Establishing a model of femoral head necrosis by alcohol burning in Cyan-Shank Partridge Chicken: A feasibility analysis / 中国组织工程研究

BACKGROUND: To date, the pathological mechanism of femoral head necrosis is still unclear, and the existing experimental animal models of femoral head necrosis all have shortcomings. OBJECTIVE: To establish a simple, reliable and stable animal model of femoral head necrosis in a Cyan-Shank Partridge Chicken, with similar biomechanical characteristics to human beings. METHODS: Twenty-four adult young Cyan-Shank Partridge Chickens were randomly divided into experimental group and control group (n=12 per group). One side of the femoral head was randomly selected for operation, and the other side was untreated (untreated group). In the experimental group, alcohol was used to burn the bone tissue in the drilled hole followed by implantation of the original bone. In the control group, the original bone was directly re-implanted after drilling. X-ray and CT scan examinations were performed 1 month after operation. The femoral head was then dissected for general observation, microCT observation and pathological staining. The study protocol was approved by the Animal Ethic Committee of Xuzhou Medical University in China (approval No. AEC2018-020). RESULTS AND CONCLUSION: All the Cyan-Shank Partridge Chickens were alive without wrongful death. They could walk upright with no obvious claudication at 1-3 days after operation. X-ray and CT results revealed collapse of the femoral head with unclear trabecular bone in the experimental group. General observation indicated that the femoral head was collapsed with no smooth surface in the experimental group. MicroCT results showed significantly reduced number of bone trabeculae and significantly increased trabecular thickness, trabecular spacing and bone volume fraction in the experimental group as compared with the control and untreated groups (P < 0.05). Pathological staining showed that there was no obvious necrosis but similar shape of the femoral head in the control and untreated groups, but the number of subchondral vessels was less in the control group than the untreated group. However, the cells arranged disorderly and obvious necrosis such as empty lacuna was observed in the experimental group. To conclude, the use of alcohol to burn the bone tissue of the femoral head of the Cyan-Shank Partridge Chicken can be used to establish a typical animal model of femoral head necrosis, which has similar pathological characteristics of human femoral head necrosis.

Effect of Fructus Mume Extract on Repair of Renal Injury Induced by Nanobacteria in Rats and Its Relationship with Antagonistic Stone Formation / 中国医科大学学报

Objective To observe the early effect of Fructus mume extract on KIM-1 and OPN levels in rats with kidney stone formation induced by nanobacteria and to investigate the therapeutic significance of F.mume extract on early kidney stone formation.Methods Nanobacteria were separated and cultured from human upper urinary calculi.The study group appropriately included 54 healthy male SD rats.The renal calculus model was constructed by tail vein injection of nanobacteria.A kidney stone model was created with an F.mume extract intervention,and rats were killed at different stages of the intervention.Real-time polymerase chain reaction was used to detect the KIM-1 mRNA expression in rat renal tissue,and the KIM-1 concentration in urine was detected by using enzyme-linked immunosorbent assay.We detected kidney tissue stone crystals and OPN expression by using hematoxylin-eosin staining and immunohistochemistry.Results Renal tubules of the experimental model were significantly expanded,that is,the formation of renal tubular stones.The early KIM-1 and OPN expression levels were increased.The above-mentioned changes positively correlated with the injection time of the nanobacteria,and F.mume extract antagonized the changes.Conclusion F.mume extract may be useful for the repair of renal injury to reduce kidney stone formation,which may be related to the gene regulation of KIM-1 and OPN.

Evaluation of combination therapy with M and α receptors antagonist for the treatment of double-J stent related lower urinary tract symptoms / 中华泌尿外科杂志

