[Objective]To investigate the effect of coagulation of intervertebral disc nucleus pulposus(IDNP) by alum.[Method]In vitro,20 canine intervertebral discs(I D) were divided into 4 groups,and immersed into 2.5%,5%,10% alum solution and normal saline respectively.Another 16 I D also randomized into 4 groups and were injected into I D with corresponding solution.In vivo,6 canine ID were grouped into 4:A.blank control;B.one injection of alum;C.two points injection of alum;D.injection of N.S.Experimental dogs were sacrificed and disc samples were taken at 3 days,2 wks,and 1 month post-injection.Gross observation,histological exam by microscopy and scan electron microscopy(SEM) and histochemical test were performed.[Result]NS did not coagulate the IDNP in vivo and in vitro,while in vivo,coagulation was seen around the injection points,two points with two coagulation blocks,and grew with time culminating on one month after injection,but there was no coagulation appeared after alum injection.In vitro,histological study showed aboundant collagenous fibers gathered in coagulation lock representing manifold function of mesenchyme.[Conclusion]The alum can make the IDNP to be coagulated in vivo,the mechanism of coagulation is the collagenous fiber proliferation and fibrosis due to manifold function of mesenchyme.

BACKGROUND: The main pathological change of intervertebral disc herniation is that nucleus pulposus protrudes from ruptured annulus fibrosus, thus we can hypothesize that it is possible to prevent disc herniation prior to its protrusion by coagulating it as a whole.OBJECTIVE: To observe the coagulation effects of intervertebral disc nucleus pulposus by alum solution.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed between September 2002 and April 2003 at Department of Animal Experiment, Peking University First Hospital, Beijing China.MATERIALS: Twenty-six healthy adult hybrid dogs, 9 for in vitro experiment and 17 for in vivo experiment, weighing 16-21kg, with no restrictions on male and female, were obtained from Department of Animal Experiment, Peking University First Hospital, Beijing, China.METHODS: Twenty canine in vitro intervertebral discs obtained from 5 dogs were randomly divided into 4 groups, with 5 discs in each group, and were put into 2.5%, 5%, 10% alum solutions and 0.9% physiological saline, respectively. Effects of disc coagulation were observed after immersing for 1 day and 10 days, respectively. Another 16 in vitro intervertebral discs obtained from 4 dogs, composed of L2/3, L3/4, L4/5, L5/6, were also injected with the above 4 experimental solutions, 0.15ml, respectively. Sixty-eight in vivo intervertebral discs were obtained from 17 dogs and divided into 4 groups: blank control group, physiological saline group, 10% alum solution+one puncture point group, and 10% alum solution+two puncture points group, 17 discs in each group. Harvesting time: 3 days, 2 weeks, 1 month and 3 months postoperation.MAIN OUTCOME MEASURES: Effects of alum solutions on the coagulation of the intervertebral discs and related histological changes were observed and an alum solution of suitable concentration was preliminarily selected. General observation, light microscopic observation and scanning electron microscopic observation were made of the nucleus pulposuses.RESULTS: In the in vitro and in vivo experiments, it was found that physiological saline did not produce the effect of coagulation on the nucleus pulposus, while immersion in the alum solution induced nucleus pulposus coagulation in the in vitro intervertebral discs. Also, as the concentration of the alum solution increased, the volume of the coagulated nucleus pulposus gradually decreased. After alum solution was injected into the in vitro intervertebral discs, no nucleus pulposus coagulation appeared. When the 10% alum solution was injected into the in vivo intervertebral discs, nucleus pulposus coagulation occurred, with the strongest coagulation effect reached at 1 month postoperation. This was manifested in the agglutination reaction centered around the puncture point. When there were 2 puncture points, 2 coagulated lumps might appear. There was an increase in the mesenchymal component of the coagulated nucleus pulposus. Histochemical and scanning electron microscopic examinations confirmed the proliferation of large numbers of collagen fibers in the mesenchyme.CONCLUSION: Alum can promote nucleus pulposus to coagulate around the injection point. This may be related to the increase of collagens and the fibrosis resulting from stimulation of the nucleus pulposus by alum solution.

