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 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 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.

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 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 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 investigate the effects of different lighting(sulfur lamp, heat radiation lamp, fluorescent lamp and LED lamp) on reproductive system in depressive male rats.Methods CUMS depressive model of rats was established through the stimulations of chronic unpredictable mild stress(CUMS), then they were randomly divided into 5 groups:depressive model group, sulfur lamp group, heat radiation lamp group, fluorescent lamp group and LED lamp group.After 45 d of continuous illumination, serum was collected and reproductive organs were removed from rats, then the organ index of testicle,epididymis,seminal vesicle and kidney was calculated, the concentrations of testosterone (T), estradiol (E2), prolactin (PRL) and progesterone (PROG)in the serum were detected by ELISA, the histomorphological lesion of testicle was observed under microscope with hematoxylin-eosin (HE) staining, the expression of melatonin receptors (Mel 1a), 3β-HSD and P450scc of testicle were detected by Western blot.Results The organ index of testicle, epididymis and seminal vesicle, the concentration of T, E2,PRLand PROG, and the expression of Mel 1a, 3β-HSD and P450scc in the depressive rats were significantly lower than that of the controls(P<0.05), and the seminiferous tubules of the testis of depressive rats were atrophied.However, the level of T, E2, PRL and PROG were increased,the cell morphology of the testicle was improved, and the expression of Mel 1a, 3β-HSD and and P450scc were upregulated after the sulfur lamp lighting in the depressive male rats compared with depressive model group (P<0.05).Conclusions The testis structure and functions of depressive male rats are improved by sulfur lamp lighting.

Objective To evaluate the associations of human leukocyte antigen(HLA)-DQ gene with autoimmune polyglandular syndrome(APS),type 1 diabetic mellitus(T1DM) and autoimmune thyroid disease(AITD).Methods Fifteen cases of APS,29 cases of T1DM and 40 cases of AITD were selected as research subjects,while 27 healthy children were selected as controls.The DQA1 and DQB1 alleles were determined by polymerase chain reaction(PCR) and sequence-based typing method.The difference of their frequency in children and adolescents was analyzed.Results Compared with controls,APS and T1DM patients had increased frequency of subjects with DQA1*0301,0501(all P

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.