Objective To evaluate the efficacy of combination therapy with M and α receptors antagonist for the treatment of double-J stent related lower urinary tract symptoms.Methods From May 2013 to May 2014,131 cases,including 71 male and 60 female cases,were accepted the doubte-J stent indwelling after the ureteral lithotripsy,laparoscopic ureterlithotomy and pyeloureteroplasty.Their data was retrospectively reviewed.The age ranged from 29 to 64 years old,mean (47.4 ± 15.2) years old.They were divided into 4 groups randomly,including group A (control group,n =30),no drugs were taken;group B (tamsulosin group,n =34),0.2 mg tamsulosin was taken qd;group C (solifenacin group,n =32),5 mg solifenacin was taken qd;group D (tamsulosin combined with solifenacin group,n =35),0.2 mg tamsulosin and 5mg solifenacin were taken qd.The IPSS scores,QOL scores and visual analogue pain scale (VAPS) scores were assessed pre-operation,1 week after operation,and 4 weeks after operation,respectively.Results All patients were followed-up until the end of this study.In each time point,the IPSS scores in group A was 9.01 ± 2.79,13.18 ± 3.79 and 13.79 ± 3.76,respectively.In group B,the IPSS scores were 7.89 ± 4.29,12.39 ±3.90 and 12.21 ±3.87,respectively.In group C,the IPSS scores were 7.94 ±4.27,12.70 ±4.01 and 11.98 ±4.69,respectively.In group D,the IPSS scores were 8.21 ±3.18,11.97 ±5.03 and 8.17 ± 3.25,respectively.Significant difference in total IPSS scores and obstruction symptom scores were shown between pre-and post-operation (P ＜ 0.05).Comparing to other groups,group D exhibited the significant improvement in IPSS scores 1 and 4 weeks after the operation (P ＜0.05).4 weeks after operation,the QOL scores in group D was significantly lower than that in other groups (P ＜ 0.05).While the VAPS scores didn t show significant differences among those groups (P ＞ 0.05).Conclusion M and α receptors antagonist combination therapy can significantly improve lower urinary tract symptom due to indwelling double J stents.

Inhibitory effect of Genipin on uncoupling protein-2 and energy metabolism of androgen-independent prostate cancer cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore whether the inhibitory effect of Genipin on uncoupling protein-2 (UCP-2) in mitochondria is involved in energy metabolism of androgen-independent PC3 prostate cancer cells.</p><p><b>METHODS</b>PC3 prostate cancer cells were cultured and treated with Genipin at the concentrations of 40, 80, and 160 μmol/L for 48 hours. Then the proliferation of the cells was detected by MTT assay, the expression of UCP-2 mRNA determined by RT-PCR, and the content of intracellular pyruvic acid (PA) and the activity of succinate dehydrogenase (SDH) in the mitochondria measured by visible spectrophotometry.</p><p><b>RESULTS</b>With the increased concentration of Genipin, the proliferative activity of the PC-3 cells, the expression level of UCP-2 mRNA, the content of intracellular PA and the activity of SDH in the cells were all decreased, namely, with the enhanced inhibitory effect of Genipin on UCP-2, a trend of reduction was observed in the proliferation of the cells, intracellular PA content, and SDH activity in the mitochondria.</p><p><b>CONCLUSION</b>Genipin is involved in the energy metabolism of androgen-independent PC3 prostate cancer cells by reducing the content of intracellular PA and the activity of SDH in the mitochondria, which may be associated with its inhibitory effect on UCP-2.</p>

Effects of chronic arsenic poisoning on serum inhibin-B and testosterone in male rats / 中华地方病学杂志

Objective To evaluate the effects of chronic arsenic poisoning on serum inhibin-B (INH-B) and testosterone of male rats.Methods Forty Sprague-Dawley rats in cleanliness grad were randomly divided into four groups according to body weight:high-dose,middle-dose,low-dose arsenic exposure groups and control group,10 animals in each group.Rats in high-dose,middle-dose,and low-dose groups were administered with arsenic in concentrations of 10.0,5.0,2.5,0.0 mg/kg in distilled water,respectively.Rats in control group were fed with distilled water freely.After 9 months,serum INH-B and testosterone were tested using an ELISA Kit.Results Serum INH-B and testosterone levels in the above mentioned four groups,respectively,were (66.31 ± 37.21),(75.13 ± 29.88),(119.02 ± 36.10),(133.04 ± 38.67)ng/L and (1.292 ± 0.715),(1.757 ± 0.649),(2.359 ± 0.675),(2.817 ± 0.890)μg/L,and the differences between different groups were statistically significant (F=84.88,8.214,all P ＜ 0.05).Serum INH-B and testosterone levels in high-dose and middle-dose groups were significantly lower than that of control group(all P＜ 0.05),whereas no significant difference was found between low-dose group and control group (all P ＞ 0.05).Conclusion Chronic arsenic poisoning has led to decreased concentration of serum INH-B and testosterone in male rats.

