Meta-analysis of the efficacy of immune checkpoint inhibitors in the treatment of NSCLC with different KRAS genotypes / 中国药房

OBJECTIVE To evaluate the efficacy of immune checkpoint inhibitors （ICIs） in the treatment of non-small cell lung cancer （NSCLC） with different KRAS genotypes. METHODS Retrieved from PubMed， the Cochrane Library， Web of Science， Embase， CNKI， Wanfang data and VIP， randomized controlled trials （RCTs） about ICIs alone， combined use of various ICIs or ICIs combined with traditional chemotherapy （trial group） versus traditional chemotherapy （control group） for NSCLC were collected from the inception of the databases to April 1， 2023. After screening literature， extracting data and evaluating quality， meta-analysis， sensitivity analysis and publication bias analysis were conducted by using RevMan 5.4 software. RESULTS A total of 7 RCTs involving 5 980 patients were included. The results of the meta-analysis showed that overall survival （OS） [HR＝ 0.79， 95%CI （0.72， 0.87）， P＜0.000 01] and progression-free survival （PFS） [HR＝0.63， 95%CI （0.50， 0.80）， P＝0.000 2] of trial group were significantly longer than those of control group； furthermore， the OS of KRAS mutant type [HR＝0.63， 95%CI （0.53， 0.75）， P＜0.000 01] and KRAS wild type [HR＝0.87， 95%CI （0.78， 0.98）， P＝0.02]， PFS of KRAS mutant type [HR＝ 0.58， 95%CI （0.43， 0.78）， P＝0.000 3] and KRAS wild type [HR＝0.68， 95%CI （0.47， 0.99）， P＝0.04] in the trial group were all significantly longer than in the control group. Subgroup analysis by different treatment regimens showed that the OS of KRAS mutant type patients receiving first- and second-line treatment regimens， using ICIs alone and those receiving ICIs combined with traditional chemotherapy as well as PFS of KRAS mutant type and wild type patients receiving first-line treatment regimens in the trial group were all significantly longer than in the control group （P＜0.05）. Sensitivity analysis results indicated that the findings of this study were robust. Publication bias results showed that the possibility of publication bias in this study was small. CONCLUSIONS ICIs show significant efficacy in NSCLC patients， and NSCLC patients benefit equally regardless of whether KRAS mutations occur.

Comparison of clinical effect and postopertaive incisions between No-touch technique and traditional retractor in treating calcaneal fracture / 中国骨伤

OBJECTIVE@#To compare clinical efficacy of No-touch technique and traditional retractor in treating calcaneal fracture.@*METHODS@#Clinical data of 74 calcaneal fracture patients with closed Sanders typeⅡ to Ⅳ were retrospectively analyzed from July 2019 to June 2021. According to different treatment methods, the patients were divided into No-touch group and conventional group, 37 patinets in each group. In No-touch group, there were 25 males and 12 females, aged from 19 to 70 years old with an average of (42.64±14.16) years old;17 patients were typeⅡ, 14 patinets with type Ⅲ, 6 patients with type Ⅳ according to Sanders fracture classification;three 2.0 mm Kirschner wires were implanted into the talus body, talus neck, and cuboid bone, and the flap was turned upward to expose the operation area. In conventional group, there were 30 males and 7 females, aged from 19 to 67 years old with an average of (41.56±11.38) years old;17 patients with typeⅡ, 12 patients with type Ⅲ, 8 patients with type Ⅳ according to Sanders fracture classification;the operation was completed by exposing the operation area with traditional retractor. Operation time, postoperative incision complications, postoperaive American Orthopedic Foot and Ankle Society (AOFAS) ankle hind foot score at 6 months between two groups were compared.@*RESULTS@#Seventy-four patients were followed up, and follow-up time in No-touch group ranged from 6 to 17 months with an average of(9.57±2.72) months, while in conventional group ranged from 6 to 16 months with an averge of(9.14±2.71) months, and no difference in follow-up between two groups (P>0.05). Operation time in No-touch group (55.67±7.94) min was shorter than that in conventional group (70.16±9.41) min (P<0.05);four patients in No-touch group occurred incision complications, while 8 patients in normal group, and had statistically difference(P<0.05). Daily activities and support, maximum walking distance (block), ground walking, limited degree of flexion, extension and valgus, foot alignment and total score of AOFAS scores in No-touch group was significantly higher than that of conventional group (P<0.05). There were no significant difference in pain degree, abnormal gait and ankle hind foot stability between two groups(P>0.05). According to AOFAS score, 19 patients got excellent result, 16 good and 2 poor in No-touch group;while 9 excellent, 24 good, and 4 poor in conventional group, and no difference between two groups (P>0.05).@*CONCLUSION@#Compared with traditional retractor in treating calcaneal fracture, No-touch technology could significantly shorten operation time, reduce incidence of postopertive complications, while two methods could improve excellent and good rate of ankle joint function recovery after operation.

