Objective To investigate the effect of predeposit autotransfusion in operation of the patients with lumbar disc protrusion.Methods Fifty patients of transfusion with lumbar disc protrusion were assigned into two groups by stratified sampling randomly,30 patients whose blood were predeposited before operation in experimental group,and the other 20 patients whose blood were not predeposited before operation in control group.The blood loss,the blood requirements during operations,the hemotological routine indexes and the complications related to blood transfusion were compared respectively.Results The blood loss of experimental group [ (720 ± 665 ) ml ] perioperative period was lower than that of control group [ ( 1060 ± 558 ) ml ],but there was no significant difference between two groups (P ＞ 0.05 ).All the patients in experimental group went through perioperative period safely without allogenic blood transfusion.Hemoglobin,red blood cell and white blood cell were not significantly different between two groups before and after operation for 3,7 days (P＞ 0.05 ),the platelet count after operation for 7 days was significantly different between two groups (P ＜ 0.05).No complication was observed in experimental group but 1 case with complication was observed in control group.Conclusions Predeposit autotransfusion is an effective to avoid homologous blood transfusion and its complications for the patients with lumbar disc protrusion.Furthermore,the clinical effect is not significantly different between the predeposit autotransfusion patients and the allogenic blood transfusion patients.

ObjectiveTo investigate and evaluate the result and the possibility of the clinical application of autologous chondrocyte implant (ACI).MethodsFrom November 2007 to June 2009,6 cases of knee articular cartilage defect were treated with ACI,including 2 males and 4 females with an average age of 39.5 years (range,19-55).All the defects were located on the condyles of femur with a mean size of 7.3 cm2 (range,3.8-11.6).ACI comprises a two-stage procedure:chondrocytes are first harvested from the non-load bearing area of the joint,expand in vitro to acquire enough cells,and then the chondrocytes are implanted.The defect of cartilage were covered with bone membrane and fixed with sutures and fibrin albumen glue.Lysholm score system,International Knee Documentation Committee (IKDC) grading system,and MRI were used to evaluate the effect of ACI,6 and 12 months post-operatively.ResultsAll the patients were followed up.The clinical outcomes of the 6 and 12 months follow-ups demonstrated increased of clinical scores.The MRI follow-up showed good filling of the defect with tissue having the imaging appearance of cartilage in all patients.Only one patient suffered adhesion,because she refused to finish rehabilitation exercises as our treatment advises.ConclusionAs the clinical effect of ACI for knee cartilage defect is satisfied,the ACI may be a good choice for treating knee cartilage defect in future.It is very important to control the indications strictly and guarantee to finish the post-operative rehabilitation exercises.

Objeetive To discuss the correlation of creatinine reduction ratio(CRR2)from posttransplant day 1 to day 2 and early graft function recovery status after kidney transplantation. Methods Clinical data of 80 patients after renal transplantation from Jan 2005 to Mar 2007 were retrospectively analyzed.Patients were divided into three groups according to the post-operative serum creatinine level:53 patients within IGF group[cereatinine＜265.2 μmol/L by post-operative day(POD)no.5],14 patients within SGF group(creatinine＞265.2 μmol/L on POD no.5,but no need for dialysis),and 13 patients within DGF group(need for dialysis in the first week post-transplant).Then the value and 99%CI of CRRz of these three groups were calculated. Results The value of CRR2 of IGF,SGF and DGF was(46.8±14.6)%,(25.6±13.5)%and(0.7±17.7)%respectively.And CRR2 99%CI of IGF,SGF and DGF was 41%-52%,15%-36%and-14%-16 0A respectively.There was significant difference in the value of CRR2 among IGF,SGF and DGF group.So a criteria for early diagnosis of IGF,SGF and DGF by CRR2 99%C1 was established:IGF(CRR2≥40%),SGF (15%＜CRR2＜40%)and DGF(CRR2≤15%). Conclusion CRR2 has a good correlation with early graft function recovery after kidney transplantation,and can be used to predict the occurrence of SGF and DGF.

