Comparison studies of recombinant prourokinase(pro-UK) in various host cells,and expression vectors were carried out. Expression levels of vectors constructed in this study in different cell lines were compared. Mammalian cells expressing pro-UK were established. The levels of pro-UK expression in recombinant Namalwa, Vero and Sp2/0 cells are 200,12.5 and 50 IU/( 106 cells 24h). pro-UK purities separated by immunoaffinity chromatograph are above 90 %. Immunoabsorbent assay showed the ratio of pro-UK in CHO cells is lower than that from Vero and Namalwa cells. This study provide new host cells for pro-UK production.

The purpose of this study was to investigate how Zuogui pills from the Kidney-tonifying and Nourishing Yin formula, in combination with the gonadotrophin-releasing hormone antagonist cetrorelix, affected the ovarian local oxidative stress response in decreasing ovarian reserve (DOR) mice. All animal experiments were carried out in accordance with the guidelines and standards established by Jinan University's Experimental Animal Management Committee. Cyclophosphamide (CTX)-treated DOR mice were given Zuogui pills, cetrorelix, or a combination of the two drugs intragastrically. After treatment, there were changes in the estrous cycle, serum sex hormone levels, oxidative stress-related indexes, growth biochemical factor levels, and SIRT1/P53/P21 expression. In comparison to the model group, the Zuogui pills and the cetrorelix+Zuogui pills group had significantly prolonged estrous periods and shortened interestrous periods, and the cetrorelix+Zuogui pills group had a significantly shortened cycle length. Follicle-stimulating hormone (FSH) decreased and estradiol (E₂) increased in all treatment groups compared to the model group, oxidative stress indexes nitric oxide synthase (NOS), nitric oxide (NO), and reactive oxygen species (ROS) decreased, growth biochemical factors brain derived neurotrophic factor (BDNF) and growth differentiation factor 9 (GDF-9) concentrations increased significantly, and leukemia inhibitory factor (LIF) showed no significant change. SIRT1/P53/P21 immunohistochemical results revealed that, when compared to the model group, the expression of SIRT1 increased while the expression of P53 and P21 proteins decreased in all treatment groups, with the cetrorelix+Zuogui pills group having the largest decrease, with significant differences in all indicators. We conclude that cetrorelix combined with Zuogui pills for kidney nourishing and Yin recipe improved the oxidative stress response in the follicle by regulating the SIRT1/P53/P21 pathway, reducing peroxide product production, protecting ovarian function, and regulating ovarian hormone secretion, and its efficacy is superior to that of cetrorelix or Zuogui pills alone.

OBJECTIVETo investigate effect of connective tissue growth factors (CTGF) on secretion of extracellular matrix synthesis of meniscal fibrochondrocytes, expression of vascular endothelial growth factors (VEGF), and angiogenesis during the repair of meniscal tearing damage.

METHODSMeniscal fibrochondrocytes were isolated from the inner--1/2 of rabbits' meniscus by collagenase enzymatic digestion, centrifugal separation, and treated with 100 ng/ml CTGF in vitro. Characterization of fibrochondrocytes was identified by flow cytometry analysising CD31, CD44, CD45 and CD105, and was further tested by type II collagen immunocytochemistry. Changes in gene expression of meniscal fibrochondrocytes were monitored by quantitative real-time polymerase chain reaction. In vitro, the sections of the 3 mm of the longitudinal teared in the middle of the rabbit's meniscus, and then the defects were dealed with simple suture, suture and implanting with PBS-fibrin glue, sutured and implanting with 1.5 microg CTGF respectively. Expression and distribution of type I and II collagen and VEGF, the tearing healing were observed by fluorescence-immunohistochemisty analysis on the 1st week, the 4th week and the 10th week.

RESULTSQuantitative RT-PCR assays showed that type I and type II collagen,and VEGF mRNA expression in the 100 ng/ml CTGF group had been remarkably enhanced than in the PBS group on the 14th day. Consistent with these effects in vitro, fluorescence-immunohistochemical analysis revealed that in the group implanted with CTGF-fibrin glue, type I collagen, type I collagen and capillaries completely filled the defect on the 10th week postoperatively. In contrast, only soft tissue repair occurred after the PBS-fibrin glue was implanted.