Vitexin Inhibits Gastric Cancer Growth and Metastasis through HMGB1-mediated Inactivation of the PI3K/ AKT/mTOR/HIF-1α Signaling Pathway

Purpose@#Gastric cancer (GC) has high morbidity and mortality and is a serious threat to public health. The flavonoid compound vitexin is known to exhibit anti-tumor activity. In this study, we explored the therapeutic potential of vitexin in GC and its underlying mechanism. @*Materials and Methods@#The viability, migration, and invasion of GC cells were determined using MTT, scratch wound healing, and transwell assays, respectively. Target molecule expression was determined by western blotting. Tumor growth and liver metastasis were evaluated in vivo using nude mice. Protein expression in the tumor tissues was examined by immunohistochemistry. @*Results@#Vitexin inhibited GC cell viability, migration, invasion, and epithelial-mesenchymal transition (EMT) in a dose-dependent manner. Vitexin treatment led to the inactivation of phosphatidylinositol-3-kinase (PI3K)/AKT/hypoxia-inducible factor-1α (HIF-1α) pathway by repressing HMGB1 expression. Vitexin-mediated inhibition in proliferation, migration, invasion and EMT of GC cells were counteracted by hyper-activation of PI3K/AKT/HIF-1α pathway or HMGB1 overexpression. Finally, vitexin inhibited the xenograft tumor growth and liver metastasis in vivo by suppressing HMGB1 expression. @*Conclusions@#Vitexin inhibited the malignant progression of GC in vitro and in vivo by suppressing HMGB1-mediated activation of PI3K/Akt/HIF-1α signaling pathway. Thus, vitexin may serve as a promising therapeutic agent for the treatment of GC.

The cardiovascular protection of irisin and its research progress in sports field / 生理学报

Irisin is a circulating myokine induced by exercise, which is a cleaved version of fibronectin type III domain containing protein 5 (FNDC5). It can promote the browning of white fat tissue, increase energy consumption, and decrease weight. Irisin plays an important role in the regulation of various diseases, such as diabetes and coronary heart disease. Different types of exercise have different effects on irisin level in blood circulation, and moderate exercise can reduce cardiovascular symptoms. In this paper, the cardiovascular protective effect of irisin and its research progress in the field of exercise are reviewed, hoping to provide a new target for the prevention and treatment of cardiovascular diseases.

