Objective To discuss the role of DNA-dependent protein kinase catalytic subunit (DNA-DPKCS) in human lung adenocarcinoma cell line (A549) by using small interfering RNA (siRNA) to specifically knockdown DNA-DPKCS expression and its effects on cell proliferation, cell cycle and radio-sensitivity. Methods The DNA-DPKCS-siRNA expression vector was constructed and transfected into A549 cell line. The transformed clones were randomly selected and isolated. The cell cycle distribution and apop-tesis were analyzed by flowcytometry analysis. Cell survival was detected by using clonogenic formation as-say. Results With specific inhibition of DNA-DPKCS expression, stable transfected cell line 549pRNA-DNA-DPKCS was constructed by RNA interference technique. The 549pRNA-C and 549pSUPER cell lines were the control cell lines tansfected with control and blank plasmids, respectively. Compared with A549 cells, the expression levels of DNA-DPKCS mRNA (0.110: 1. 000), protein (0. 870: 2.967) and activity of DNA-DPKCS (0.004: 0.266) in 549pRNA-DNA-DPKCS cells were significantly lower (F = 80.55 ,P < 0.01;F=63.96, P<0.01;F=51.62,P<0.01, respectively). The analysis of SF_2(0.25:0.76), D_0 (1.42:1.62) and D_q (0.06: 1. 00) showed significant difference between 549pRNA-DNA-DPKCS and A549 cells (F = 996.86, P < 0.01 ; F = 17.41, P < 0.05 ; F = 68.92, P < 0.01). The number of 549pRNA-DNA-DPKCS cells in S (24.5%: 35.5%) and G_2 (10.7%: 11.0%) phases was significantly decreased (F = 4.83, P<0.05 and F=32.04, P <0.01, respectively). Conclusions In A549 cells, inhibit of DNA-DPKCS gene expression can enhance the radiosensitivity and affect cell cycle distribution.

AIM:To construct recombinant adenovirus vector carrying human miR-133a and study its expression in human mesenchymal stem cells(hMSCs).METHODS:The PCR product containing miR-133a was amplified from human genomic DNA and inserted into the adenoviral shuttle vector pAdTrack-CMV.Then the recombinant shuttle plasmid linearized by pmeⅠwas cotransformed into competent E.coli.BJ5183 with the adenoviral backbone plasmid pAdEasy-1 to generate the recombinant adenovirus vector rAd-mir-133a.rAd-mir-133a was then packaged and amplified in human embryonic kidney 293(HEK293) cells.The purified rAd-miR-133a was used to infect the hMSCs and the expression of miR-133a was detected by non-quantitative RT-PCR and real-time PCR.RESULTS:The recombinant adenovirus shuttle vector pAdTrack-CMV-miR-133a was constructed and verified by restriction endonuclease analysis and DNA sequence analysis.rAd-miR-133a was successfully packaged and amplified in HEK293 cells.The transcriptions of primary miR-133a and mature miR-133a were over-expressed in the hMSCs infected with rAd-miR-133a.CONCLUSION:The recombinant adenovirus vector carrying human miR-133a is successfully constructed,which lay a foundation for miR-133a function study.

Background and objective All the advanced NSCLC patients that received EGFR-TKI therapy will eventually relapse after a period of efficacy. The aim of this study is to investigate the serum biomarkers as potential predictive factors for the efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) targeted therapy in advanced non-small cell lung cancer. Methods Twenty serf-paired serum samples were collected from 9 advanced NSCLC patients that evaluated as disease control (SD or PR) after gefinitib therapy, at the time points of before and after gefinitib treatment but 2 weeks before being evaluated as disease progress. AII samples were pre-separated by WCX microbeads, and then detected on the MALDI-TOF-MS platform of Bruker AutoflexTM. ClinProTools (Version: 2.1) was used to analyze the differentially expressed proteins. Results There were 7 protein peaks (m/z), 3 242.09, 8 690.36, 2 952.64, 3 224.04, 1 450.51, 1 887.8 and 3 935.73 found statistically differentially expressed between the self-paired samples. Three proteins (3 242.09, 2 952.64 and 3 224.04) were down-regulated and four proteins (8 690.36, 1 450.51, 1 887.8 and 3 935.73) up-regulated in gefinitib treated sera. Conclusion The data here suggest that several specific protein peaks might indicate gefinitib resistance, yet the identities of these proteins and the mechanisms underlying the responsiveness to gefinitib treatment need further investigation.

