Humans ; Perchlorates ; toxicity ; Thyroid Gland ; drug effects

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

Liver Cirrhosis ; Metallothionein

Objective To investigate the effect of leptin on endoplasmic reticulum stress related protein after focal cerebral ischemia in rats. Methods Ischemia was induced by occluding the middle cerebral artery in rats brain using the filament occlusion method. Forty SD rats were randomly divided into sham operation group, cerebral is-chemia group and leptin-preconditioning group. Leptin was injected subcutaneously before occlusion of blood vessel. Longa 5 score neurological function scale, body weight and brain edema changes were measured 6 hours after MCAO, and the brain was removed to detect the endoplasmic reticulum marker protein: glucose-regulated protein 78 (GRP78) and C/EBP-homologous protein (CHOP) by immunohistochemical method. Results There was no difference in the body weight changes between leptin-preconditioning group and ischemic group. In the leptin-preconditioning group, the neurological function score (1.90±0.31 vs. 2.50±0.52, P<0.05) and the degree of brain edema (3.60±0.52 vs. 7.70±0.94, P<0.001) were significantly lower than those in the cerebral ischemia group. Moreover, the expression of GRP78 in leptin-preconditioning group was significantly higher than that in ischemia group (48.69 ±5.06 vs. 35.78± 4.35, P<0.01), and the expression of CHOP was significantly lower than that of ischemia group (60.24 ±4.11 vs. 38.81±5.34, P<0.01). Conclusion Leptin can reduce the neurological deficit and may be associated with the up-reg-ulation of GRP78 protein, and down regulation of CHOP protein to weaken the endoplasmic reticulum stress caused by cerebral ischemia

Objective To investigate the survival of advanced stage periampullary carcinoma and its prognostic factors.Methods The clinical data and follow up of 134 patients with advanced stage periampullary carcinoma admitted to our hospital between January 2007 and December 2010 were retrospectively collected and analyzed.Results Of 134 patients,there were 83 males and 51 females.The mean age±S.D.was 62.49±11.41 (28-83).For the 86 patients who underwent surgery,32 had extensive peritoneal metastases on surgical exploration,27 had metastases/involvement of the liver or stomach,and 27 had major vessel infiltration.For these 86 patients,55 received surgical exploration,whereas 31 had surgical exploration plus bypass anastomosis. 45 patients received chemotherapy,while 67 received chemotherapy plus radiotherapy.The improvements in quality of life and in clinical response rate after concurrent chemoradiotherapy or chemotherapy alone were 73.1％ and 57.8％,respectively.The overall survival was 14.27±1.06 months.The median survival time was 11 (8.95±13.05) months.The 1- and 3-year survival rates were 43％ and 10％,respectively.Karnofsky performance status,and the presence or absence of jaundice co-related with poor prognosis on single factor analysis.The Karnofsky performance status was an independent survival predictor on multifactor analysis.Conclusions The prognosis of patients with advanced stage periampullary carcinoma was poor.Chemotherapy with or without radiotherapy had similar impact on overall survival,Karnofsky performance status was an independent survival predictor.

We developed and designed a new type of artificial trachea. The basic structure of the artificial trachea was polytetrafluoroethylene vascular prosthesis linked with titanium rings on both sides. Dualmesh was sutured on titanium rings. This experimentation follows the replacement of trachea in dogs with a combined artificial trachea to investigate the feasibility of this type of prosthesis. Sixteen dogs were implanted with the combined artificial trachea after resection of 5 cm of cervical trachea. The 5 cm-long trachea of dogs on the necks were resected and the reconstruction of the defect of the trachea was performed with trachea prosthesis. According to the method of trachea reconstruction, the models were divided into 2 groups, artificial trachea implantation group (the control group, n = 8) and group of artificial trachea implantation with growth factor (the experimental group, n = 8). Then computer tomography scan (CT), bronchoscope and pathologic examination were conducted periodically to observe the healing state of the hybrid artificial trachea. None of the dogs died during operation of cervical segmental trachea construction. But four dogs in the control group died of apnea in succession because artificial trachea was displaced and the lumen was obstructed, while 2 dogs died in the experimental group. In the first month there was granulation around anastomosis with slight stenosis. The rest of dogs were well alive until they were sacrificed 14 months later. The mean survival time of the experimental group was longer than that of the control group. The rate of infection, anastomotic dehiscence, severe stenosis and accidental death in the experimental group were lower than the control group (P < 0.05). Artificial trachea was encapsulated by fibrous tissue and no mucous membrane was seen in the lumen of the artificial trachea. The artificial trachea can be used to reconstruction of the defect of the trachea with long-term survival of the animals. The unique design of artificial trachea reduces stenosis around anastomosis effectively but infections and split or displacement of the artificial trachea are still major problems affecting long-term survival of the animals. Application of growth factors to a certain extent promotes tissue healing by changing the local environment.
Animals ; Artificial Organs ; Dogs ; Prostheses and Implants ; Prosthesis Design ; Prosthesis Implantation ; Reconstructive Surgical Procedures ; Titanium ; Trachea ; surgery