Synergistic effects of Chuanxiong-Chishao herb-pair on promoting angiogenesis at network pharmacological and pharmacodynamic levels / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the synergistic effects of Chuanxiong-Chishao herb-pair (CCHP) on promoting angiogenesis in silico and in vivo.</p><p><b>METHODS</b>The mechanisms of action of an herb-pair, Chuanxiong-Chishao, were investigated using the network pharmacological and pharmacodynamic strategies involving computational drug target prediction and network analysis, and experimental validation. A set of network pharmacology methods were created to study the herbs in the context of targets and diseases networks, including prediction of target profiles and pharmacological actions of main active compounds in Chuanxiong and Chishao. Furthermore, the therapeutic effects and putative molecular mechanisms of Chuanxiong-Chishao actions were experimentally validated in a chemical-induced vascular insuffificiency model of transgenic zebrafifish in vivo. The mRNA expression of the predicted targets were further analyzed by real-time polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The computational prediction results found that the compounds in Chuanxiong have antithrombotic, antihypertensive, antiarrhythmic, and antiatherosclerotic activities, which were closely related to protecting against hypoxic-ischemic encephalopathy, ischemic stroke, myocardial infarction and heart failure. In addition, compounds in Chishao were found to participate in anti-inflflammatory effect and analgesics. Particularly, estrogen receptor α (ESRα) and hypoxia-inducible factor 1-α (HIF-1α) were the most important potential protein targets in the predicted results. In vivo experimental validation showed that post-treatment of tetramethylpyrazine hydrochloride (TMP•HCl) and paeoniflorin (PF) promoted the regeneration of new blood vessels in zebrafifish involving up-regulating ESRα mRNA expression. Co-treatment of TMP•HCl and PF could enhance the vessel sprouting in chemical-induced vascular insuffificiency zebrafifish at the optimal compatibility proportion of PF 10 μmol/L with TMP•HCl 1 μmol/L.</p><p><b>CONCLUSIONS</b>The network pharmacological strategies combining drug target prediction and network analysis identified some putative targets of CCHP. Moreover, the transgenic zebrafifish experiments demonstrated that the Chuanxiong-Chishao combination synergistically promoted angiogenic activity, probably involving ESRα signaling pathway.</p>

Effects of Flow Components C6 and C7 in N-butyl Alcohol Extract from the Leaves of Cestrum nocturnum on the Proliferation and Apoptosis of Human Gastric Cancer Cell SGC7901 / 中国药房

OBJECTIVE:To study the effects of flow components C6 and C7 in n-butyl alcohol extract from the leaves of Ces-trum Nocturnum(CN)on the proliferation and apoptosis of human gastric cancer cell SGC7901. METHODS:C6 and C7 were ob-tained by using different ratio of chloroform and methanol(1:9,1:7)to the gradient elution of CN leaves. After cultured with 0 (blank control),5,10,20,40,80 μg/ml C6 and C7 for 72 h,inhibitory effect of C6 and C7 on the proliferation of SGC7901 was determined by MTT assay. Inhibitory rate and IC50 were calculated. After SGC7901 were cultured with 10 μg/ml C6 and C7 for 72 h,colony formation assay was utilized to detect the effects of C6 and C7 on the cell colony formation,and the rate of colony for-mation was calculated. In addition,Wright/Giemsa and Hoechst33258/PI staining assay were used to observe the change of cytomor-phology. RESULTS:MTT showed that C6 and C7 had inhibitory effect on the proliferation of SGC7901 to different extent;inhibi-tory rates were 22.1%-80.0% and 19.6%-79.7%,and IC50 were 16.4,18.05 μg/ml,respectively. Compared with blank control group,colony formation rate of C6 and C7 group decreased,with statistical significance(P<0.05). The number of apoptotic cells was more in treatment group than in other groups. CONCLUSIONS:Flow components C6 and C7 in n-butyl alcohol extract from the leaves of CN can inhibit the proliferation of SGC7901 cells and induce the apoptosis of them.

