Objective: To establish a determination for Ricinus communis L.and Carthamus tinctorius L.could in Anchan Emulsion. Methods: The determinations were carried out by GC. Chromatographic conditions were: using Chromosorb DEGS as a stationary phase column temperature 180 ?C , flame ionization detecter.Rusults:The content limit of Linoleic acid wasn't lower than 20%, The content limit of Ricinoleic acid wasn't lower than 20%.Conclusion: The established methods is simple, feasibl and reproducible. This study provides a method for the quality control of Anchan Emulsion.

BACKGROUND:Transformation growth factor beta 1 is mostly used to induce the chondrogenic differentiation of bone marrow mesenchymal stem cel s, but there is a poor induction efficacy.
OBJECTIVE:To explore the chondrogenic differentiation of bone marrow mesenchymal stem cel s co-cultured with articular chondrocytes or induced by transforming growth factor beta 1.
METHODS:Articular chondrocytes and bone marrow mesenchymal stem cel s from SD rats were harvested and divided into 1:2, 2:1, 1:1 concentration groups. Cel s induced by transforming growth factor beta 1 acted as control group. After 20 days of induced culture, MTT was used to detect cel viability, alcian blue colorimetric assay was applied to measure glycosaminoglycan content, and western blot assay was employed to determine the expression of col agen type II.
RESULTS AND CONCLUSION:The absorbance value in the control group was significantly lower than that in the 1:1 and 2:1 groups (P<0.05). Glycosaminoglycan content and protein expression of col agen type II were also lower in the control group than the 1:2, 1:1, 2:1 groups. But there was no difference between 1:1 and 2:1 groups (P>0.05). The results show that bone marrow mesenchymal stem cel s co-cultured with articular chondrocytes can be induced to differentiate into chondrocytes, and meanwhile, there is a saturation phenomenon during the chondrogenic differentiation of bone marrow mesenchymal stem cel s.

Objective To investigate the apoptotic effect of the transmembrane form vaccine of human blood group A mimotope on malignant melanoma cell line B16. Methods B16 cells were transfected with different recombinant plasmid through Lipofectamine 2000 and incubated with different concentration of monoclonal anti-A antibody at 2.5 μg/ml, 5 μg/ml,10 μg/ml and 20 μg/ml. Apoptosis rate of cells was determined with Annexin Ⅴ/PI double staining by flow cytometry. Results Apoptosis rate to P/F-M-pIRES group B16 cells was 74.74% when anti-A monoclonal antibody concentration was 10 μg/ml; apoptosis rate of plasmids carrying peptide/Fas fusion gene such as P/F-M-pIRES group and P/F-pIRES group were significantly higher than M-pIRES group and pIRES group. The apoptosis rate was statistically significantly different between different recombinated plasmid groups (F＝669.707,P＜0.01). The apoptosis rate was statistically significantly different between different antibody groups (F＝106.596,P＜0.01). The interaction between recombinated plasmid groups and antibody groups was statistically significant (F＝34.806,P＜0.01). Conclusions The transmembrane form vaccine of human blood group A mimotope could induce B16 cell apoptosis in vitro. This vaccine may be a promising candidate for potential malignant melanoma therapy.

OBJECTIVE: To study the modulation of Jianjining Recipe (JJNR), a traditional Chinese compound herbal medicine for invigorating spleen and kidney on differential protein expression in spleen of rats with experimental autoimmune myasthenia gravis (EAMG). METHODS: EAMG rats were randomly divided into four groups: untreated group, JJNR-treated group, Qiangji Jianli capsule (QJJLC, a traditional Chinese compound herbal medicine)-treated group and prednisolone acetate (PA)-treated group. After therapeutic intervention with the above drugs for four consecutive weeks, the level of differential protein expression was analyzed by two-dimensional electrophoresis and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Twelve differential proteins were identified by comparing EAMG rats and normal rats. The levels of allograft inflammatory factor-1, peroxiredoxin I and actin-related protein 2/3 complex subunit 5 were significantly regulated by JJNR (P<0.01). These proteins were closely associated with immune response and cell movement. CONCLUSION: The results suggest that there are differential protein expressions between EAMG rats and normal rats. Furthermore, as a Chinese medicine prescription with effect of invigorating spleen and kidney, JJNR can effectively regulate the levels of some EAMG-related protein expression.