CONCLUSIONCTGF can significantly promote extracellular matrix (I collagen, II collagen) of the meniscal avascular zone synthesis, and CTGF can greatly heighten the expression of VEGF activity at the same time in vitro, so that it can further enhance the repair of meniscal tearing damage in the avascular zone.

Animals ; Collagen Type I ; genetics ; Collagen Type II ; genetics ; Connective Tissue Growth Factor ; therapeutic use ; Gene Expression Regulation ; drug effects ; Male ; Menisci, Tibial ; surgery ; Rabbits ; Tibial Meniscus Injuries ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing ; drug effects

PURPOSETo discuss surgical technique, operative efficacy and clinical outcome of intramedullary fixation in the treatment of subtrochanteric femur fractures.

METHODSFrom February 2011 to February 2013, 76 cases of subtrochanteric femur fractures were treated by intramedullary fixation in our hospital, including 53 males and 23 females, with the age range of 37 -72 years (mean 53.5 years). According to Seinsheimer classification, there were 2 cases of type I, 7 type II, 15 type III, 23 type IV and 29 type V. Firstly, all patients underwent closed reduction with the guidance of C-arm fluoroscopy in a traction table. Two cases of type I and 3 cases of type III fractures had ideal closed reduction followed by internal fixation. The others needed additional limited open reduction. Radiographic examination was used to evaluate callus formation and fracture healing in postoperative 1, 3, 6 and 12 months follow-up. Functional recovery was evaluated by Harris Hip Scoring (HHS) system.

RESULTSPatients were followed up for 6-12 months. All fractures were healed except one patient with delayed union. The average bone union time was 4.5 months. According to HHS system, 65 cases were considered as excellent in functional recovery, 8 good, 2 fair and 1 poor. The proportion of the patients with excellent and good recovery was 96.05%.

CONCLUSIONIntramedullary fixation is feasible for the treatment of subtrochanteric femur fracture. The accuracy of intraoperative reduction and surgical skill are important for the clinical outcome and the patients' prognosis.

Adult ; Aged ; Female ; Femoral Fractures ; classification ; surgery ; Fluoroscopy ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged ; Prognosis ; Recovery of Function ; Treatment Outcome

OBJECTIVETo investigate the clinical effects of sialoendoscopically-assisted sialolithectomy for the sialolithiasis in the hilum of the submandibular glands.

METHODSBetween December 2005 and March 2011, treated 80 cases of radiography-verified sialiolithiasis in the hilum of the submandibular glands, The patients included 42 males and 38 females aged from 13 to 68 years. All these patients underwent sialoendoscopic observation and sialoendoscopically-assisted sialolithectomy and were followed up periodically for 3 - 6 months after operation. The success rate of stone removal, postoperative complications and clinical effects were analysed.

RESULTSThe stones were completely removed in 71 cases, and almost completely removed in 5 cases, with a successful rate of 95% (76/80). Among 76 successful cases, 8 were treated by basket entrapment, 59 by intraoral open surgery and 9 by both of these two techniques. Within 3 - 6 months' follow-up, 1 case suffered temporary lingual nerve parenthesis and two suffered ranula formation.

CONCLUSIONSSialoendoscopically-assisted sialolithectomy is a safe and effective gland-preservation technique for the patients with the hilum of the submandibular glands.

Adolescent ; Adult ; Aged ; Endoscopy ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Natural Orifice Endoscopic Surgery ; adverse effects ; Ranula ; etiology ; Salivary Duct Calculi ; surgery ; Salivary Ducts ; pathology ; Submandibular Gland ; surgery ; Young Adult

Objective To compare the histologic grade between biopsy and postoperative specimen in bladder urothelial carcinoma,and approach the state and the reasons of underestimate the histologic grade preoperative.Methods We retrospectively 82 cases of urothelial carcinoma at the Third Affiliated Hospi- tal of the Sun Yat-Sen University.For all the cases in this study,the histologic grade,using the 1998 World Health Organization and International Society of Urological Pathologists(WHO/ISUP)classification,was i- dentical when the biopsy specimen and postoperative specimen were compared.Results In this study,35 cases,28 cases and 19 cases were G_1、G_2、G_3 by biopsy preoperative,respectively;while 22 cases,32 cases、28 cases were G_1、G_2、G_3 postoperative,respectively.There were 24 cases(29.3%)underestimate the histo- logic grade by biopsy preoperative in the 82 cases,while 4 cases(4.9%)overestimate preoperative.The state of underestimate the histologic grade is correlated with the location of biopsy,tissue dose and the conser- vation of pathology judgment.Conclusions There were 24 cases(29.3%)underestimate the histologic grade by biopsy preoperative.We should pay more attention to this state of underestimate the histologic grade preoperative in the treatment of bladder urothelial carcinoma.