320-detector row CT coronary angiography in pre-operation screening of coronary artery disease in patients with rheumatic left artrioventricular valve disease and atrial fibrillation / 第二军医大学学报

Objective To evaluate the diagnostic accuracy of 320-detector row CT coronary angiography(STSL) in detecting preoperative coronary artery disease (CAD) in patients with atrial fibrillation (AF) and rheumatic left artrioventricular valve disease (RMVD), so as to assess its feasibility in screening CAD before operation for RMVD. Methods Thirty-five RMVD patients with persistent CAD were enrolled in the present study. All the patients underwent both STSL and conventional coronary angiography (CCA) before operation. The relationship between mean heart rate and CT image quality was evaluated by Pearson' s correlation coefficient. The sensitivity, specificity, positive predictive value(PPV) and negative predictive value (NPV) of STSL in diagnosis of stenosis (≥50%) were calculated using the conventional coronary angiography as the reference. Kappa statistics was used to assess the agreement between 320-detector STSL and CCA in detecting CAD. Results The mean heart rate was 94. 7 + 21. 4 beats/min in the 35 patients. There was a significant correlation between the mean heart rate and image quality, especially for middle segment of right coronary artery(r=0. 554, P = 0. 002) and middle segment of the circumflex artery (r=0. 559, P=0. 016). The sensitivity, specificity, PPV and NPV of STSL in diagnosing CAD were 87. 5%, 99. 4%, 82. 4% and 99. 6% in segment-based analysis, 87. 5%, 97. 6%, 82. 4% and 98. 4% in vessel-based analysis, and 84. 6%, 86. 3%, 78. 6% and 90. 5% in patient-based analysis, respectively. The kappa statistics were 0. 843, 0. 828 and 0. 699, respectively. Conclusion STSL has a high accuracy in diagnosing CAD, and it can be used for preoperation screening of patients with AF and RMVD.

Effect of rabeprazole-based triple therapy for associated gastric ulcer caused by the helicobacter pylori infection / 中国基层医药

Objective To evaluate the effects of rabeprazole-based triple therapy for associated gastric ulcer caused by the helicobacter pylori(Hp) infection. Methods 100 patients with associated gastric ulcer caused by the Hp infection were randomly divided into treatment group and control group. The treatment group was given rabeprazole-based triple therapy( rabeprazole 10mg bid, amoxicillin 1.0g bid and clarithromycin 500mg bid), and the control group was given omeprazole-based triple therapy( omeprazole 20mg bid, amoxicillin 1.0g bid and clarithromycin 500mg bid). The two groups were given 7 days treatment. Results The total effective rate of clinical symptoms of treatment group was 96. 0% ,the healing rate of the treatment group was 88.0%, the total effective rate of gastroscopy effect of treatment group was 94.0% ,the Hp clearance rate of treatment group was 92.0% and the Hp eradication rate of treatment group was 90. 0% ,which were higher than the control group,but the difference was not significant( P ＞ 0. 05 ). The two groups had no significant adverse reactions. Conclusion Rabeprazole-based triple therapy for associated gastric ulcer caused by the Hp infection was effective, safe and worth popularization and application.

Morphology and developmental groove of maxillary permanent central incisors in Sichuan people / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the morphology of maxillary permanent central incisors of people in Sichuan so as to provide some guidance for emulational restoration of fixed prosthesis.</p><p><b>METHODS</b>Fifty-eight extracted maxillary permanent central incisors were selected from people in Sichuan. All of the teeth involved in this study were selected according to certain criterions. The length and width of the anatomical crowns, the length and the width of the developmental grooves and the angles formed by the two sides of the developmental grooves were measured.</p><p><b>RESULTS</b>The average length and width of anatomical crowns in Sichuan people were (11.9 +/- 1.3) mm and (8.7 +/- 0.8) mm respectively. And the average length and width of the developmental grooves were (5.7 +/- 0.9) mm and (2.1 +/- 0.5) mm respectively. There were no statistical differences between the length of the mesial and distal developmental grooves and so were the width. The ratio of the length of crowns to that of the developmental grooves was 2.1, while for the width, it was 4.2. The angles of the mesial and distal developmental grooves formed by its two sides were 23 degrees +/- 4.7 degrees and 23 degrees +/- 5.7 degrees respectively.</p><p><b>CONCLUSION</b>The average length and width of maxillary permanent incisors were close to Wang's report, which means the length and width of the maxillary permanent incisors of people in Sichuan were quite close to those of Chinese people. The ratio of the length of anatomical dental crowns to that of the developmental grooves was steady, and so was the ratio of the width of them. The angles of the developmental grooves were quite steady too. These may help us in making fixed prosthesis.</p>