This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.
Aconitine ; analogs & derivatives ; chemistry ; pharmacokinetics ; Animals ; Biological Availability ; Dogs ; Drug Stability ; Female ; Male

Objective To prepare a novel radiolabeled FR?positive tumor targeting agent 99 Tcm?( HYNIC?NHHN?FA) ( EDDA) and evaluate its biological properties. Methods FA derivative FA?NHHN?HYNIC was synthesized and radiolabeled with 99 Tcm using EDDA as a coligand. The radiochemical purity, octanal/water partition coefficient and in vitro stabilities of the complex were studied after purified by HPLC. In vitro cellular uptakes were performed on FR?positive KB cells ( human oral epidermoid carcinoma cells) . Biodistribution and microSPECT/CT imaging were investigated on normal Kunming mice and nude mice bearing KB tumors, respectively. Results The radiochemical purity of the complex was over 95% after pu?rified by HPLC. It displayed high stability both in saline and in serum. It also exhibited high specific FR binding in FR?positive KB cells in vitro. The binding ratio was (6.76±0.60)%1 h after incubation, and de?creased to (0.24±0.02)% after adding excessive FA. The results of biodistribution showed high kidney up?take in normal mice, and the uptake reached (21.79±9.79) %ID/g 0.5 h after injection. Flank KB tumors were clearly visualized with 99 Tcm?( HYNIC?NHHN?FA ) ( EDDA ) by microSPECT/CT imaging at 2 h postinjection, and the uptake could be inhibited by excessive FA. Conclusions 99 Tcm?( HYNIC?NHHN?FA) ( EDDA) exhibits good pharmacokinetic properties, suggesting its potential as a promising FA targeting agent for tumor imaging.

Objective To observe the pathological changes of bone hydatid cyst of meriones meridianus after radiation therapy,and to investigate the clinical effect of radiotherapy on bone hydatid disease.Methods Ascus was dissected sterilely from sheep liver that naturally infected with Echinococcus granulomas,sheared and sac skin removed.Then it was washed and precipitated with 0.9％ sterile saline for 3 times,and scolex was HE stained and counted,from which a 20 ml suspension was made containing 12 × 106/L of scolex.Health meriones meridianus (referred to as gerbil) 140,male and female were in each half,aged 2 to 3 months,body weight(38 ± 6)g,were involved in the study.Gerbil was injected a 0.2 ml suspension containing Echinococcus granulomas scolex into hind tibial periosteum,and X-ray was taken 12 months after the injection.According to the bone destruction in the vaccination site,gerbil hindleg tibia with clear jagged bone destruction was treated as inclusion criteria,and 72 animal were selected as gerbil bone hydatid disease animal models,male and female were in each half.A tatal of 72 gerbils were randomly divided into 4 groups:control group,40 beequerel(Gy) group,50 Gy group and 60 Gy group,18 rats in each group,male and female in each half.The model animals were treated with radiotherapy for 5 times,with 2 d interval,and radiation dose was 300 cGy/min.Each group of gerbils was sacrificed after radiotherapy,bone Echinococcus granulomas cysts was taken out sterilely,and observed by light and electron microscope.Intracapsular cyst fluid was extracted,washed and precipitated with 0.9％ sterile saline repeatedly,and and the pellet was HE stained for observation of scolex morphology and activity by light microscope.Results The morphology and activity ofEchinococcus granulomas in cystic fluid in control group were normal; the morphology and activity of Echinococcus granulomas were still normal in the 40 Gy group,and Echinococcus granulomas was not stained red; but those were abnormal,deformation and atrophy and stained red in the 50 Gy group; and were stained red,deformed,fractured and were wrapped by unknown in the 60 Gy group.By light microscope,the germinal layer,cuticle layer,brood capsule and histological structure of protoscolex were basically normal in irradiated region in the control group.The pathological changes of hydatid cyst in the 40 Gy group were mainly degeneration,structure of hydatid cyst was abnormal,stratum corneum was extensive edema,germinal layer became thinner and the fertile cyst was rare.The main pathological change of hydatid cyst in the 50 Gy group was that corneous layer was widely fractured,and the germinal layer was edema,buckling folds,cells decreased,rare seen brood capsule and scolex; the main pathological changes of hydatid cyst were mainly necrosis in the 60 Gy group,cuticle was extensive fault,stratum corneum and germinal layer was separated,germinal layer was atrophy and disorder,no brood capsule and scolex.By electron microscope,cuticle structure of Echinococcus granulomas cyst was clear,microvillus arranged neatly,morphology and structure of the cell and organelle in cytoplasm were normal in the control group.There were many inflammatory cells infiltrating germinal layer of Echinococcus granulomas cyst,microfilament and contents in microfilament were reduced in the 40 Gy group.Microvillus of Echinococcus granulomas disappeared,nuclear membrane was unclear,endoplasmic and mitochon eclasis,lymphocyte nuclear chromatin was clumping and edge set and in circular permutation in the 50 Gy group.Microvillus disappeared,perinuclear membrane indistinct and ruptured,parts of nucleoli were fragmented and marinated,endoplasmic reticulum was extensive expansion,mitochondria was pyknosis and obvious vacuolization,lymphocyte nuclear chromatin clumping and edge set,lysosomes and macrophage emerge in the 60 Gy group.Conclusions Radiotherapy can destroy the morphology and structure of bone hydatid cyst,radioactivity at 50 Gy has a lethal effect on hydatid cyst.Radiation treatment of bone hydatid disease has a good clinical effect.