Objective:To investigate effect of connective tissue growth factors (CTGF) on secretion of extracellular matrix synthesis of meniscal fibrochondrocytes,expression of vascular endothelial growth factors (VEGF),and angiogenesis during the repair of meniscal tearing damage. Methods:Meniscal fibrochondrocytes were isolated from the inner-1/2 of rabbits'meniscus by collagenase enzymatic digestion,centrifugal separation,and treated with 100 ng/ml CTGF in vitro. Characteriza-tion of fibrochondrocytes was identified by flow cytometry analysising CD31 ,CD44,CD45 and CD105,and was further tested by typeⅡcollagen immunocytochemistry. Changes in gene expression of meniscal fibrochondrocytes were monitored by quanti-tative real-time polymerase chain reaction. In vitro ,the sections of the 3 mm of the longitudinal teared in the middle of the rab-bit's meniscus,and then the defects were dealed with simple suture,suture and implanting with PBS-fibrin glue,sutured and implanting with 1.5μg CTGF respectively. Expression and distribution of typeⅠandⅡcollagen and VEGF,the tearing healing were observed by fluorescence-immunohistochemisty analysis on the 1st week ,the 4th week and the 10th week. Results:Quantitative RT-PCR assays showed that typeⅠand typeⅡcollagen,and VEGF mRNA expression in the 100 ng/ml CTGF group had been remarkably enhanced than in the PBS group on the 14th day. Consistent with these effects in vitro ,fluores-cence-immunohistochemical analysis revealed that in the group implanted with CTGF-fibrin glue ,typeⅠcollagen,typeⅡcol-lagen and capillaries completely filled the defect on the 10th week postoperatively. In contrast ,only soft tissue repair occurred after the PBS-fibrin glue was implanted. Conclusion:CTGF can significantly promote extracellular matrix (Ⅰcollagen,Ⅱcol-lagen) of the meniscal avascular zone synthesis,and CTGF can greatly heighten the expression of VEGF activity at the same time in vitro,so that it can further enhance the repair of meniscal tearing damage in the avascular zone.

Objective To investigate the improvement effect and mechanism of Modified Kongsheng Zhenzhong Dan medicated serum on mitochondrial function of PC12 cells injured by oxygen-glucose deprivation/reperfusion(OGD/R)based on the SIRT1/PGC-1α pathway.Methods PC12 cells were used to construct OGD/R cell model in vitro.Cell grouping:normal group(10%FBS),model group(10%FBS),10%drug-containing serum group,5%drug-containing serum group(5%drug-containing serum+5%blank serum),10%blank serum group(control group).CCK-8 method was used to detect the cell survival rate,and the appropriate oxygen glucose deprivation time(2,4,6,8 hours)was screened.MTT assay was used to detect cell activity;mitochondrial stress test(MST)was used to detect the oxygen consumption rate(OCR).Apoptosis was detected by flow cytometry(Annexin V-PE/7-AAD double staining).The protein expression levels of SIRT1 and PGC-1α in PC12 cells were detected by Western Blot.Results Compared with the normal group,the activity of PC12 cells decreased significantly after oxygen-glucose deprivation for 2,4,6 and 8 hours(P＜0.05),and 6 hours of oxygen-glucose deprivation was selected as the time of subsequent experimental modeling.Compared with the normal group,the cell activity of the model group was significantly decreased(P＜0.05).The OCR values of basic respiration value,maximum respiration value,proton leakage,ATP production and standby respiration ability were significantly decreased(P＜0.05).The apoptosis rate was significantly increased(P＜0.05).The protein expression levels of SIRT1 and PGC-1α in cells were significantly decreased(P＜0.05).Compared with the model group,the cell activity of the Modified Kongsheng Zhenzhong Dan 10%and 5%drug-containing serum groups was significantly increased(P＜0.05).The OCR values of basal respiration value,maximum respiration value,proton leakage,ATP production and spare breathing ability were significantly increased(P＜0.05).The apoptosis rate was significantly decreased(P＜0.05).The protein expression levels of SIRT1 and PGC-1α in cells were significantly increased(P＜0.05).Conclusion The Modified Kongsheng Zhenzhong Dan medicated serum can improve mitochondrial dysfunction,inhibit neuronal apoptosis and promote neuronal survival in OGD/R-injured PC12 cells.The mechanism may be related to the activation of SIRT1/PGC-1α signaling pathway.