Comparison of properties of Mahuang decoction and Maxing Shigan decoction based on biothermodynamics / 中国中药杂志

<p><b>OBJECTIVE</b>To probe into the objectivity and authenticity of four properties beteen Mahuang decoction and Maxing Shigan decoction from biothermodynamics.</p><p><b>METHOD</b>The power-time curves of growth of Staphylococcus aureus at different concentrations between Mahuang decoction and Maxing Shigan decoction were determined by TAM Air Isothermal Calorimeter. The growth rate constants of promotive and inhibitory actions were calculated. Moreover, the difference of properties beteen Mahuang decoction and Maxing Shigan decoction was analyzed from the point of view of TCM theory.</p><p><b>RESULT</b>Both the Mahuang decoction and Maxing Shigan decoction could inhibit the growth and metabolism of Staphylococcus aureus. The k and Pm were decreased with the mass increase of the decoction. However, inhibitory activity of Mahuang decoction with warm property was weaker than that of Maxing Shigan decoction with cool property. Moreover, Mahuang decoction decreased heat output in growth metabolism more weakly than Maxing Shigan decoction. There was a stable difference between them.</p><p><b>CONCLUSION</b>Studying on biothermodynamics can show the difference of four properties of Traditional Chinese Medicine. So, it provides a new and useful means for the study of the properties of traditional Chinese medicine.</p>

Fingerprints of different samples of processed Rhizoma Coptidi by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To set up the HPLC fingerprints of different samples of processed Rhizoma Coptidi (RC) in order to offer evidence for identifying different processed RC and preliminary discussion on processing mechanism.</p><p><b>METHOD</b>Seven different samples of processed RC were determined by HPLC and the results were analyzed by Hierarchical clustering and similarity evaluation system for chromatographic fingerprint of TCM.</p><p><b>RESULT</b>The characteristic of common peaks for fingerprints of different samples of processed RC is distinct, but there are differences among various processed RC. The different samples of processed RC were classified into two and the result of hierarchical clustering is correlated with the property of traditional Chinese drug.</p><p><b>CONCLUSION</b>It is proved that the method which is convenient, rapid and repeat well can be used to identify different samples of processed RC. The change of property of different processed RC is related with processing adjuvant, the possible mechanism is that new components are produced or the contents of known components have changed due to the adjuvant during processing.</p>

Study of the osteogenic ability of mesenchymal stem cells in vivo / 中国矫形外科杂志

[Objective]To investigate the osteogenic ability in vivo of mesenchymal stem cells(MSCs).[Method]Ten-fifteen ml bone marrow aspirates were harvested from the iliac crest of sheep and enriched for MSCs by density gradient centrifugation over a Percoll cushion(1.073 g/ml).After cultured and proliferated,the tissue-engineered bones were constructed with these cells and seeded onto porous 13-tricalcium phosphate ceramics(13-TCP).Then,the constructs were implanted into left metatarsus defect(21 mm in length)of 8 sheep as an experimental group.Porous ?-TCP composed with bone marrow was implanted into defects of same size and position in 8 sheep as a control group.Sheep were sacrificed in the 6th,12th,and 24th week postoperatively,and the implanted samples were examined by radiograph,histology,and biomechanical analysis.[Result]New bone tissue was observed either radiographically or histologically at the defect area of experimental group as early as the 6th week postoperatively;but was not observed in the control group.Because of the new bone formed in a direct manner without progression through a cartilaginous process intermediately,osteoid tissue,woven bone and lamellar bone,occurred earlier in the experimental group than in the control group,in which the bone defects were repaired in a "creep substitution" manner.At the 24th week,radiographs and biomechanical tests revealed an almost complete repair of the defect in experimental group,but only a partial repair in the control group.[Conclusion]MSCs have good reproductive activity not only in vitro,but also in vivo.The new bone formed in a direct manner without progression through a cartilaginous process intermediately when MSCs were transplanted in vivo.The results demonstrated that MSCs was an excellent seed cells for bone tissue engineering.

