnatal day; group C: unilateral right vibrissotomy on the second postnatal day; group D: right unilateral whisker trimmed during 1-5 days after birth, and leave untreated after the 5th postnatal day. Their body weight, length of the left D2 whiskers was measured on the 30th postnatal day. At the same time, the changes of their behavior (including the slit-detection test, the home exploring behavior and thigmotaxis test) were also recorded on the 30th postnatal day. Cytochrome oxydase histochemistry (CO reaction)was applied to study the development and arrangement of barrel cortex.</p><p><b>RESULTS</b>In the slit-detection test, control rats could find and get into the right slit very quickly. The rats in group B could get into the slit only if their noses touched the slit. The rats in group C couldn't identify the slit by right face, but if they turned their body and touched the slit with the left whiskers, they could get into the slit very quickly. The behavior of rats in group D was similar to that in group C. The time spent for finding out the right slit of the rats in group A, B, C was obviously longer than that of group A (P < 0.01, P < 0.05, P < 0.01). In the exploring behavior and thigmotaxis test, the time for left thigmotaxis, right thigmotaxis and total thigmotaxis of rats in group B was longer than that of control animals. The time for right thigmotaxis of group C was significantly shorter than that of group A (P < 0.05). Both the weight of the rats and the length of left D2 whiskers of rats in all the four groups had no significant difference. CO reaction showed that the barrels became smaller, the septum was not clear, the arrangement of the barrels was not tidy in the mice whose right whiskers were trimmed from 2-30 days after birth.</p><p><b>CONCLUSION</b>Deafferentation doesn't change the body weight and length of the whiskers left. But the stimulation of whiskers is important for rodent especially in thigmotaxis and exploring behavior. Deafferentation can also induce the plastic change of barrel cortex.</p>

A clinical study of fungal infection in burn patients / 中华烧伤杂志

<p><b>OBJECTIVE</b>To address the features of the fungal infection after burn injury in clinic.</p><p><b>METHODS</b>Three thousand nine hundred and nine burn patients admitted to our institute from Jan. 2003 to Dec. 2006 were involved in this study. Two thousand two hundred and seventy-one samples were harvested for fungal detection by culture from 467 patients suspected to be infected by fungi based on their clinic manifestations. The collected samples included wound tissue, blood, urine, stool, sputum, catheters and others. The antibiotic sensitivity of the identified fungi were determined by routine method. When same kind of fungus was found from different samples taken from one patient, it was recorded as one positive sample. The samples were ranked in an ascending order as wound secretion, stool, urine, sputum and bronchial alveolar lavage fluid, arteriovenous catheter or urinary catheter, blood. Only the positive sample of the highest rank source was recorded as the positive strain of fungus from this particular patient.</p><p><b>RESULTS</b>It was found 61 fungal positive samples from the 2271 samples collected. Out of 467 patients, 38 strains of fungi were detected from 36 burn patients during the investigated period, the incidence was 0.92% (36/3909). The most three commonest types among the identified 38 strains of fungi were Candida tropicalis (42.1%), Candida albicans (31.6%) and Candida famata (T. Famata, 10.5%). The drug sensitivity tests demonstrated that most of the strains detected in this investigation, with the exception of candida glabrata, were sensitive to most of the routine antimycotics agents such as Amphotericin B, Fluconazole, and Itraconazole etc. Among the 36 fungus positive patients, in 18 patients the burn area exceeded 80% TBSA, 12 patients with 50%-79% TBSA, 4 patients with 30%-49% TBSA, and in 2 patients the burn area was smaller than 30% TBSA. It was found most of the fungal infections (77.78%) occurred 2 weeks after burn injury, and 8 of the 36 fungus-infected patients died (the mortality was 22.22%). Conclusions Further examinations are necessary to confirm the diagnosis in burn patients suspected to have fungal infection. Once fungal infections are confirmed, antimycotic therapy must be started immediately.</p>