Objective: To explore the blood-saving effect of autologous platelet-rich plasma (PRP) back-transfusion in patients with Stanford type A aortic dissection surgery. Methods: A total of 59 consecutive patients who received Stanford type A aortic dissection surgery in our hospital from 2013-01 to 2015-10 were studied. The patients were at the age of (50±6) years with mean body weighting at (80±12) kg and were randomly divided into 2 groups: Traditional (T) group,n=31 and Autologous PRP back-transfusion (P) group,n=28. Blood levels of Hb, platelet counts, PT, APTT were measured at pre-induction of anesthesia (T1), before CPB (T2), prior ifnishing of CPB (T3) and at 1 h (T4), 24 h (T5), 48 h (T6) after the operation. The in-operative, 48 h post-operative volumes of allogeneic blood transfusion and the volume of chest tube drainage at 48h after operation were recorded; the complication occurrence at peri-operative period was recorded. Results: In P group, whole blood processing volume was (1269±197) ml, PRP volume was (753±78) ml, PRP separation time was (35±9) min and the separated platelets were about (22±3)% of total platelet counts. Compared with T group, P group had decreased Hb at T2 (131.0±15.0) g/L vs (101.0±10.0) g/L, decreased platelet counts at T3 (115.0±51.0)×109 /L vs (83.0±23.0)×109/L, while increased platelet counts at T4 (103.0±25.0)×109/L vs (151.0±27.0)×109/L, T5 (105.0±25.0)×109 /L vs (147.0±39.0)×109/L and T6 (101.0±26.0) ×109/L vs (149.0±35.0)×109/L, allP<0.05; P group presented reduced PT at T4 (17.6±2.1) s vs (14.1±1.1) s and T5 (17.6±2.7) s vs (13.5±0.8) s, allP<0.05. The in-operative transfusions of platelet, plasma, cold precipitation and post-operative transfusions of red blood cells, platelets, plasma, cold precipitation and the volume of chest tube drainage at 48h after operation were less in P group,P<0.05. Compared with T group, P group had the lower rates of acute post-operative lung injury (32.1% vs 19.4%), shorter mechanical ventilation time (69.1±5.9) h vs (43.1±1.5) h and ICU staying time (8.1±2.8) d vs (5.3±1.1) d, allP<0.05. Conclusion: Autologous PRP back-transfusion could reduce the post-operative bleeding and allogeneic blood transfusion for Stanford A aortic dissection surgery, it has obvious blood-saving effect.