Determination of total organic acids and salicylic acid in extract of Radix isatidis / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a quantitative method for determination of the total organic acids and salicyclic acid in the extract of Radix Isatidis.</p><p><b>METHOD</b>The total organic acids were determined by acid-base titration and the salicylic acid was determined by HPLC.</p><p><b>RESULT</b>It was shown that contents of total organic acids and salicylic acid in the extract of Radix Isatidis were 13.0% and 0.22%, respectively.</p><p><b>CONCLUSION</b>The method can control the quality of this extract effectively and accurately.</p>

Effects of different extracts from Radix isatidis on lymphocytes of mice by biothermodynamics / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the different extracts from Radix Isatidis on multiplication of mice lymphocytes.</p><p><b>METHOD</b>Lymphocytes were separated and cultured. Immunological activities of different extracts from Radix Isatidis were studied by thermodynamics and the results were tested by the conventional pharmacological experiments.</p><p><b>RESULT</b>The results showed that the water extract and residue had significant immunological effects while organic solvent extracts had immunological activity to some extent.</p><p><b>CONCLUSION</b>The comparison of immunological activity among the extracts from Radix Isatidis were as follows: residue after extracting > general extract > nBuOH extract > EtOAc extract > CHCl3 extract > P E extract.</p>

Application of the second metatarpophalangeal joint by traction prolong transplant repair the defects in the metacarpophalangeal joint / 中华显微外科杂志

Objective Application of the second metatarpophalangeal joint by traction prolong trans- plant repair the defects in the metacarpophalangeal joint,reconstruct the function of it.Methods By means of the apparatus to prolong finger in advance,then transplant the second metatarpophalangeal joint to recon- struct metacarpophalangeal joint for seven cases of obsolete defects in the metacarpophalangeal joint.Results The average of finger prolong was 2.6 cm,consultation from 1 to 4 years.average 2.5 years,thai the trans- plant joints have all survived and osteal concrescence.Through the criterion Chinese Medical Association,good rate was 85.7%. Conclusion It' s a good method to repair obsolete defects in the metacarpophalangeal joint by transplant traction prolong of the second metatarpophalangeal joint.

Thermodynamic study on antibacterial effect of different extracts from Radix Isatis / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study and analyze the antibacterial effects of different extracts from Radix Isatis.</p><p><b>METHODS</b>Staphylococcus aureus was used as the studied object in the experiment. Antibacterial effects of extracts from Radix Isatis were observed by thermocalrimetry on Staphylococcus aureus, together with common pharmacological experiments.</p><p><b>RESULTS</b>The total extract, ethyl acetate (EtOAc) extract, n-butylalcohol (nBuOH) extract, chloroform (CHCl(3)) extract and petroleum (P.E.) extract had antiviral effects to some extent while the residue after extracting had no antibacterial activity. The potency of antiviral activity among them was as follows: nBuOH extract > EtOAc extract > CHCl(3) extract > total extract > P.E. extract.</p><p><b>CONCLUSION</b>The antibacteriall effects of Radix Isatis were not limited to any active portion, showing that Radix Isatis exerts its antibacterial effects by cooperation of different active fractions in varied ways.</p>