STC-1 is involved in anti-hypoxia proliferative balance of renal cancer cells by down-regulation of intracellular Ca2+ and HIF-1α levels / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate effects of stanniocalcin-1 (STC-1) on proliferation balance under hypoxic condition in renal cancer cells and its mechanism.</p><p><b>METHODS</b>Hypoxic model was induced on renal cancer GRC-1 cells (Group H), the cells were treated with STC-1 protein at concentrations of 0.1 nmol/L (H1), 0.5 nmol/L (H2), 1.0 nmol/L (H3), or normal saline (H0) for 48 h， respectively. Cells proliferation was measured by MTT assay; mRNA and protein expressions of hypoxia inducible factor 1α (HIF-1α) and STC-1 in GRC-1 cells were detected by RT-PCR and ELISA, respectively; the intracellular levels of Ca2+ and adenosine triphosphate (ATP) were determined by fluorescence spectrophotometry and spectrophotometry, respectively.</p><p><b>RESULTS</b>The expression of HIF-1α, STC-1 and Ca2+ levels were increased in GRC-1 cells under hypoxia condition; STC-1 reversed these changes in a dose-effect manner. Hypoxia significantly inhibited cell proliferation and the generation of ATP in GRC-1 cells and exogenous STC-1 reversed the effects of hypoxia; ATP generation increased gradually with increasing STC-1 concentration, but the cell proliferation was reduced.</p><p><b>CONCLUSION</b>Exogenous STC-1 can promote the proliferation of renal cancer cells in hypoxia condition by reducing HIF-1α expression and Ca2+ content and increased ATP production, but the progressive inhibition of HIF-1 α hindered the renal carcinoma cell proliferation further, which indicates that STC-1 may be involved in anti-hypoxia proliferative balance of renal cancer cells.</p>

Effects of stanniocalcin-1 and hypoxia-inducible factor-1α on mitochondrial membrane potential stability in renal carcinoma cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the effects of stanniocalcin-1 (STC-1) and hypoxia-inducible factor-1α (HIF-1α) on the calcium and thus on the mitochondrial membrane potential (Δψm) in renal carcinoma cells.</p><p><b>METHODS</b>We successfully established the renal carcinoma cell models with high HIF-1α gene expression. After various concentrations of STC-1 solutions were added to the culture medium, the proliferation of cells, expressions of HIF-1α and STC-1, levels of Ca(2+), Δψm, and mPTP were detected by MTT, RT-PCR, ELISA, fluorescence spectrophotometry, and ultraviolet spectrophotometry, respectively.</p><p><b>RESULTS</b>The proliferation of renal carcinoma cells and Δψm were improved after HIF-1α gene transfection, STC-1 protein intervention, and STC-1 protein intervention after gene transfection. While the intracellular Ca(2+) level and mPTP were decreased significantly (P<0.05), all the changes were intensified with the gradual increase of STC-1. However, the increasing trend of cell proliferation gradually declined.</p><p><b>CONCLUSION</b>HIF-1α may participate in malignant proliferation of renal carcinoma cells by promoting STC-1 proliferation or down-regulating Ca(2+); however, such an effect may be gradually attenuated due to the inhibitory effect of STC-1 on HIF-1α.</p>

Role of stanniocalcin 1 in brain injury of coal-burning-borne fluorosis rats / 中国地方病学杂志