Over 3 000 pictures on five major parasites(schistosome,filaria,hookworm,leishmania and plas-modium)collected between 1950 and 1990 were edited and a searching system was established. The data can be used with a network-based version through a LAN system in the Institute. The adoption of Digital Computerized Management makes it possible for sharing resources in human parasitology.

Objective To discover the role of MCPH1 in DNA double-strand damage induced by ionizing radiation and its relationship with H2AX in esophageal cancer cell ECA109. Methods ECA109 cancer cells accepted 8 Gy 1 h after irradiation were collected for protein extraction and immunofluorescence then MCPH1 and H2AX protein expression and nuclear foci changes were observed. A stable low expression of H2AX cell lines was established and MCPH1 and H2AX protein expression and nuclear foci changes induced by ionizing radiation after silence H2AX were detected. Results (1)A stable low expression of H2AX cell lines in ECA109 cells was successfully constructed. (2)Ionizing radiation could cause the increase of r-H2AX and MCPH1 protein expression, as the same as nuclear focus increase of r-H2AX and MCPH1. (3)The protein level and nucleus focus of r-H2AX and MCPH1 were significantly reduced in ECA109 after silence H2AX. Conclusion MCPH1 is the part of DNA damage response triggered by ionizing radiation and is located in damage response downstream and can be regulated by H2AX.

Objective To evaluate the bone repair capacity ofpolylactic acid-polyglycolic acid copolymer (PLGA)/collagen type Ⅰ (CoI) microspheres combined with BMSCs after being injected in intertrochanteric bone defect of osteoporotic female rats.Methods Prepared PLGA microspheres.The microspheres were coated with Col.BMSCs of the third passage were cultured with PLGA/CoI microspheres.Forty 3-month-old female SD rats were ovariectomized to establish osteoporotic animal models.The osteoporotic rats were randomly divided into 5 groups,including SHAM group,PBS group,Cell group,MS group and Cell+ MS group.There were 8 rats in each group.Different material was injected into the intertrochanteric bone defect site which was made with electric drill.Four rats of each group were sacrificed at 1 month and 3 months post-operation.The fenora were taken to measure the intertrochanteric bone mineral density (BMD) with DEXA and evaluate trabecular stucture with Micro CT.Results After 7 days of coculture,BMSCs seeded on PLGA/CoI microspheres had nice adherance and proliferation.There was no difference of BMC and BMD among all groups at 1 month post-operation.Tb.Th of Cell+MS group was higher than that of PBS group and MS group at 1 month post-operation.％Tb.Ar of Cell+MS group was higher than that of Cell group and MS group at 1 month post-operation.Tb.Sp of Cell+MS group had a tendence to decrease compared with other groups but there was no statistical difference at 1 month post-operation.After 3 months of operation,the BMC of Cell+MS group had a tendence to increase compared with that of PBS group and MS group but showed no statistical difference.BMD and Tb.Th of Cell+MS group was higher than those of other groups.％Tb.Ar of Cell+MS group was higher than that of SHAM group and PBS group.Tb.Sp of Cell+MS group had a tendence to reduce compared with other groups but showed no statistical difference.Conclusion The bone defect of osteoporotic site can be repaired 1 month after the injection of the PLGA/CoI microspheres combined with BMSCs.The trabecular reconstruction and bone quality of osteoporotic site can be improved 3 months after the injection.

Objective To screen the glycoproteins as biomarkers for intracranial aneurysm (ⅠA) in cerebrospinal fluid (CSF) and evaluate the specificity and sensitivity of the biomarker candidates.Methods A complementary proteomic approach integrated with multidimensional chromatography was employed to simultaneously measure relative changes in the gylcoproteins of cerebrospinal fluid (CSF) obtained from patients with ruptured ⅠA (RIA) and unruptured ⅠA (UIA) compared to the healthy controls (HC) and disease controls (DC).One protein-receptor tyrosine kinase Axl with a unique change in RIA was validated in CSF and plasma.The sensitivity at 95％ specificity of Axl in CSF and plasma was evaluated with receiver operating characteristic curve (ROC curve).Results Firstly,a total of 294 glycoproteins were identified in human CSF with believable evidence.Secondly,the proteomic findings showed the quantitative changes in RIA and UIA as compared to HC and DC.Of 294 identified CSF proteins,59,24 and 33 proteins displayed quantitative changes unique to RIA,UIA or IA,respectively.At last,one of these unique proteins-receptor tyrosine kinase Axl with unique increase in RIA was confirmed both in CSF and plasma.ROC curve analysis showed that the sensitivity at 95％ specificity of Axl in CSF to differentiate RIA from UIA was 60％.When compared to CSF,the sensitivity at above setting in plasma to differentiate RIA from HC was 40％ and to differentiate RIA from UIA was 25％.Conclusions A glycoprotein biomarker Axl might be used as a promising biomarker to predict the rupture of ⅠA.The further investigation of the relations between Axl and IA formation as well as rupture might help to elucidate the underlying pathogenesis and find new therapeutic targets.