The study of tissue-engineering bone for repair of segmental bone defects / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of tissue-engineering bone on repair of segmental bone defects.</p><p><b>METHODS</b>Segmental bone defect of 21mm was created at sheep left metatarsus, which was then implanted with tissue-engineering bone (the experimental group) and pure porous beta-TCP (the control group) respectively. The bone defect in the blank group was left without treatment. After the sheep were sacrificed at the 1st, 3rd, or 6th month postoperatively, the samples were taken and examined by radiological, histological and biomechanical methods as well as scanning electron microscopy. The sheep in the blank group were sacrificed at the 6th month postoperatively.</p><p><b>RESULTS</b>The osteoid tissue, woven bone and lamellar bone in the defect of the experimental group occurred earlier than in the control group. The new bone formed directly without through a cartilaginous intermediate in the experimental group, while the defect was repaired in a "creep substitution" way in the control group. At the 6th month, radiological and biomechanical tests revealed nearly complete repair of the bone defect of the experimental group, partial repair in the control group and non-healing in the blank group.</p><p><b>CONCLUSIONS</b>Tissue-engineering bone can repair bone defect, accelerating healing and without "creep substitution", which is a good option in repair of critical segmental bone defects. This study set up a basis for clinical applications in the future.</p>

Examination of dynamic changes of different-type collagens in bone fracture healing with a polarized light microscopy / 中国组织工程研究

BACKGROUND:Sirius red is a strong acid anionic dye. Being not-easyto-fade and specific, sirius red becomes the best dye for collagen staining.Collagen is a major component of extracellular matrix and has some specific physiological functions. Through synthesis and reconstruction of collagen, bone fracture repair will be accomplished.OBJECTIVE: Picric acid-Sirius red stained slides were observed under a polarized light microscopy for evaluation the dynamic changes in the ratio of different collagen types and their distributions in bone fracture healing.DESIGN: It was a controlled observation.SETTING: It was conducted in the Department of Orthopedics, Renmin Hospital, Wuhan University; Department of Traumatic Orthopaedics, Tianjin Hospital; Department of Traumatic Orthopaedics, Jishuitan Hospital,Medical Department, Peking University; Tissue Engineering Center of Institute of Basic Medical Sciences, Academy of Military Medical Sciences of Chinese PLAMATERIALS: It was conducted at Tissue Engineering Center of Institute of Basic Medical Sciences, Academy of Military Medical Sciences of Chinese PLA from March 2002 to September 2003. Three healthy adult Chinese sheep, male and in weight from 25 to 35 g, were selected.METHODS: All the animals were anesthesized and sterilized; a transverse osteotomy of the trunk of metatarsus was performed; and the end of fracture was fixed with a six-hole Medoff sliding plate. At the post-operative month 1, 3 and 6, samples were taken from bone fractures. After decalcification with EDTA, they were stained with Picric acid-sirius red, and the types and distribution of collagens were observed under a polarized light microscopy.MAIN OUTCOME MEASURES: Types and distributions of collagens in bone lesion in different period of bone healing were investigated.RESULTS: Three sheep used in this study entered the statistical analysis.①Morphological features of various collagens under a polarized light microscopy postoperatively: Type Ⅰ collagen packed tightly, with a strong refraction and yellow, orange or red thick fibres. Type Ⅱ collagen formed a loose reticulation with fibres exhibiting different colour and a weak refraction. Thin fibres of type Ⅲ collagen with weak refraction and green colour formed a loose reticulation. ②Quantitative studies on various collagens under a polarized light microscopy postoperatively: At postoperative month 1,red or orange fibres (type Ⅰ collagen) were rarely seen in bone fracture,while green fibres (typical of type Ⅲ collagen) were dominant with a disorder pack. At postoperative month 3, red or orange fibres increased significantly and the ratio of type Ⅲ collagen reduced. The collagen fibres assembled regularly. At postoperative month 6, thick yellow-red collagen became dominant and thin green type Ⅲ collagen decreased dramatically and arranged in an obvious oblique, spiral and crossed orientations.CONCLUSION: Picric acid-sirius red stain combined with polarized light microscopy technique is not only capable of identifing type Ⅰ and type Ⅲ collagens in bone fraction, but also can reflect the morphological features,distribution and the ratio of these two type collagens. This approach has the virtues of easiness in operation, strong specificity and high sensitivity.