Study on the situation of plague in Junggar Basin of China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the distribution, fauna, population structure of host animals and their parasitic fleas as well as popular dynamic of animal plague of natural plague foci in Junggar Basin.</p><p><b>METHODS</b>Sample materials and data of animals and vector insects were collected using ecological methods and the population structures were analyzed statistically. F1 antibody of Yersinia pestis in rodents' serum and organ suspension was detected by means of IHA while the pathogen of Y. pestis in rodents and vector insects was detected by means of aetiological detections and the isolated Y. pestis was detected using biochemical methods.</p><p><b>RESULTS</b>The small mammals which were found in Junggar Basin belonged to 17 species of 11 genera 7 families. Of them, 13 species of rodents were included whose parasitic fleas belonged to 19 species of 10 genera 8 families. The average coverage of Rhombomys opimus hole-community was 22.5% in Junggar Basin with the average density of R. opimus hole-community was 15.9/hm2 and the average rate of habitat of the hole-community was 70.2%. In the R. opimus community, the average density of rodents was 3.1/hole-community, and 34.4/hm2 in the nature plague foci. In the population structure of the hole-community of R. opimus, R. opimus accounted for 72.9% in the total captured rodents, Meriones meridianus was 24.5% while the others were 2.6%. In the nocturnal community of rodents, M. meridianus accounted for 64.0% in total captured rodents, Dipus sagitta was 15.1%, M. erythrourns was 7.5% and the others were 13.4%. In the rodents community of Junggar Basin, the rate of R. opimus with fleas was 84.9%, which was the highest, followed by M. tamariscinus, Euchoreutes naso and M. erythrourns, with the rates as 71.4%, 66.7% and 62.7% respectively. The rate of M. meridianus with fleas was 38.3%. There were 16 species of parasitic fleas in R. opimus, with the total flea index as 8.58 and the dominant species was Xenopsylla skrjabini. There were 17 and 16 kinds of fleas in M. erythrourns and M. meridianus respectively with the total flea index were 1.59 and 1.15, with dominant fleas were Nosopsyllus laeviceps and X. skrjabini. The serum and organ suspension of 3179 rodents which belonged to 12 species were detected by means of IHA, of them 174 samples were positive and the positive rate was 5.5%. There were 1356 samples of R. opimus in these materials, and 164 were positive, accounted for 12.1%. The samples of M. meridianus were 1255, with 9 positive, accounted for 0.7%. The samples of D. sagitta were 116 with 1 positive and the rate was 0.9%. The samples of other rodents were 452 but were all negative. There were in total 2975 organs collected from rodents, when detected by methods of isolated of Y. pestis. 15 strains of Y. pestis were isolated from 1243 R. opimus, and 2 strains isolated from 1230 M. meridianus. A total number of 11 647 fleas from rodents were detected by methods of isolated of Y. pestis in which 1 strain of Y. pestis was isolated from 4713 X. skrjabini, and 6 were isolated from 2101 Xenopsylla minax, 1 from 328 Xenopsylla conformis conformis and 1 from 250 Echidnophaga oschanini. Among the other 4255 fleas, none was isolated. The biochemical properties of these Y. pestis which isolated from Junggar Basin were positive of Maltose, Ejiao sugar and Glycerol, and negative of Rhamnose and Nitrogen, which were all strongly poisonous to mouse.</p><p><b>CONCLUSION</b>The natural plague foci in Junggar Basin spread all over the whole Junggar Basin. There were animal plague cases found in 12 counties (cites) while Karamy, Bole, Jimusaer and Qitai were confirmed as plague foci counties (cities). Animals and vector insects of the foci were complicated but the ecological system was stable. R. opimus was recognized as the dominant host animal and its biochemical type belonged to the Middle Ages, suggesting that the foci was a new type of natural plague foci.</p>

Expression of neutral endopeptidase and motility-related protein-1 in cutaneous malignant melanoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of neutral endopeptidase (CD10) and motility-related protein-1 (CD9) in malignant melanoma and their clinical significance.</p><p><b>METHODS</b>Immunohistochemical study for CD10 and CD9 using Streptavidin-biotin complex technique was carried out in 48 cases of primary cutaneous malignant melanoma (CMM), 23 cases of metastatic melanoma and 23 cases of benign nevus.</p><p><b>RESULTS</b>The positivity rate of CD10 was highest in metastatic melanoma and lowest in benign nevus (P < 0.01). In contrast, the positivity rate of CD9 in metastatic melanoma was lower than that in CMM (P < 0.05). The expression of CD9 was inversely correlated with that of CD10 in malignant melanoma (CMM: r = -0.40, P = 0.005; metastatic MM: r = -0.44, P = 0.034). The expression of CD10 and CD9 in CMM also correlated with tumor histology, Clark's level of invasion and presence of nodal metastasis. A similar relationship was also observed for CD10 and CD9 expression in stromal fibroblasts of CMM (r = -0.43, P = 0.007).</p><p><b>CONCLUSIONS</b>CD10 and CD9 expression correlates with the invasiveness and metastatic potential of malignant melanoma; both factors may demonstrate a counteracting effect. These two markers have potential implications in prognostic assessment of CMM. Stromal fibroblasts may also play an important role in the progression of CMM.</p>

Effects of resveratrol on inflammatory process induced by focal cerebral ischemia-reperfusion in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effects of resveratrol on inflammatory process induced by focal cerebral ischemia-reperfusion in rats.</p><p><b>METHOD</b>Rats were pretreated with resvreratrol at the dose of 10, 20, 40 mg kg(-1) for 7 days and then subjected to cerebral ischemia/reperfusion induced by a middle cerebral artery occlusion (MCAO). The infarct volume and the neurological deficit were determined by the method of TTC (2, 3, 5-triphenylterazolium chloride) staining and Longa's score. The permeability of blood-brain barrier (BBB) was evaluated by measurement of the evans blue (EB) content in the brain with spectrophotometer. The content of interleukin-lbeta, interleukin-6 (IL-6, IL-1beta) in serum and tumor necrosis factor-alpha (TNF-alpha), myeloperoxidase (MPO) in brain were determined by radio-immunoassay and ELISA assay.</p><p><b>RESULT</b>Resveratrol reduced infarct volume, ameliorated the neurological deficit and the permeability of BBB, the content of IL-6, IL-1beta in serum and TNF-alpha, MPO activity in brain tissue also were significantly decreased.</p><p><b>CONCLUSION</b>These results showed that resveratrol had protective effects on cerebral injury by inhibiting the releasing of the inflammatory mediators after ischemia/reperfusion injury.</p>

A follow-up study of family intervention for rehabilitating chronic outpatients with schizophrenia in the country / 中国康复理论与实践

@#ObjectiveThe trial was to examining the effectiveness and cost effects of family intervention for rehabilitating chronic outpatients with schizophrenia in the country.Methods90 subjects were randomly assigned to the family intervention group and the control group. Both groups received the same treatments, but the family intervention courses mainly containing mental health education were given to the family intervention group for one year. During the time, all subjects were evaluated with standard rating scales and self made criteria. ResultsThe family intervention group demonstrated clinical results significantly superior to those of the control group on overall improvement according to the scores on the SDSS, the SAPS, the SANS and the MRSS. Substantially, the direct and indirect average cost in the family intervention group was significantly lower than those of the control group. Conclusions Family intervention is effective not only in making the schizophrenics recover from illness but also in both increasing their social functions and reducing their medical cost.