Objective:To transfect human osteoprotegerin( o pg) gene into Cos-7 cells.Methods:The primers were designed b ased on the human opg cDNA sequence,total mRNA was isolated from 293 cells and RT-PCR was performed.The fragment of opg cDNA was inserted into pSecTag 2/B vector and sequenced by automatic sequence analyzer. The recombined plasmid was transfected into Cos-7 cells by Lipofectamine 2000 and OPG protein expressi on in Cos-7 cells was determined by immunohistochemistry and Western blot. Results:The sequence of opg cDNA from 293 cells obtained by RT- PCR was identical to the sequence provided by GenBank [gi:33878056]. The fragm ents of the recombinant plasmid digested with Hind Ⅲ,EcoR I and BamH Ⅰ and exa mined by 10 g/L agarose electrophoresis were consistent with predicted size.OPG over-expressing Cos-7 cells was selected and confirmed by immunohistochemistry and Western blot.Conclusion:Human opg gene may be transfect ed into Cos-7 cells.

Objective: To break the seed dormancy and improve the germination rate of the Nirtaria sibirica seeds. Methods: N. sibirica seeds were treated with soaking in warm water, storing with moisture sand, and immersing seed in H2SO4 (98%) and gibberellic acid (GA), then the germination rate, germination vigor, and germination peak period of the seeds were determined. Results: The germination rate and germination vigor were 30% and 10% under soaking in warm water. The seeds began to germinate on the day 7 and reached the germination peak on the day 10.The germination rate and germination vigor were 55.5% and 26.5% under storing with moisture sand for 37 d. The seeds began to germinate on the day 2 and reached the germination peak on the day 10.The germination rate and germination vigor were 86.8% and 60.3% under storing with moisture sand for 95 d. The seeds began to germinate on the day 2 and reached the germination peak on the day 7.The germination rate and germination vigor were both 90.0% under being treated with 98% H2SO4 for 2 h. The seeds began to germinate on the day 1 and reached the germination peak on the day 4.Seeds after being treated with 98% H2SO4 for 2 h were sowed on the dark medium of MS + 0.5 g/mL BA + 0.5 g/mL GA, and the germination rate and germination vigor both reached to 98.1%. The seeds began to germinate on the day 1 and reached the germination peak on the day 4. Conclusion: The dormancy of N. sibirica seeds is caused by hard seed vessels. The best way of breaking the seed dormancy is first treated with 98% H2SO4 for 2 h and then cultured in the dark medium of MS + 0.5 g/mL BA + 0.5 g/mL GA, which could effectively break the dormancy of hard seeds and reach a high germination rate.

Objective To investigate the changes of interleukin-17A(IL-17A)and interleukin-22(IL-22)in the lung of cigarette smoke exposed mice and the effect of smoking cessation and N -acetylcysteine (NAC )treatment . Methods BALB /c mice in experimental groups were exposed to cigarette smoke.Then the smoke-exposure was stopped and mice were treated with NAC gavage.The mice were executed 1,2,and 3 months after smoking cessa-tion.Lung tissue sample and bronchoalveolar lavage fluid(BALF)were collected.HE staining was used to observe the pathologic changes of the lung and ELISA was used to measure the level of IL-17A and IL-22.Results Con-siderable emphysematous changes was found in the lung of mice exposed to cigarette smoke.Compared with the controls,the level of IL-17A and IL-22 elevated remarkably in pulmonary tissue and BALF after smoking exposure and declined gradually after smoking cessation.Additional NAC gavage treatment enhanced the decline tendency. Conclusion IL-17A and IL-22 might play a complex role in the chronic inflammatory changes of lung in mice ex-posed to cigarette smoke.

Objective To explore the deficiency of upper limb movement in children with autism and its related factors. Methods 30 au-tism children aged 7-13 years old, and 30 healthy children aged 7-8 years from February to May, 2014 were enrolled. Their range of motion, comprehensive strength and cooperation, endurance, and muscular tension were tested. Results All the indexes of autism children lagged be-hind the healthy children (P<0.01), expect the muscular tension of the right upper limb (P>0.05). The endurance of boys was better than girls (P=0.020), and there was no difference in other aspects between them (P>0.05). The type of autism for more upper limb movement cor-related with obstruction of Jianjing acupoint (GB21) (r=0.515, P=0.013), and was significantly different with the type of less limb move-ment (χ2=8.533, P=0.003). Conclusion Compared with healthy children, the joint motion is relatively flexible, but lack of strength, and with poor persistence and unbalance development between left and right upper limbs. Motor rehabilitation should be conducted according to their movement features and the condition of meridian.

OBJECTIVE:To evaluate the bioequiavailability of the domestic and the imported paracetamol and oxycodone tablets.METHODS:The blood concentrations of paracetamol and oxycodone in22healthy male volunteers were determined by HPLC-MS after a single dose orally1tablet of domestic or imported oxycodone tablet by a randomized crossover way.RE?SULTS:The main pharmacokinetic parameters of the domestic and the imported oxycodone tablets were as follows:C max were(10.4?2.2),(11.1?3.3)?g/L,respectively;t max were(1.05?0.35),(0.92?0.40)h,respectively;t 1/2 ke were(5.36?0.91),(5.53?1.25)h,respectively;AUC 0～t were(44.2?7.9),(44.5?8.3)(?g?h)/L,respectively;AUC 0～∞ were(49.3?9.4),(51.0?11.6)(?g?h)/L,respectively;the relative bioavailability of the domestic preparation was(102.8?27.4)%.The main pharmacokinetic parameters of the domestic and the imported paracetamol were the following:C max were(4612?696),(4592?825)?g/L,respectively;t max were(0.94?0.28),(0.96?0.23)h,respectively;t 1/2 ke were(3.99?0.77),(4.05?0.83)h,re?spectively;AUC 0～t were(15732?3450),(16265?3858)(?g?h)/L,respectively;AUC 0～∞ were(16618?3545),(17205?4194)(?g?h)/L,respectively;the relative bioavailability of the domestic one was(97.6?10.3)%.CONCLUSIONS:The2preparations are bioequivalent.