Background The retinal degeneration 11 (rd11) mouse is a newly discovered naturally occurring recessive animal model with lysophosphatidylcholine acyltransferase 1 (Lpcatl) mutation.Previous studies showed that the photoreceptor cells are characterized by typical rod-cone degeneration pattern in rd1 1 mice,while cone degeneration pattern in rd11 mice is unclcar.Objective Using immunofluorescence staining techniques with retinal wholemount,we aim to clarify the degeneration patterns of cone-function related M-opsin or S-opsin in different ages of rd1 1 mice.Methods A total of thirty rd1 1 and C57BL/6J mice at postnatal (P) day 14,28,42 (five in each age group) were sacrificed and retinal wholemounts were prepared.Immunohistochemistry was performed to identify the expression of M-opsin or S-opsin in retinal wholemounts,which were photographed with a fluorescent microscope.Cone opsins were compared between rd1 1 retinas and age-matched normal C57BL/6J retinas by manually counting the opsin positive cone cells in different quadrants of the retinas.Results The number of M-opsin or S-opsin positive fluorescent dots in each quadrant was similar at all ages of normal C57BL/6J retina.M-opsin positive fluorescent dots in dorsal/temporal,ventral/temporal,dorsal/nasal and ventral/nasal quadrants of rdl 1 retina at P28 were (414±32),(300± 8),(324 ± 22) and (250± 20)/0.037 mm2,which were lower than the age-matched normal C57BL/6J mice (t =4.114,15.225,7.505,17.990,all at P＜0.05).At the same time the S-opsin positive fluorescent dots in P28 rd11 were (8 ±4),(175 ± 16),(74 ± 13) and (315 ±20)/0.037 mm2,with significant decrease in comparison with those in the age-matched normal C57BL/6J mice (t =8.555,17.076,21.637,13.498,all at P＜0.05).With the development of retinal degeneration in rd11 mice,the M-opsin degeneration spread from central to ventral,nasal and then to temporal and dorsal peripheral retina;and the S-opsin loss started from dorsal/temporal to ventral/nasal retina.Conclusions Most of the M-opsin and S-opsins,especially the S-opsins in rd11 mice,degenerate in 6 weeks.Retinal wholemount and cone opsin immunofluorescent staining provide a useful tool to show the cone degeneration pattern and to evaluate the therapeutic efficiency in ongoing gene therapy study.

Background Trans-corneally subretinal injection in rodent model is a useful method for genetic therapy,stem cell transplantation and the study on the ophthalmic research.Standarized operation process is critical for the successful treatment.However,there is no literature to report the detailed procedure and the influence of this technique on morphology and function of retina.Objective This sudy was to introduce a method of trans-corneally subretinal injection and evaluate its influence on the morphology and function of retina.Methods Trans-corneallly subretinal injection was performed on the left eyes of 2-month-old SPF C57BL/6J mice after dilation of pupils.A 301/2G disposable needle was used to puncture the cornea within the pupil area near limbus and avoid touching the lens and irises under eye surgery microscope.Then,a 33G blunt needle was used to insert into the vitreous and toward subretinal space via corneal puncture.Normal saline with 0.1％ fluorescein sodium of 1 μl was slowly injected into the space,and 2.5％ hydroxypropyl methylcellulose was dropped on ocular surface for the observation of the fundus clearly.According to the percentage of the retina filled with subretinally injected solution,the experimental eyes were divided into 80％-100％ area group,50％-70％ area group after injection,and the mice in the pseudo-injected group,in which injection procedure stopped just before the solution was pushed in to the subretinal space did not inject any solution after punctured.The right uninjected eyes of the mice served as normal control group.Four eyes were selected for each group.The structural changes were evaluated by optical coherance tomograpby (OCT) 1 day,2 days,3 days and 5 weeks after injection,and retinal function was assessed by the recored of electroretinography (ERG) 5 weeks after injection.The retinal sepcimens were prepared to examin the morphological changes by hematoxylin and esosin staning.The use of care followd the Regulations for the Administration of Affair Concerning Experimental Animals of Zhejiang Province.Results About 70％ of the injected eyes showed that retinal blebs filled with injected green fluorescein solution occupied 50％ or more retinal area with minimal damages.The focal detachment between neurosensory retinal layer and retinal pigment epithelium (RPE) was exhibited 1 day postinjection,and almost all the retinas retached 2 days after injection.In the fifth week after injection,the amplitudes of ERG b wave were (386.25±37.88),(357.50±41.03),(324.25±53.45) and (410.50±14.88) μV in the sham operation group,50％-70％ area group,80％-100％ area group and normal control group,respectively,showing a significant difference among the 4 groups (F=3.574,P=0.047),and the amplitudes of b wave in the normal control group were higher than those in the 80％-100％ area group (all at P ＜ 0.05).The detachment between retinal neuroepithelium layer and RPE layer,cell proliferation and transposition in the outer nuclear layer were dispalyed under the light microscope in the sham operation group,50％-70％ area group and 80％-100％ area group,and the disordered outer segment of photoreceptors at the injecting area was seen in the 50％-70％ area and 80％-100％ area groups at five weeks after injection.However,retinal sructure and morphology were normal at the non-injection area.Conclusions Trans-corneally subretinal injection is an effective and safe way for subretinal injection.