We report a case of cutaneous and subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei after renal transplantation in Guangdong. A 66-year-old man who had a renal transplantation 6 years ago was admitted in October 2011 for the presence of 16 nodules (0.5-1.5 cm) found on his right middle finger, wrist and forearm for 5 months. Microscopic examination of the purulent exudate showed segmented and branched brown mycelium, and tissue biopsy and PAS staining showed fungal hyphae. The isolate was processed for morphological identification and molecular sequence analysis. A black colony was found after culture of the isolate on SDA at 26 degrees Celsius;, and small culture identified the isolate as Exophiala jeanselmei. ITS sequence analysis of the isolate showed a 100% homology with Exophiala jeanselmei. E-test strip was used in drug sensitivity test, and the isolate was sensitive to amphotericin B, voriconazole, itraconazole and fluconazole, but resistant to 5-flucytosine and caspofungin. Good response was obtained with surgical intervention, local injection and systemic antifungal treatment.
Aged ; Exophiala ; pathogenicity ; Humans ; Kidney Transplantation ; adverse effects ; Male ; Phaeohyphomycosis ; etiology ; Postoperative Complications

OBJECTIVES: To study the effects of 17β-estradiol (E2) on the regulation of the proliferation of condylar chondrocytes and provide a preliminary discussion on the role of phosphorylate-mammalian target of rapamycin (p-mTOR) in this regulatory process. METHODS: Condylar chondrocytes were isolated from 6-week-old female rats for primary culture. Drug treatment with different concentrations of E2 and/or rapamycin (RAPA) was carried out on second-generation cells. Cell Counting Kit 8 was used to measure the cell viability of condylar chondrocytes after culture for 24, 48, or 72 h, and reverse transcription-polymerase chain reaction (RT-PCR) was applied to detect the relative gene expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), collagen type Ⅱ (COLⅡ), autophagy-related gene 6 (Beclin-1), and autophagy-related gene 5 (ATG-5). Western blot was employed to determine the relative protein expression of ERα, ERβ, Beclin-1, lipid-modified light chain 3B (LC3-Ⅱ), and p-mTOR. RESULTS: E2 could significantly promote the proliferation of chondrocytes cultured CONCLUSIONS At a concentration of 10
Animals ; Autophagy ; Cell Proliferation ; Chondrocytes ; Estradiol ; Estrogen Receptor alpha/metabolism* ; Estrogen Receptor beta ; Female ; Phosphorylation ; Rats