Objective To explore the expression changes and possible molecular mechanisms of Neuritin and HSP60 during the repair of liver injury,and to provide an experimental basis for the study of the repair of liver injury. Methods Forty-eight Sprague-Dawley(SD)rats were randomly divided into control group without any treatment(n=6)and experimental group(n=42)underwent 70% hepatectomy to induce acute liver injury,and 6 h,12 h,24 h,48 h,3 d,7 d and 14 d after the operation,the left lobe resection of the residual liver was per-formed. Immunoblotting technique (Western blot) was used to detect the expression difference of Neuritin and HSP60 in liver tissue of the corresponding time points. Hematoxylin eosin staining(hematoxylin-eosin staining, HE)was used to observe the expression changes at each time point in liver pathology,and tail vein blood to detect ATL and AST changes. Results (1)Compared with those in the control group,the serum ALT and AST levels in the experimental group were increased at 6 h,12 h,24 h and 48 h after the operation,and reached the peak at 48 h postoperatively but those began to decrease 3 d postoperatively and was almost normal 7 d postoperatively. The difference was statistically significant(P < 0.05). Pathological findings:compared with that in the control group, the hepatic lobule structure in the experimental group was disorderly. The obvious balloon like change reached the peak 48 h postoperatively.(2)In the whole process of repair of liver injury,the expression of Neuritin and HSP60 showed differences in the opposite. There was a significantly negative correlation between Neuritin expression and the repair or the aggravation of the injury(P<0.001)and the lowest expression was observed 48 h postoperatively. There was a significantly positive correlation between HSP60 expression and the repair or the aggravation of the injury(P < 0.001)and the highest expression was observed 48 h postoperatively. Conclusions (1)With simple 70% left hepatic lobectomy,the repair of liver injury model of SD rats is successfully established and the heaviest injury is observed 48h postoperatively. With a high success rate(100%,42/42),simple and practical,the method provides a reliable and convenient animal model for the study of liver regeneration,liver injury and liver transplan-tation.(2)The expression of Neuritin decreases gradually after liver injury and reaches the lowest 48 h postopera-tively,while the expression of HSP60 increases gradually and reaches the highest 48 h postoperatively.(3)The change of expression of Neuritin and HSP60 is closely related to the process of liver injury repair(P < 0.001), showing a certain change rule. They may have some biological effects through interactions,and participate in and promote the regeneration and repair of liver injury.

SUMMARY Thehumanembryonicstemcells(hESCs)serveasaself-renewable,genetically-healthy, pluripotent and single source of all body cells,tissues and organs.Therefore,it is considered as the good standard for all human stem cells by US,Europe and international authorities.In this study,the standard and healthy human mesenchymal progenitors,ligament tissues,cardiomyocytes,keratinocytes,primary neurons,fibroblasts,and salivary serous cells were differentiated from hESCs.The human cellular health-safety of NaF,retinoic acid,5-fluorouracil,dexamethasone,penicillin G,adriamycin,lead ace-tate PbAc,bisphenol A-biglycidyl methacrylate (Bis-GMA)were evaluated selectively on the standar-dized platforms of hESCs,hESCs-derived cardiomyocytes,keratinocytes,primary neurons,and fibro-blasts.The evaluations were compared with those on the currently most adopted cellular platforms.Parti-cularly,the sensitivity difference of PM2.5 toxicity on standardized and healthy hESCs derived fibroblasts, currently adopted immortalized human bronchial epithelial cells Beas-2B and human umbilical vein endo-thelial cells (HUVECs)were evaluated.The results showed that the standardized hESCs cellular plat-forms provided more sensitivity and accuracy for human cellular health-safety evaluation.