Objective To evaluate clinical features, therapeutic regimen and prognosis of unexplained rhabdomyolysis. Method Clinical manifestations, therapeutic regimen and prognosis were recorded in 23 patients,who were admitted to The First Affiliated Hospital of Nanjing Medical University 13 to 27 August,2010.The 23 patients were diagnozed as unexplained rhabdomyolysis. Results The patients all presented myalgia of upper body,like neck,waist and back,maybe with asthenia, nausea,dyspnea,abdominal pain, red urine or changed color of urine. Laboratory examination: obviously step-up of creatine kinase [CK: (4655 ± 2556) U/L( normal: 25 ～ 190U/L) AST:(141 ±78) U/L(normal:10～45 U/L),LDH:(348± 127) U/L(normal: 110～ 250 U/L)]and myoglobin[( Mb ＞ 1000 μg/L (normal: 0 ～ 50 μg/L)]. Therapeutic regimen included treatment of the underlying diseases, volume repletion, alkalization and dealing with the complications. No patients developed acute renal failure.All the patients recovered. Conclusions Rhabdomyolysis is a syndrome with different clinical manifestations.However, early diagnosis, proper treatment could prevent serious complications,and prognosis is good.

Stargardt disease (STGD) is an inherited disorder of retinal pigment epithelium.Three genes have been found to be implicated in STGD including Abca4 (adenosine triphosphate-binding cassette,sub-family A,member 4),Elovl4 (elongation of very long chain fatty acids protein 4) and Prom1 (prominin-1).Target genes can be delivered to the retina by various methods such as lentivirus (LV) vectors,adeno-associated virus (AAV) vectors and non-viral nano-particles.The Abca4-/-,Elovl4-/-and Prom1-/ mice model are used to study the pathogenesis mechanism and treatment of STGD.Retinal function improved significantly upon gene therapy in these models.Based on these works using animal model,phase Ⅰ /Ⅱ a clinical trial of Abca4-associated STGD gene therapy are underway.As a LV vector,equine infectious anemia virus (EIAV) is used to carry the Abca4 gene.These studies will evaluate three dose levels of the EIAV vector for safety,tolerability and biological activity.Moreover,some preclinical attempts to deliver Abca4 via AAV have been made using a modified AAV vectors because of the large size of the ABCA4 cDNA.The good responses in animal models render STGD a very attractive object for human gene therapy after the successful of the phase Ⅰ /Ⅱ clinical trials of Leber's congenital amaurosis.

Objective To investigate the effects of preemptive analgesia with parecoxib sodium on postoperative analgesia and delirium after nerve injury-free surgery for fracture of thoracic and lumbar vertebrae and to promote the postoperative rehabilitation of the patients. Method 80 patients meeting the criteria were selected. and randomly divided into observation group and control group.40 patients each group. The observation group used parecoxib sodium for preemptive analgesia. while the control group used sufentanil. and the analgesia effects and the incidences of delirium were observed. Results The differences in operative time and intra-operative blood loss between the patients of the two groups were statistically insignificant. In 2 h. 6 h. 12 h.24 h and 48 h after the surgery.the VAS score and the accumulative time of intravenous self-controlled analgesia pump being pressed of the observation group were significantly lower than those of the control group. and the differences were statistically significant (P<0.05). The first time for the patients of the observation group to press the intravenous self-controlled analgesia pump is (3.84±0.62) h after the surgery, is significantly later than that of the control group (1.05±0.47)h.and the difference is statistically significant (P<0.05). The incidence of delirium in 7 days after the surgery in the patients of the observation group was 10.00%. and is significantly lower than that of the control group (25.00%) (P<0.05). Conclusion Using parecoxib sodium for preemptive analgesia before nerve injury-free surgery for fracture of thoracic and lumbar vertebrae can elevate the postoperative analgesia effects of the patients.decrease the incidence of postoperative delirium, and is highly safe and consequently worthy of clinical application.

Objective To assess the clinical value of vascular enhancement technology (VET) in diagnosis of fetal pulmonary sequestration (PS).Methods Color Doppler velocity (CDV),color Doppler energy (CDE) and VET were performed in 11 fetus with PS to display the vessels in region of interest.Results The abnormal vessels in region of interest were displayed in distinct degree with CDV,CDE and VET.The courser and disposition of the great vessels were displayed with CDV and CDE,while lumen of blood vessel was clearly discriminated with VET,while CDV depicted the direction of blood flow.Satisfaction assessment of the VET image was superior than those of CDV and CDE image (P<0.05).No difference was found between the satisfaction assessment of the CDV and CDE images (P>0.05).Conclusion VET can improve the contrast resolution,having superiority in displaying vessels and fine branches,therefore has clinical value in the diagnosis of fetal PS.