Objective To observe the change of stanniocalcin 1 (STC1) and calcium content in brain of coal-burning-borne fluorosis rats,and to explore the role of STC1 in brain injury of coal-burning-borne fluorosis.Methods Twenty four male SD rats were randomly divided into control,low,medium,and high fluoride groups according to body mass.Control group was fed conventional rat chow(fluorinated 1.3 mg/kg),and low,medium and high fluoride groups fed with fluorinated feed(20.0,40.0,60.0 mg/kg).All rats were given distilled water and feed ad libitum.One hundred and eighty days after modeling,STC1 protein and gene expression in the brain tissue of rats were detected using immunohistochemistry and RT-PCR and calcium content of brain tissue was detected.Results The cell positive rates of STC1 in low,medium,high fluoride groups [(48.10 + 2.11)％,(54.90 ± 1.73)％,(79.30 ± 3.71)％] were significantly higher than that of the control group[(24.70 + 3.53)％,all P ＜ 0.05],the cell positive rate of high fluoride group was significantly higher than that of the low and medium fluoride groups (all P ＜ 0.05).The STC1 mRNA expression of low,medium and high fluoride groups (0.58 ± 0.09,0.85 ± 0.17,1.75 ± 0.04) were significantly higher than that in the control group(0.37 ± 0.12,all P＜ 0.05),the STC1 mRNA expressions of high fluoride group was significantly higher than that of the low and medium fluoride groups (all P ＜ 0.05).The brain cortex calcium ion concentrations of low,medium and high fluoride groups[(138.62 + 4.19),(167.43 + 6.57),(189.45 + 3.72)nmol/L] were significantly higher than that in the control group [(101.47 + 9.46)nmol/L,all P ＜ 0.05],the brain cortex calcium ion concentrations of high fluoride group was significantly higher than that of the low and medium fluoride groups(all P ＜ 0.05),and the medium fluoride groups was higher than the low groups (P ＜ 0.05).Conclusion STC 1 may be involved in brain damage of coal-burning-borne fluorosis rats through regulating calcium balance.

Experience of carrying out the second class of biotechnological pharmaceutics / 中华医学教育探索杂志

One of the aims of biotechnological pharmaceutics is to educate innovative highquality medical talents.The second classroom activities can be used as a complementation to ensure the teachiug effectiveness of the course.This study focuses on the experience of carrying out the second class of biotechnological pharmaceutics and tends to provide references for teachers who are going to carry out the second class.

Effect of endemic fluoride poisoning caused by coal burning on the oxidative stress in rat testis / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the effect of endemic fluoride poisoning caused by coal burning on the oxidative stress in rat testis.</p><p><b>METHODS</b>Totally 40 male SD rats were equally randomized into four groups control group, low fluorosis group, middle fluorosis group, and high fluorosis group. Rats in all three fluorosis groups were fed with corn dried by burning coal obtained from endemic fluorosis areas with high fluoride, and thus the animal models of fluorosis were established. After 120 and 180 days, all the rats were sacrificed. Testis tissues were stained with hematoxylin eosin and observed under light microscope. The malonaldehyde (MDA) content, superoxide dismutase (SOD) activity, total nitric oxide synthase (TNOS), and inducible nitric oxidase synthase (iNOS) were measured by biochemical methods in the testis tissues. The content of NaF in testis was measured by fluorine selective electrode.</p><p><b>RESULTS</b>The rat fluorosis models were successfully established. The fluoride content in testis was significantly increased in all the fluorosis groups(P<0.01). Testicular structures were damaged in all of fluoride groups. The TNOS, iNOS activities, and MDA content of each fluoride group were significantly higher than that of the control group on day 120 and 180 (P<0.05 or 0.01 ). The TNOS, iNOS activities, and MDA content significantly increased in a dose dependent manner (P<0.05 or 0.01). The SOD activities significantly decreased in all the fluoride groups (P<0.05 or 0.01).</p><p><b>CONCLUSIONS</b>Endemic fluoride poisoning caused by coal burning can cause disorders in the oxidative system and antioxidative system in rat testis. The oxidative stress may play an important role in the fluorides induced reproductive toxicity in male rats.</p>

Dynamic observation on the bladder acellular matrix grafts for substituting albuginea penis in rabbits / 中国组织工程研究