Objective To explore the effect of glial cells on cerebral neuron damage induced by amyloid beta protein (Aβ) in Alzheimer′s disease rat .Methods 20 male Wistar rats were randomly divided into the control group and the model group ,10 cases in each group The gel state Aβ(10μg) was injected into the rat′s bilateral hippocampus in the model group ,while the control group was injected with normal saline ;the Morris water maze test was performed to assess the rat′s learning and memory ability ;the thio‐nine stain was used for observing the morphology and quantity of cerebral cortex neurons ;the enzyme linked immunosorbent assay (ELISA) was adopted to detect the serum tumor necrosis factor alpha (TNF‐α) and interleukin 1 beta(IL‐1β) levels ;the immuno‐histochemical was used to detect the expression of glial cell and glial fibrillary acidic protein (GFAP) .Results The various indexes in the model group were significantly lower than those in the control group(P<0 .05);the quantity of cerebral cortex neurons in the model group was significantly decreased compared with that in the control group (P<0 .01);the ELISA results showed that the lev‐els of TNF‐αand IL‐1βin the model group were significantly higher than those in the control group(P<0 .05);the immunohisto‐chemistry showed that the number of GFAP positive cells in the hippocampus in the model group was significantly more than that in the control group(P<0 .05) .Conclusion Aβmight activate the glial cells and promote the release of inflammatory cytokines ,which causes the damage of rat cerebral neurons and leads to decrease the rat′s learning and memory ability .

Polyethylene glycol (PEG) is extensively used to increasing the in vivo and in vitro stability of liposomes. However, PEGylated liposomes also produce some negative effects with further research, such as low cellular uptake, poor "endosomal escape" of pH sensitive liposome (PSL) and accelerated blood clearance (ABC) phenomenon, and this situation is referred as the "PEG dilemma". "PEG dilemma" posed severe challenges for the targeted delivery of PEGylated liposomes-loaded anticancer drugs, effective intracellular release of PEGylated PSL-encapsulated gene and protein drugs, and repeated administration of PEGylated liposomes. Therefore, it is urgent to solve the "PEG dilemma". This review focused on the definition, classification of "PEG dilemma", and discussed several possible approaches to overcome "PEG dilemma".
Antineoplastic Agents ; chemistry ; Drug Carriers ; chemistry ; Liposomes ; chemistry ; Polyethylene Glycols ; chemistry

Objective To explore the histocompatibility and plasticitity of hydroxyapatite/high-density polyethylene composite (HMPE). Design Experimental study. Participants Forty-five rabbits. Methods Rabbits were randomly devided into HA/PE, HA and PE surgical groups, every group included 15 rabbits. Osteoeetomy of orbital rim and implantation (HA/PE, HA and PE materials were used) were performed on 3 groups of the rabbits. At 1st week, 4th week, 8th week, 12th week and 24th week of postoperation, we took out the samples from the rabbits separately in each group and did histopathologic and electron microscope examinations. Main Outcome Measures Physical signs of the implant combination with the bone. The bone cell appeared in the implants. Results After 12 weeks, the calcium salts deposit like bone plate with some osteoblast inside the HA/PE implants was observed. After 8 weeks, the calci- um salts deposit inside the HA plants was observed. In the PE implants, nothing could be seen but some fibrous connective tissue. Conclusion HMPE is an ideal repair material especially for orbital bone.

Objective:To investigate the efficacy of sequential therapy of CD19 and CD22 chimeric antigen receptor T cells (CAR-T) for relapsed/refractory mediastinal B lymphoblastic lymphoma (B-LBL).Methods:One patient with relapsed/refractory mediastinal B-LBL treated with sequential therapy of CD19 and CD22 CAR-T who was admitted to Tongji Hospital of Huazhong University of Science and Technology Tongji Medical College in March 2017 was reported. At 1, 3, 6, 12 and 18 months after CAR-T therapy, the indicators of primary disease remission were monitored and the relevant literature was reviewed.Results:The patient relapsed and progressed after third-line chemotherapy, and then received sequential therapy of CD19 and CD22 CAR-T. In the course of cellular immunotherapy, the patient presented grade 1 cytokine release syndrome. After active treatment, the patient got stable condition and was discharged. The patient came to the hospital for regular review, and the mediastinal mass of the patient was dynamically followed up. After CAR-T therapy, the mediastinal mass of the patient was significantly reduced, and the patient was in continuous remission for 18 months.Conclusions:Sequential therapy of CD19 and CD22 CAR-T provides a new therapeutic approach for relapsed/refractory B-LBL. For patients with poor curative effect of conventional chemotherapy, CAR-T therapy should be actively performed as soon as possible to improve the remission rate and the long-term prognosis of patients.