Repair of Sheep Metatarsus Defects by Using Tissue-engineering Technique / 华中科技大学学报（医学）（英德文版）

Tissue-engineering bone with porous β-tricalcium phosphate (β-TCP) ceramic and autologous bone marrow mesenchymal stem cells (MSC) was constructed and the effect of this composite on healing of segmental bone defects was investigated. 10-15 ml bone marrow aspirates were harvested from the iliac crestof sheep, and enriched for MSC by density gradient centrifugation over a Percoll cushion (1. 073 g/ml). After cultured and proliferated, tissue-engineering bones were constructed with these cells seeded onto porous β-TCP, and then the constructs were implanted in 8 sheep left metatarsus defect (25 mm in length) as experimental group. Porous β-TCP only were implanted to bridge same size and position defects in 8 sheep as control group, and 25 mm segmental bone defects of left metatarsus were left empty in 4 sheep as blank group. Sheep were sacrificed on the 6th, 12th, and 24th week postoperatively and the implants samples were examined by radiograph, histology, and biomechanical test. The 4 sheep in blank group were sacrificed on the 24th week postoperatively. The results showed that new bone tissues were observed either radiographic or histologically at the defects of experimental group as early as 6th week postoperatively, but not in control group, and osteoid tissue, woven bone and lamellar bone occurred earlier than in control group in which the bone defects were repaired in "creep substitution" way, because of the new bone formed in direct manner without progression through a cartilaginous intermediate. At the 24th week, radiographs and biomechanical test revealed an almost complete repair of the defect of experimental group, only partly in control group. The bone defects in blank group were non-healing at the 24th week. It was concluded that engineering bones constructed with porous β-TCP and autologous MSC were capable of repairing segmental bone defects in sheep metatarsus beyond "creep substitution" way and making it healed earlier. Porous β-TCP being constituted with autologous MSC may be a good option in healing critical segmental bonedefects in clinical practice and provide insight for future clinical repair of segmental defect.

Repair of sheep metatarsus defects by using tissue-engineering technique / 华中科技大学学报（医学）（英德文版）

Tissue-engineering bone with porous ,betatricalcium phosphate (3-TCP) ceramic and autologous bone marrow mesenchymal stem cells (MSC) was constructed and the effect of this composite on healing of segmental bone defects was investigated. 10-15 ml bone marrow aspirates were harvested from the iliac crest of sheep, and enriched for MSC by density gradient centrifugation over a Percoll cushion (1. 073 g/ml). After cultured and proliferated, tissue-engineering bones were constructed with these,cellS seeded onto porous f-TCP, and then the constructs were implanted in 8 sheep left metatarsus defect (25 mm in length) as experimental group. Porous ,-TCP only were implanted to bridge same size and position defects in 8 sheep as control group, and 25 mm segmental bone defects of left metatarsus were left empty in 4 sheep as blank group. Sheep were sacrificed on the 6th, 12th, and 24th week postoperatively and the implants samples were examined by radiograph, histology, and biomechanical test. The 4 sheep in blank group were sacrificed on the 24th week postoperatively. The results showed that new bone tissues were observed either radiographic or histologically at the defects of experimental group as early as 6th week postoperatively, but not in control group, and osteoid tissue, woven bone and lamellar bone occurred earlier than in control group in which the bone defects were repaired in "creep substitution" way, because of the new bone formed in direct manner without progression through a cartilaginous intermediate. At the 24th week, radiographs and biomechanical test revealed an almost complete repair of the defect of experimental group, only partly in control group. The bone defects in blank group were non-healing at the 24th week. It was concluded that engineering bones constructed with porous -TCP and autologous MSC were capable of repairing segmental bone defects in sheep metatarsus beyond "creep substitution" way and making it healed earlier. Porous ,-TCP being constituted with autologous MSC may be a good option in healing critical segmental bone defects in clinical practice and provide insight for future clinical repair of segmental defect.