BACKGROUND: At present, bladder acellular matrix grafts have been successfully used for substituting animal bladder and urinary canal, and for repairing hypospadia. However, reports on bladder acellular matrix grafts for substituting albuginea penis need to be investigated. OBJECTIVE: Allogeneic bladder acellular grafts were used for substituting albuginea penis of rabbits, in order to observe repairing results. DESIGN: A randomized controlled observation. SETTING: West China Medical Laboratory Animal Center and West China Laboratory of Tissue Engineering of Sichuan University as well as Laboratory of Tissue Engineering of Guiyang Medical College. MATERIALS: Fifty male healthy New Zealand Rabbits of grade 3, weighing 2.6-3.0 kg, without phimosis and penis dysplasia, and without presence of phallocampsis after normal saline being perfused, were provided by Huaxi Laboratory Animal Center of Sichuan University. METHODS: This study was performed at the West China Laboratory Animal Center and West China Laboratory of Tissue Engineering of Sichuan University as well as Laboratory of Tissue Engineering of Guiyang Medical College between December 2005 and June 2007. Bladders were taken from 10 experimental rabbits for preparing bladder acellular matrix grafts. The other 40 New Zealand rabbits were randomly divided into the control group, and the bladder acellular matrix grafts group, with 20 in each. An area of 10 mm×5 mm of albuginea penis was resected from dorsum penis of each rabbit. Suture in situ of albuginea penis and bladder acellular matrix grafting were conducted in rabbits of the control group and bladder acellular matrix grafts group, respectively. In the 2nd, 6th, 12th and 24th weeks postoperatively, each rabbit was intracavernously perfused normal saline for inducing penile erection, separately, in order to observe phallocampsis. At above-mentioned each time point, experimental animals were sacrificed. Sample was taken from surgical region for haematoxylin-eosin (HE) staining and Masson trichrome staining, in order to observe the changes of tissue and structure of surgical region. Types Ⅰand Ⅲ collagen fiber areas were detected by Stirus red staining, and the expressions of inducible nitric oxide synthase(iNOS) and transforming growth factor-β1 (TGF-β1) were detected by immunohistochemical staining. MAIN OUTCOME MEASURES: ①Phallocampsis status. ② Changes of tissue and structure of surgical region. ③iNOS and TGF-β1 expressions. ④TypeⅠand Ⅲ collagen fiber areas.RESULTS: Forty experimental rabbits were involved in the penile surgery, two of them died from overdose anesthesia, two died from chordapsus, so the remaining thirty-six rabbits were involved in the final analysis. In the 6th week postoperatively, phallocampsis reached its highest level, and 2 rabbits in the control group and 1 rabbit in the bladder acellular matrix grafts group presented phallocampsis. In the 12th week, every rabbit presented phallocampsis. In the 24th week, 1 rabbit in the control group but none in the bladder acellular matrix grafts group presented phallocampsis. In the 2nd week, the structure of surgical regions of each rabbit was poorly clear, with remarkable inflammatory infiltration. In the bladder acellular matrix grafts group, grafting regions presented cells ingrowing the bladder acellular matrix grafts. Masson trichrome staining results showed that in the surgical region, tunica albuginea fibers were thin and poorly arranged. In the 6th week, tunica albuginea recovered its integrity, and bladder acellular matrix grafts could not be distinguished. No significant difference existed between two groups. In the 24th week, tunica albuginea was even and complete in the sugical region, and fibers restored their arrangement of circular muscle in inner layer and longitudinal muscle in outer layer, without difference from normal tunica albuginea. iNOS and TGF-β1 expressions were the strongest in the 2nd week, and they were found in the fibrocytes and vascular endothelial cells in the 6th week, but a little in the 12th and 24th weeks postoperatively. There were no remarkable differences in iNOS and TGF-β1 expressions between two groups at the same time point. In the 2nd week, typesⅠand Ⅲ collagen fibers co-existed with equivalent proportion. Then, typeⅠcollagen fibers were gradually increased, while type Ⅲ collagen fibers were on the contrary. In the 24th week, typeⅠcollagen fibers took the main place and type Ⅲ collagen fibers were unremarkable. CONCLUSION: Bladder acellular matrix grafts have no remarkable inflammatory reactions and fibrosis in repairing tunica albuginea of New Zealand rabbits, so they are very ideal grafting materials for penile surgery.

