Objective To compare the purification process of two types of ceramic hydroxyapatite(CHT I and CHT II)to remove the aggregates from two monoclonal antibodies(mAb 1 and mAb 2).Methods All the chromatography runs were performed on AKTA AVANT 150 with Tricon 10/50 column.The dynamic binding capacity( DBC) of two types of CHT was studied firstly, and then purification research was carried out selecting the suitable DBC.The column was equilibrated with 5 mmol/L sodium dihydrogen phosphate pH 6.5, and then was eluted with gradient buffers which were 10 mmol/L sodium dihydrogen phosphate pH 6.5 and 2 mol/L sodium chloride pH 6.5.Aggregate content in loading and elution pool was evaluated by size exclusion chromatography.Scale-up process was carried on 20 cm height chromatography column XK16/40.Results DBC of CHT I for mAb 1 was 40 mg/mL and mAb 2 was 45 mg/mL.After purity, monomer content of mAb 1 reached 98.6% and yield was 92.5% and monomer content of mAb 2 reached 98.8%and yield was 91.5%.DBC of CHT II for mAb1 was 16 mg/mL and mAb 2 was 20 mg/mL.After purity, monomer content of mAb 1 reached 99.8% and yield was 91.8% and monomer content of mAb 2 reached 99.9% and yield was 92.2%.Conclusion Two types of CHT both can remove aggregates effectively from monoclonal antibodies when aggregate content reaches more than 10%, and results conform to the regulations.CHT I has higher dynamic binding capacity than CHT II, and CHT II is superior to CHT I in removing aggregate efficiency.The purification process is simple and can be easily scaled up in pilot and manufacture.Therefore, it meets the requirement pilot and scale production.

Academic Inspection is not only an important part of clinical teaching,but also a vital tache of improvement for medical treatment quality.Unlike internal medicine and surgery,acdemic inspection of ICU has its own characteristics.PBL teaching method on acdemic inspection of attending doctor in ICU is explored in this article.

Background Experimental autoimmune uveitis (EAU) is a common animal model of uveitis.Natural killer (NK) cells have been confirmed to be a type of strong inflammation-causing cells,but its role in EAU is still studing.Objective This study was designed to explore the role and mechanism of NK cells in the pathogenesis of EAU.Methods Thirty-six SPF Lewis rats were randomly divided into expeimental control group and EAU 6-,9-,12-,16-,and 21-day groups (6 rats for each group).Rats in EAU group received subcutaneous injection interphotoreceptor retinoid binding protein (IRBP) combining 5 mg/ml tubercle bacillus with complete Freund's adjuvant (CFA) emulsion in foot pads,and then 400 ng pertussis toxin was intraperitoneally injected to extablish EAU models in the EAU 6-,9-,12-,16-,and 21-day group,and normal saline solution combined with CFA and 400 ng pertussis toxin was used in the same way in the experimental control group.The inflammatory response was observed by slit lamp daily after modeling and scored based on Caspi criteria.The eyeballs were extracted in 6,9,12,16 and 21 days after modeling for retinal histopathological examination,Immunofluorescent double-staining was employed to detect and locate the expression of NK cells in the retina.In addition,25 model rats were divided into EAU 0-,3-,6-,9-and 12-day groups,with 5 rats for each group,and eyeballs were extracted to prepare tissue homogenate.The expression of CXCL10 mRNA,and CXCL12 mRNA NK cell chemokines,in the tissue homogenate was assayed by real-time quantitative PCR.The use and care of the rats followed Regulations for the Administration of Affair Concerning Experimental Animal by State Science and Technology Commission.Results No inflammatory sign in ocular anterior segment of the rats was seen in the experimental control group.The expansion of rat iris vessels was found in the EAU 6-day group,and exudes and hypopyon of the anterior chamber occurred in the EAU 9-day group and the inflammation peaked in the EAU 12-day gorup.The rat retinal structure was normal in the experimental control group,and the arrangement disorder of retinal structure,the cell separation in outer nuclear layer and damage of photoreceptors were found under the optical microscope in different degree in various EAU groups,with the most serious change in the EAU 12-day group.Immunofluorescent double staining showed normally arranged nucleus in the experimental control group,and a lot of NK infiltration was seen in the EAU 6-day group and peaked in the EAU 9-day group.The expression level of CXCL10 mRNA in the EAU 9-day group was 34.298 ± 16.689,which was significantly higher than that in the EAU 3-,6-and 12-day group,respectively (1.390 ± 0.660,3.359 ± 2.581,4.711 ±1.387) (all at P＜0.01).No significant differences were found in the relative expression of CXCL12 mRNA among different EAU groups (F=2.851,P＞0.05).Conclusions Retinal NK cell infiltration occurs in the early stage of EAU,and the severity of NK cell infiltration is consistent with the inflammatory process and CXCL10 expression,suggesting NK cells play an important role in the early stage of EAU,and CXCL10 is an important chemokine of NK cells in EAU rats.