Objective To report a pedigree with X?linked dominant protoporphyria(XLDPP), and to detect 5?aminolevulinic acid synthetase 2(ALAS2)gene mutations in this pedigree. Methods A clinical investigation was performed in a pedigree with XLDPP, and relevant data were collected from family members. A next?generation sequencing method was applied to screen possible mutation sites, and Sanger sequencing was performed to determine pathogenic gene mutations. Dermoscopy was conducted to observe skin lesions in the patients with XLDPP, and the Fotofinder system and very high frequency (VHF) ultrasound system were utilized to assess the severity of photodamage. Liver and gallbladder ultrasonography as well as blood examination were performed for all the family members. Results A deletion mutation, c.1706?1709ΔAGTG, was detected in the ALAS2 gene on the X chromosomes of all the patients in this family, which led to replacement or loss of 19-20 C?terminal residues through transcriptional frameshifting, and eventually caused an increase in ALAS2 activity. In the patients with XLDPP, skin photodamage was relatively severe;protoporphyrin?induced hepatobiliary damage was observed and aggravated with age;anemia and iron deficiency occurred sometimes. Conclusion The deletion mutation c.1706?1709ΔAGTG of the ALAS2 gene may be the underlying cause of XLDPP in this pedigree.

Objective:To further improve the understanding of paroxysmal nocturnal hemoglobinuria (PNH), we retrospectively analyzed and summarized the clinical characteristics, treatment status, and survival status of patients with PNH in Zhejiang Province.Methods:This study included 289 patients with PNH who visited 20 hospitals in Zhejiang Province. Their clinical characteristics, comorbidity, laboratory test results, and medications were analyzed and summarized.Results:Among the 289 patients with PNH, 148 males and 141 females, with a median onset age of 45 (16-87) years and a peak onset age of 20-49 years (57.8% ). The median lactic dehydrogenase (LDH) level was 1 142 (604-1 925) U/L. Classified by type, 70.9% (166/234) were classical, 24.4% (57/234) were PNH/bone marrow failure (BMF), and 4.7% (11/234) were subclinical. The main clinical manifestations included fatigue or weakness (80.8%, 235/289), dizziness (73.4%, 212/289), darkened urine color (66.2%, 179/272), and jaundice (46.2%, 126/270). Common comorbidities were hemoglobinuria (58.7% ), renal dysfunction (17.6% ), and thrombosis (15.0% ). Moreover, 82.3% of the patients received glucocorticoid therapy, 70.9% required blood transfusion, 30.7% used immunosuppressive agents, 13.8% received anticoagulant therapy, and 6.3% received allogeneic hematopoietic stem cell transplantation. The 10-year overall survival (OS) rate was 84.4% (95% CI 78.0% -91.3% ) . Conclusion:Patients with PNH are more common in young and middle-aged people, with a similar incidence rate between men and women. Common clinical manifestations include fatigue, hemoglobinuria, jaundice, renal dysfunction, and recurrent thrombosis. The 10-year OS of this group is similar to reports from other centers in China.

Osteoarthritis is the most common degenerative cartilage disease. A large number of studies have shown the close association between epigenetics and osteoarthritis. Histone methylation is a type of epigenetic modification, and the link between histone methylation and osteoarthritis has also been revealed. In this article, we summarize the correlation between methylation levels of different histones and osteoarthritis in an attempt to explore the changes and regulation mechanisms of histone methylation in osteoarthritis. It has been shown that there are possible relations between the methylation levels of different amino acids on histone H3 and the pathological development of osteoarthritis; specifically, the rise of methylation level at the lysine 4 would aggravate the pathological development of osteoarthritis, while the the pattern of lysine 9 and 27 would be the opposite. These results indicate the possible existence of a complex network of histone methylation modifications. And the specific regulation of histone methylation levels in different positions may delay or prevent the occurrence and development of osteoarthritis.

Embryonic stem cells have unlimited proliferative capacity, which may provide a source of tendon stem/progenitor cells for tissue engineering. Experts of International Science and Technology Collaborative Program of Ministry of Science and Technology have developed a protocol consensus on differentiation of human embryonic stem cells into the tendon cells. The consensus recommends a protocol of two-step generation of human embryonic stem cells into tendon cells: the human embryonic stem cells are first differentiated into mesenchymal stem cells on different material surfaces; then with the scaffold-free tissue engineering tendon formed by high-density planting, the mesenchymal stem cells are induced into tendon cells under static or dynamic mechanical stimulation in vivo and in vitro. Tissue engineering tendon established in vitro by the protocol can be used as a model in toxicological analysis and safety evaluation of tendon-relevant small molecule compounds, medical materials and drugs.
Cell Differentiation ; Consensus ; Human Embryonic Stem Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Tendons ; cytology ; Tissue Engineering