Therapeutic effect of agonistic CD40 monoclonal antibody combined with CTL on hu-SCID mouse B lymphoma model / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the therapeutic effect of agonistic CD40 monoclonal antibody combined with tumor specific cytotoxic T lymphocyte (CTL) on B lymphoma.</p><p><b>METHODS</b>Human B lymphoma cell line, Daudi cells, were cultured with CD40 mAb (5C11) for 24 and 48 hours, respectively. Annexin V/PI-binding assay was employed to analyze apoptosis, and FCM to analyze Fas (CD95) expression. Human peripheral monocyte-derived DC were loaded with apoptotic Daudi cells and stimulated by SC11 for further maturation. Tumor specific CTL were generated in vitro by co-culture of mature DC with autologous T lymphocytes. DNA fragmentations of Daudi cells treated with 5C11, CTL or 5C11 combined with CTL were determined by JAM assay. To establish the B lymphoma model, Daudi cells were subcutaneously injected into humanized SCID mice (hu-SCID). 1 or 3 weeks after tumor transfer. tumor-bearing mice were respectively treated with SC11, CTL, 5C11 combined with CTL by intraperitoneal injection. Tumor volume in differently treated mice was measured every week after therapy, and the survival of tumor-bearing mice was recorded.</p><p><b>RESULTS</b>5C11 significantly up-regulated FAS expression in Daudi cells, but had no significant effect on apoptosis rate of Daudi cells. Tumor-specific CTL could effectively kill Daudi cells. Fragmentation of Daudi cells co-cultured with CTL was remarkably enhanced by combination with SC11. Tumor growth in hu-SCID mice was apparently delayed by treatment with SC11, CTL, or SC11 combined with CTL. Moreover, minimal tumor burden mice got 30.0% or 70.0% complete remission (CR), respectively, when received CTL treatment or combination treatment of SC11 with CTL, and the lifespan of tumor bearing mice was also prolonged significantly.</p><p><b>CONCLUSION</b>SC11 may enhance the sensitivity of Daudi cells to apoptosis by up-regulation of Fas expression and promote cytotoxicity of CTL in vitro and therapeutic effect in vivo.</p>

Pathological changes of testis tissue in SARS patients / 基础医学与临床

Ojective To study the pathological changes of testis tissue in SARS patients.Methods Tissue specimens were studied by HE staining、TUNEL and immunohistochemistry(IHC).Results SARS patients showed that widespead germ cells destruction,few or no spermatozoon in the seminiferous epithelium and the lumen,thickened basement membrane、 peritubular fibrosis、 vascular congestion and leukocytes infiltration.The apoptotic seminiferous cells increased significantly(P

Development of a molecular screening test for hereditary hearing loss and genetic susceptibility to aminoglycoside toxicity for Chinese population / 北京大学学报(医学版)

Objective: To develop a molecular screening test for genetic defects on hearing loss related genes has significant impacts on early identification of hereditary hearing loss and genetic susceptibility to aminoglycoside ototoxicity. Early identification of pre-lingual hearing loss is very important for patient's language development, academic achievement, and social skill. Two common mutations, the 235delC in GJB2 gene and the mutation A1555G in mitochondrial DNA, are included in the newly developed screening panel for Chinese population. Methods: A molecular genetic assay, based on fluorescent labeled multiplex PCR and automatic DNA fragment analyzing techniques, was developed to detect both mutations simultaneously. Results: This assay was able to detect both mutations from patient's samples, and pooled DNA tests, as well as suitable to detect mutation from the DNA extracted from dried blood spot and buccal swab. Conclusion: This assay could be a useful tool for newborn screening and carrier screening for the hereditary hearing loss for the Chinese population.

Lung pathology and pathogenesis of severe acute respiratory syndrome: a report of six full autopsies / 中华病理学杂志