OBJECTIVE: To analyze the relationship of the height difference and bone density (BD) of premenopausal and postmenopausal women. METHODS: The height values of 191 premenopausal and postmenopausal women were recorded, and the BD values of lumbar vertebrae and hip were detected by double energy X-ray BD detector. RESULTS: The lower the height of the postmenopausal women, the less the BD value. The BD of lumbar vertebrae dropped 0.025 5 g/cm(2) with each 2 cm of the shortened height, and the BD of hip joint dropped 0.029 2 g/cm(2). The shortened value in postmenopausal women with osteoporosis was statistically greater than that in postmenopausal women without osteoporosis. CONCLUSION: The BD of the postmenopausal women can be estimated by the calculation of their shortened height value.

Objective To screen out some Chinese medicines with obvious inhibitory actions on triple-negative breast cancer based on data mining. Methods Literatures of breast cancer effectively treated by Chinese medicine issued in recent 15 years were collected, and then the frequency analysis and cluster analysis were used to screen the frequently-used herbs. According to the above results, the serum containing the obtained drugs were prepared respectively. And then the proliferation, migration and invasion abilities of breast cancer cells MDB-MB-231 treated with the serum containing various Chinese medicines for 12, 24 h were observed by methyl thiazolyl tetrazolium(MTT) assay and transwell cell invasion and migration experiments, respectively. Results The frequently-used herbs were screened out as Rhizoma Chuanxiong, Herba Taraxaci, Herba Hedyotis Diffusae, Radix Trichosanthis, Radix Achyranthis Bidentatae. The proliferation of MDB-MB-231 cells was significantly inhibited by serum seperately containing Rhizoma Chuanxiong, Herba Taraxaci, Herba Hedyotis Diffusae, Radix Trichosanthis, and Radix Achyranthis Bidentatae to different degrees, and the migration abilities of MDB-MB-231 cells were obviously inhibited by serum containing Herba Taraxaci, Herba Hedyotis Diffusae, Radix Achyranthis Bidentatae, respectively. And the invasion abilities of MDB-MB-231 cells were obviously inhibited by serum containing Herba Taraxaci, Rhizoma Chuanxiong, Herba Hedyotis Diffusae, respectively. Conclusion Chinese medicines such as Rhizoma Chuanxiong, Herba Taraxaci, Herba Hedyotis Diffusae, Radix richosanthis, Radix Achyranthis Bidentatae obtained by data mining technology have obviously inhibitory effects on triple-negative breast cancer.

AIM:To investigate the mechanism of apoptosis during the process of adult rat mesenchymal stem cells(MSCs)differentiating into neuron-like cells in vitro.METHODS:The MSCs were isolated primarily from adult rats bone marrow by density gradient and then expanded in medium as undifferentiated cells for 3-5 generations.The MSCs were divided into three groups at random.The control group was induced with ?-mercaptoethanol(?-ME).Meanwhile,the U0126 group was given ?-ME and U0126(10 ?mol/L)added at the beginning of induction.The PMA groups were treated with ?-ME and different concentrations of PMA since pre-induction.The effects of U0126 and PMA on the activity of neuron-like cells were observed by MTT.The effects of U0126 and PMA on the expression of neuron specific marker nestin and expression of apoptosis-related protein caspase,Bcl-2,Bax in neuron-like cells were detected by using immunocytochemistry method.TUNEL technique was used to detect apoptosis index.RESULTS:Compared to control group,neuron viability,nestin and Bcl-2 increased and neuron apoptosis decreased in U0126 group(P

Objective To explore the relationship of personality and coping style between parents and male inpatients with internet addiction disorder. Methods 51 male inpatients with internet addiction disorder and their parents were investigated with Cattell 16PF and Coping Style Questionnaire to evaluate their personality and coping style. All data were analyzed by partial correlate analysis. Results Multi-factors of 16PF( warmth, reasoning, emotional stability, dominance, vigilance, sensitivity, abstractedness, privateness, openness to change, perfectionism, apprehension, tension ) of male inpatients were significantly correlated with father' s ( r = 0.72 ～ 0. 85 or - 0. 71 ～ - 0.93 ) or mother' s 16PF( r = 0.73 ～ 0.92 or - 0. 72 ～ - 0.81 ), bu+ non of those factors corrected with the same factor of parental. Male inpatients' coping style did not corrected with father' s, but their problem sloving, rationalization and self-accusation corrected with mother's seeking support, rationalization and avoidance respectively. Conclusion The personality of male inpatients with internet addiction disorder is affected by parental personality, but under the impact of internet addiction disorder, this kind of influence is weakened. In coping style, the father' s influence is far weaker than the mother' s, suggesting that there is father function flaw during the process of those inpatients growing up.