ObjectiveBased on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry（UPLC-Q-Orbitrap-MS）， to identify the in vivo and in vitro chemical components of total phenolic acids in Gei Herba（TPAGH）， and to clarify the pharmacological effects and potential mechanisms of the effective part in promoting acute wound healing and inhibiting scar formation. MethodsUPLC-Q-Orbitrap-MS was used to identify the chemical components of TPAGH and ingredients absorbed in vivo after topical administration. A total of 120 ICR mice were randomly divided into the model group， recombinant human epidermal growth factor（rhEGF） group（4 mg·kg^-1）， and low， medium， and high dose groups of TPAGH（3.5， 7， 14 mg·kg^-1）， with 24 mice in each group. A full-thickness skin excision model was constructed， and each administration group was coated with the drug at the wound site， and the model group was treated with an equal volume of normal saline， the treatment was continued for 30 days， during which 8 mice from each group were sacrificed on days 6， 12， and 30. The healing of the wounds in the mice was observed， and histopathological changes in the skin tissues were dynamically observed by hematoxylin-eosin（HE）， Masson， and Sirius red staining， and enzyme-linked immunosorbent assay（ELISA） was used to dynamically measure the contents of interleukin-6（IL-6）， tumor necrosis factor-α（TNF-α）， vascular endothelial growth factor A（VEGFA）， matrix metalloproteinase（MMP）-3 and MMP-9 in skin tissues. Network pharmacology was used to predict the targets related to the promotion of acute wound healing and the inhibition of scar formation by TPAGH， and molecular docking of key components and targets was performed. Gene Ontology（GO） biological process analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis were carried out for the related targets， so as to construct a network diagram of herbal material-compound-target-pathway-pharmacological effect-disease for further exploring its potential mechanisms. ResultsA total of 146 compounds were identified in TPAGH， including 28 phenylpropanoids， 31 tannins， 23 triterpenes， 49 flavonoids， and 15 others， and 16 prototype components were found in the serum of mice. Pharmacodynamic results showed that， compared with the model group， the TPAGH groups showed a significant increase in relative wound healing rate and relative scar inhibition rate（P<0.05）， and the number of new capillaries， number of fibroblasts， number of new skin appendages， epidermal regeneration rate， collagen deposition ratio， and Ⅲ/Ⅰ collagen ratio in the tissue were significantly improved（P<0.05， 0.01）， the levels of IL-6， TNF-α， MMP-3 and MMP-9 in the skin tissues were reduced to different degrees， while the level of VEGFA was increased. Network pharmacology analysis screened 10 core targets， including tumor protein 53（TP53）， sarcoma receptor coactivator（SRC）， protein kinase B（Akt）1， signal transducer and activator of transcription 3（STAT3）， epidermal growth factor receptor（EGFR） and so on， participating in 75 signaling pathways such as advanced glycation end-products（AGE）-receptor for AGE（AGE/RAGE） signaling pathway， phosphatidylinositol 3-kinase（PI3K）/Akt signaling pathway， mitogen-activated protein kinase（MAPK） signaling pathway. Molecular docking confirmed that the key components genistein， geraniin， and casuariin had good binding ability to TP53， SRC， Akt1， STAT3 and EGFR. ConclusionThis study comprehensively reflects the chemical composition of TPAGH and the absorbed components after topical administration through UPLC-Q-Orbitrap-MS. TPAGH significantly regulates key indicators of skin healing and tissue reconstruction， thereby clarifying its role in promoting acute wound healing and inhibiting scar formation. By combining in vitro and in vivo component identification with network pharmacology， the study explores how key components may bind to targets such as TP53， Akt1 and EGFR， exerting therapeutic effects through related pathways such as immune inflammation and vascular regeneration.