Objective To assess the efficacy and safety of nebulized hypertonic saline solution in infant with bronchiolitis.Methods From January 2014 to January 2016,95 patients at 3-13 months old in our hospital who diagnosed as bronchiolitis were randomly divided into three groups.On the basis of conventional suit support treatment,the observation group A was given nebulized 3％ hypertonic saline (n =32),the observation group B was given nebulized 3％ hypertonic saline and salbutamol (n =32),and the control group C was given nebulized normal saline (0.9％) and salbutamol (n =31),this therapy was repeated every 6 hours until discharge.The Lowell score,clinical symptoms remission time,days of hospitalization and incidence rate of adverse reaction of the three groups were compared.Results After treatment,the symptoms and signs of the three groups were all improved.The Lowell scores were lower in the two observation groups compared to the control group C in 24,48,72 hours after treatment[(5.81 ±1.53) points and (5.85 ± 1.37) points vs.(6.61 ± 1.54) points,(4.75 ± 1.34) points and (4.72 ± 1.30) points vs.(5.52 ± 1.29) points,(3.19 ± 1.15) points and (3.22 ± 1.16) points vs.(3.90 ± 1.01) points,Z =-1.999,-2.241,-2.518 and-2.002,-2.335,-2.316,all P ＜ 0.05).And the cough,wheezing remission time and pulmonary rales disappearance time,days of hospitalization in the two observation groups were also shorter,there were statistically significant differences [(6.63 ± 1.41) d and (6.56 ± 1.37) d vs.(7.35 ± 1.25) d,(5.19 ± 1.03) d and (5.25 ± 1.05)d vs.(5.87 ± 1.09)d,(5.75 ±1.34)d and (5.72 ± 1.51)d vs.(6.68 ± 1.60)d,(7.25 ± 1.37)d and (7.16±1.48)d vs.(8.10±l.47)d,Z=-2.498,-2.469,-2.359,-2.213 and-2.982,-2.405,-2.373,-2.222,P ＜0.05,or P ＜0.01)].There were no significant differences in the Lowell score,the length of time of cough,wheeze,lung rales disappears and the length of hospital stay between the observation group A and observation groupB[(5.81 ± 1.53) points vs.(5.85 ± 1.37) points,(4.75 ± 1.34) points vs.(4.72 ±1.30) points,(3.19± 1.15) points vs.(3.22 ± 1.16) points,(6.63 ± 1.41) d vs.(6.56 ± 1.37) d,(5.19 ±1.03)d vs.(5.25 ± 1.05)d,(5.75 ± 1.34) d vs.(5.72 ± 1.51) d,(7.25 ± 1.37) d vs.(7.16 ± 1.48) d,Z =-0.164,-0.021,-0.140,-0.295,-0.167,-0.374,-0.233,all P ＞ 0.05].Children in three groups had no serious adverse events (all P ＞ 0.05).Conclusion Nebulized hypertonic saline in the treatment of bronchiolitis can relieve symptoms and signs,shorten the hospitalization time,and has less adverse reaction,it is worthy of clinical use.

Objective:To evaluate the diagnostic accuracy and application value of high-frequency ultrasonography in proximal radial nerve injuries associated with humeral shaft fractures.Methods:A retrospective case series study was performed for 19 patients with proximal radial nerve injuries associated with humeral shaft fractures treated in Huashan Hospital Affiliated to Fudan University from August 2014 to September 2020. The were 17 males and 2 were females，with the age range of 16-55 years ［（38.1±12.7）years］. Of all，11 patients were injured at the left side and 8 at the right side. All the patients had histories of traumatic humeral shaft fracture and were treated in other hospitals，including internal fixation in 17 patients，external fixation in 1 and internal fixation combined with external fixation in 1. All patients underwent radial nerve exploration surgeries，among which 8 had high-level radial nerve release and 11 had high-level radial nerve suture or graft transplantation. All patients underwent high-frequency ultrasound examination before surgery. The ultrasonographic characteristics of the radial nerve were recorded，including the continuity of the epineurium，honey-comb structure on the transversal section，neuromas on the longitudinal section and external scar or callus or metal fixation compressing the nerve. The injury type and neural continuity of each radial nerve were evaluated. The radial nerves of type Ⅰ，type Ⅱ，type Ⅲa ，type Ⅲa ，type Ⅲb and type Ⅳ appeared as normal，swelling，short-segment compressed，neuroma-like and ruptured，respectively. Taking the intraoperative findings as the gold standard，the diagnostic coincidence rate，sensitivity，specificity，positive predictive value and negative predictive of high-frequency ultrasound were analyzed in diagnosis of proximal radial nerve injuries associated with humeral shaft fractures.Results:According to ultrasonographic characteristics，proximal radial nerve injuries were classified into 4 types in 5 subtypes. It was confirmed by surgeries that there were 1 patient with type Ⅱ，4 with type Ⅲa，1 with type Ⅲb and 13 with type Ⅳ，with no type Ⅰ. The diagnostic coincidence rate of high-frequency ultrasound was 89%. The diagnostic coincidence rate of type Ⅱ，type Ⅲa，type Ⅲa，type Ⅲb and type Ⅳ was 100%，100%，100% and 85%，respectively. The sensitivity of high-frequency ultrasound for evaluating the neural continuity was 75%，the specificity was 100%，the positive predictive value was 100%，the negative predictive value was 85%.Conclusions:The high-frequency ultrasound has a relatively high sensitivity and specificity in diagnosis of proximal radial nerve injuries associated with humeral shaft fractures，which can provide relatively accurate morphological information for clinical diagnosis and treatment.

Objective To investigate the differences in the expression of PHF20 and Bax and their correlations in non-small cell lung cancer before chemotherapy.Methods An immunohistochemical method was used to detect the expression of PHF20 and Bax in non-small cell lung cancer,and to analyze the clinical significance of PHF20 and the possible correlation between PHF20 and Bax proteins.Results PHF20 protein is expressed in the cytoplasm of non-small cell lung cancer cells.Moreover,it is highly expressed in squamous cell carcinoma,less expressed in adenocarcinoma,and closely related with cell differentiation,TNM staging,and lymph node metastasis.The expression of PHF20 and Bax was positively correlated with squamous cell lung carcinoma.Conclusion The expression of PHF20 in non-small cell lung cancer is closely associated with tumor progression and the expression of Bax.PHF20 may be a new target for the treatment of non-small cell lung cancer.