Background: Thymoma is an uncommon tumor without a widely accepted standard care to date. We aimed to investigate the clinicopathologic variables of patients with thymoma and identify possible predictors of survival and recurrence after initial resection. Methods: We retrospectively selected 307 patients with thymoma who underwent complete resection at the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College (Beijing, China) between January 2003 and December 2014. The associations of patients' clinical characteristics with prognosis were estimated using Cox regression and Kaplan–Meier survival analyses. Results: During follow-up (median, 86 months; range, 24–160 months), the 5- and 10-year disease-free survival (DFS) rates were 84.0% and 73.0%, respectively, and the 5- and 10-year overall survival (OS) rates were 91.0% and 74.0%, respectively. Masaoka stage (P < 0.001), World Health Organization (WHO) histological classification (P < 0.001), and postoperative radiotherapy after initial resection (P = 0.006) were associated with recurrence (52/307, 16.9%). Multivariate analysis revealed that, after initial resection, WHO histological classification and Masaoka stage were independent predictors of DFS and OS. The pleura (25/52, 48.0%) were the most common site of recurrence, and locoregional recurrence (41/52, 79.0%) was the most common recurrence pattern. The recurrence pattern was an independent predictor of post-recurrence survival. Patients with recurrent thymoma who underwent repeated resec-tion had increased post-recurrence survival rates compared with those who underwent therapies other than surgery (P = 0.017). Conclusions: Masaoka stage and WHO histological classification were independent prognostic factors of thymoma after initial complete resection. The recurrence pattern was an independent predictor of post-recurrence survival. Locoregional recurrence and repeated resection of the recurrent tumor were associated with favorable prognosis.

BACKGROUND: With the development of chemical synthesis technology, the application of anti-osteoporosis drugs incorporated into scaffolds to promote bone regeneration in osteoporotic bone defects has become a hot issue nowadays. OBJECTIVE: To introduce bone tissue engineering scaffolds, and to discuss and summarize the application of the scaffolds carrying anti-osteoporosis drugs for osteogenesis in osteoporotic bone defects. METHODS: PubMed, Web of Science, Springerlink, Medline, WanFang and CNKI databases were retrieved with "osteoporosis, bone defect, scaffold" as key words for relevant articles published from 2005 to 2017. Initially, 201 articles were retrieved, and finally 64 articles were selected for further analysis. RESULTS AND CONCLUSION: With the development of biochemical synthesis technology, various types of tissue-engineered bone scaffolds have been used for the treatment of osteoporotic bone defects. For example, the modified calcium phosphate cement, β-tricalcium phosphate, and hydroxyapatite all can promote bone regeneration. However, the ability of conventional scaffolds to promote bone regeneration is severely damaged under the state of osteoporosis. In this case, an anti-osteoporosis therapy becomes necessary. Considering severe adverse effects of systemic anti-osteoporosis drugs and limited local bone regeneration, the combination of scaffold implantation and local drug delivery can promote osteoporotic bone defect healing. Importantly, to incorporate bone regeneration inducers into scaffolds and steadily release with their biological activity is the key to success. Bisphosphonate, strontium, bone morphogenetic protein 2 and parathyroid hormone, traditional Chinese medicine and mesenchymal stem cells are the drugs commonly combined with tissue-engineered bone scaffolds. With the development of biotechnology, genetically modified stem cell-scaffold composites have gradually appealed to researchers.