Objective To investigate the relationship between human papilloma virus(HPV)infection and the development of laryngeal squamous cell carcinoma(LSCC).Methods To elucidate the role of HPV in the development of LSCC,we employed polymerase chain reaction(PCR)based on four pairs of primers an4 in situ hybridization(ISH)to screen the HPV infection in 84 ISCC tissues.Results Using HPV L1 general primer amplification,HPV DNA was detected in 23(27.4%)of the 84 LSCC samples.However,when specific primers for HPV-16 or-18 were used to amplify E6 and E7 in all samples,29 cases(34.5%)were positive for HPV-16,while 6 cases(7.1%)were positive for HPV 18.Coinfeetion of HPV-16 and-18 were found in 4cases (4.8%).Overall,HPV type 16 and 18 infections were present in 36.9% of the LSCC samples.In addition,the positive rate of HPV 16 E6 mRNA was 30.9%(26/84)in LSCC by ISH with digoxin-labeled sense probes of HPV 16 E6.Conclusion High-risk HPV-16may be an etiologic factor in the development of laryngeal squamous cell carcinoma, while the complicated molecular mechanism of HPV16 inducedtumorgenesis needs a further study.

Background Dacryocystitis is one of the most common infectious eye diseases.The gold standard for the identification of bacteria causing dacryocystitis is bacterial culture.The combination of regular culture method with molecular biology techeniques will generate more reliable results.However,very few research data are available in ophthalmological studies in this area.Objective This study was to identify the genera and species of the dacryocystitis-causing bacteria by PCR amplification of the 16S rRNA sequences.Methods Ten cases of qualified standardized bacteria samples were taken,and the nucleic acids were released in the heating process of the PCR procedure.The 16S rRNA genes were amplified and sequenced,and the genera and species were identified using BLAST from GenBank,and the results were used to compare with the results from biochemical identification to test the reliability of this method.The cultured bacterial species from the lacrimal sac secretions from 30 cases of dacryocystitis patients were identified with the above method.Results The outcome of the PCR identification for the 10 cases of quality control standard bacterial specimens was consistent with the results from the biochemical identification.The identification of the 30 cases of dacryocystitis through sequencing the 16S rRNA revealed there were 13 cases of Staphylococcus epidermidis infection,2 cases of Staphylococcus warneri infection,1 case of Staphylococcus hominis infection,5 cases of Corynebacterium macginleyi infection,3 cases of Streptococcus pneumonia infection,2 cases of Bacillus cereus infection,1 case of Micrococcus luteus infection,1 case of Moraxella catarrhalis infection,1 case of Moraxella osloensis infection and 1 case of Pseudomonas aeruginosa infection.Conclusions Sequencing the 16S rRNA is an accurate and specific way for the identification of the genera and species of bacteria that cause dacryocystitis in patients.This sequencing method is feasible in monitoring a variety of dacryocystitis-causing pathogens.More information and epidemiological statistics about dacryocystitis can be obtained from 16S rRNA sequencing.

Objective To investigate the effect of anterior percutaneous endoscopic discectomy (APECD) and open-door laminoplasty (ODLP) through hybrid surgery in the treatment of multisegmental cervical stenosis and giant disc herniation. Methods This study involved 3 patients with multisegmental cervical stenosis and giant disc herniation confirmed by MRI. Among them, there were 2 males and 1 female, with ages from 56-61. All patients showed significant paresthesia or weakness, and were treated between September and November 2016. The surgery was performed by first the ODLP that made spinal cord back shift, and then APECD for the second step. The visual analog scale (VAS) and neck disability index (NDI) were assessed before and after operation. Results The VAS and NDI scores were improved two weeks after operation. No adverse events like spinal cord injury and vascular injury were found during the operation. After operation, no patients were found incision infection, hematoma formation, cerebrospinal fluid leakage, dysphagia, trachyphonia and so on. Conclusion The hybrid surgery of APECD and ODLP for the treatment of the multisegmental cervical stenosis and giant disc herniation can not only decompress the nerve safely and improve the function, but also preserve cervical intervertebral disc and motion segments, therefore delaying the degeneration of adjacent segments with clinical significance.