Objective:To investigate the effects of different doses of Didangtang on myocardial inflammatory lesions in diabetic mice. Method:Sixty C57BL/6J mice were randomly divided into normal group （n=10） and model group （n=50）. The diabetic mice in the model group were established by intraperitoneal injection of high-fat diet combined with streptozotocin （STZ）. After model reproducing， the mice were fed with high-fat diet. After 8 weeks， the cardiac function of the mice was detected by using an ultrasound imaging platform. If the cardiac function decreased， the diabetic cardiomyopathy mice were modeled successfully. The nonmodel mice were eliminated， and finally 40 model mice were modeled. The rats in the model group were randomly divided into model group， low， medium and high dose of Didangtang group（1.5，3，6 g·kg^-1） and simvastatin group（0.001 5 g·kg^-1） according to heart function， with 8 rats in each group. The cardiac function of mice was detected by ultrasound imaging platform， fiber bragg grating（FBG）， triglyceride（TG） and total cholesterol（TC） were detected by automatic biochemical analyzer， hematoxylin-eosin （HE） staining was used to observe the pathological changes of myocardium， and the levels of NOD-like receptor3（NLRP3）， thiomdoxin interaction protein（TXNIP）， cysteinyl aspartate specific proteinase-1（Caspase-1） and Interleukin-1β（IL-1β） in myocardial tissue， as well as the content of reactive oxygen species（ROS） were detected by Western blot. Result:Compared with the normal control group， the levels of FBG， TC and TG in the model group significantly increased （P<0.01）； the values of EF and FS significantly decreased （P<0.01）； the expression of ROS significantly increased （P<0.05）， and the expressions of NLRP3， TXNIP， Caspase-1 and IL-1β in the myocardial tissue significantly increased （P<0.05）. Compared with the model group， the levels of FBG， TC and TG in the middle and high dose groups of Didangtang and simvastatin groups significantly decreased （P<0.05）； the EF and FS in each dose group and simvastatin group improved （P<0.05）， and the change in the middle dose group was more obvious （P<0.05）. HE staining showed that Didangtang could improve the pathological changes of myocardial tissue in mice， the ROS expression levels of mice in each dose group of Didangtang and simvastatin group significantly reduced， especially in the middle dose group， the expression levels of NLRP3， TXNIP， Caspase-1 and IL-1β in each dose group significantly decreased， and the effect of middle dose of Didangtang on reducing expressions of NLRP3， TXNIP and Caspase-1 in myocardial tissue was more obvious， the effect of high dose of Didangtang on reducing the expression of IL-1β in myocardial tissue was more obvious. Conclusion:Didangtang can improve myocardial inflammatory lesions in diabetic cardiomyopathy mice by inhibiting the activation of NLRP3 inflammasome.

ObjectiveTo observe the effects of modified Shenqiwan on renal function and fibrosis in diabetic nephropathy mice and explore the underlying mechanism based on the glycogen synthase kinase-3β （GSK-3β）/cyclic adenosine monophosphate （cAMP） response element-binding protein （CREB） signaling pathway. MethodFifty male db/db mice and 10 db/m mice were used in this study. The fifty db/db mice were randomly divided into model group， irbesartan group， and low-， medium-， and high-dose modified Shenqiwan groups. The 10 db/m mice were assigned to the normal group. The mice in the low-， medium-， and high-dose modified Shenqiwan groups were administered with modified Shenqiwan in the dosage form of suspension of Chinese medicinal granules by gavage， those in the irbesartan group were given irbesartan suspension by gavage， and those in the normal and model groups were given distilled water of equal volume by gavage. The intervention lasted for 12 weeks. The blood glucose levels， urine albumin-to-creatinine ratio （UACR）， and the protein expression levels of GSK-3β， CREB， transforming growth factor-β₁ （TGF-β₁）， E-cadherin， Vimentin， fibronectin （FN）， plasminogen activator inhibitor-1 （PAI-1）， and Collagen type Ⅳ （Coll Ⅳ） in the mouse kidneys were recorded before and after treatment. The extent of renal pathological damage was also observed. ResultCompared with the normal group， the model group showed significant increases in blood glucose levels， UACR levels， and the protein expression levels of GSK-3β， TGF-β₁， E-cadherin， Vimentin， FN， PAI-1， and Coll Ⅳ in the kidneys （P<0.05）， decreased protein expression level of CREB （P<0.05）， and severe renal pathological damage. Compared with the model group， the low-， medium-， and high-dose modified Shenqiwan groups and the irbesartan group showed varying degrees of decreases in blood glucose levels， UACR levels， and the protein expression levels of GSK-3β， TGF-β₁， E-cadherin， Vimentin， FN， PAI-1， and Coll Ⅳ in the kidneys （P<0.05）， increased expression level of CREB protein （P<0.05）， and improved renal pathological damage. ConclusionModified Shenqiwan can effectively reduce blood glucose levels， improve renal function， and alleviate fibrosis， and the mechanism of action is related to the inhibition of the GSK-3β/CREB signaling pathway.