<p><b>OBJECTIVE</b>Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first manifested in humans in November 2002. The SARS-associated coronavirus (SARS-CoV) has been identified as the causal agent, but the pathology and pathogenesis are still not quite clear.</p><p><b>METHODS</b>Post-mortem lung samples from six patients who died from SARS from April to July 2003 were studied by light and electron microscopy, Masson trichromal staining and immunohistochemistry. Evidence of infection with the SARS-CoV was determined by reverse-transcription PCR (RT-PCR) , serological examination and electron microscopy.</p><p><b>RESULTS</b>Four of six patients had serological and RT-PCR evidence of recent infection of SARS-CoV. Morphologic changes are summarized as follows: (1) Diffuse and bilateral lung consolidation was seen in all patients (6/6) with increasing lung weight. (2) Diffuse alveolar damage was universal (6/6) with hyaline membrane formation (6/6), intra-alveolar edema/hemorrhage (6/6), fibrin deposition (6/6), pneumocyte desquamation (6/6). A marked disruption in the integrity of the alveolar epithelium was confirmed by immunostaining for the epithelial marker AE1/AE3 (6/6). (3) Type II pneumocytes, with mild hyperplasia, atypia, cytomegaly with granular amphophilic cytoplasm and intracytoplasmic lipid accumulation (5/6). (4) Giant cells in the alveoli were seen in five of 6 patients (5/6) , most of which were positive for the epithelial marker AE1/AE3 (5/6), but some cells were positive for the macrophage marker CD68(2/6). (5) A pronounced increase of macrophages were seen in the alveoli and the interstitium of the lung (6/6), which was confirmed by histological study and immunohistochemistry. (6) Haemophagocytosis was present in five of the 6 patients(5/6). (7) Lung fibrosis was seen in five patients(5/6), with alveolar septa and interstitium thickening(5/6), intraalveolar organizing exudates (6/6) and pleura thickening (4/6). Proliferation of collagen was confirmed by Masson trichromal staining, most of which was type III collagen by immunostaining. The formation of distinctive fibroblast/myofibroblast foci was seen in five patients (5/6) by light microscopy and immunochemistry. (8) Squamous metaplasia of bronchial mucosa was seen in five patients(5/6). (9) Thrombi was seen in all patients(6/6). (10) Accompanying infection was present in two patients, one was bacteria, the other was fungus. In addition, electron microscopy revealed viral particles in the cytoplasm of alveolar epithelial cells and endothelial cells corresponding to coronavirus.</p><p><b>CONCLUSION</b>Direct injury of SARS-CoV on alveolar epithelium, prominent macrophage infiltration and distinctive fibroblast/myofibroblast proliferation may play major roles in the pathogenesis of SARS.</p>

Effect of agonist anti-CD40 mAb 5C11 on the induction and biological characteristics of leukemic dendritic cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the impact of an agonist anti-CD(40) monoclonal antibody 5C11 on the induction and biological characteristics of leukemic dendritic cells.</p><p><b>METHODS</b>Combinations of 5C11 and different cytokines were used to induce differentiation of leukemic blasts into dendritic cells. Morphology was observed by light microscopy. Surface antigens of the induced cells were analyzed by fluorescence-activated cell sorting (FACS), the yields of dendritic cell by cell counting, the levels of IL-6 and IL-12 by ELISA, T cell proliferating activity by allo-mixed lymphocyte reaction (MLR) in vitro. Allogeneic T cells were stimulated with leukemic dendritic cells and T-cell cytotoxicity was measured by MTT assay.</p><p><b>RESULTS</b>When cultured with combinations of 5C11 and different cytokines, the leukemic cells isolated from the patients could differentiate into dendritic cells. The morphology showed typical features of dendritic cells, which expressed high levels of CD(40), CD(80) and CD(86). In comparison with the original leukemia cells, the leukemic dendritic cells secreted less IL-6 but more IL-12 (P < 0.05). The leukemic dendritic cells were potent to stimulate the proliferation of allogeneic T cells, and the latter was able to lyse the original leukemia cells.</p><p><b>CONCLUSION</b>Leukemic blasts could be induced to differentiate into functional dendritic cells. It may be of great value in the adoptive immunologic therapy of leukemia.</p>

Gene detection of severe acute respiratory syndrome-related coronavirus / 中华病理学杂志

<p><b>OBJECTIVE</b>To develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood.</p><p><b>METHODS</b>A pair of primers and a probe (molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank (G130027616).</p><p><b>RESULTS</b>In the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process. The 145 bp fragment of PCR product was further confirmed by conventional PCR assay and proved by DNA sequencing to be identical to the target sequence to which the probe was hybridized.</p><p><b>CONCLUSION</b>This assay has a broad application for clinical diagnosis and surveillance investigation.</p>