Objective:To clone human CGI-100 gene and reconstruct its eukaryotic expressive vector for further investigation.Methods:The full coding domain sequence of human CGI-100 gene was cloned from human leukemic K562 cells with RT-PCR and was sub-cloned into pMD18-T Simple vector.After confirmed by DNA sequencing,the targeted DNA fragment,digested with BamHⅠand PstⅠ,was directionally cloned into eukaryotic expressive plasmid pIRES2-EGFP,then,the reconstructed plasmid was identified with enzyme digestion,PCR and sequencing.Results:It was demonstrated that the full coding domain sequence of human CGI-100 gene was accurately cloned into digestion sites between BamHⅠand PstⅠin pIRES2-EGFP without mutation and transposition.Conclusion:The reconstruction and verifying of eukaryotic expressive plasmid containing CGI-100 gene are successful,which establishes the foundation of further investigation.

Objective To explore the use of Ultrasonic Biomicroscopy(UBM)for peri-operative examination of traumatic cyclodialysis.MethodsUBM was used in 33 eyes of 33 cases who were diagnosed as traumatic cyclodialysis.The morphologic characters were observed peri-operatively.ResultsCiliary detachment of 360° was verified in the 33 eyes with cyclodialysis of more than 2 clock hours.1 month after cyclopexy,the space of ciliary detachment disappeared or diminished and all reattached 3 months after operation.ConclusionUBM is safe and effective for checking traumatic cyclodialysis.

Objective To study the process of brain protein hydrolysate of inactivate and virus removal.Methods The Parvoviridae parvovirus genera of porcine parvovirus (PPV),vesicular stomatitis virus rhabdovirus genera of vesicular stomatitis virus (VSV) were chosen as a model virus,wherein PPV represents no envelope deoxyribonucleic acid(DNA) virus,VSV represents the envelope ribonucleic acid(RNA) virus.Simulation of the production process of virus inactivation steps 100 ℃ × 30 min,ultrafiltration as inactivation/removal condition.The virus respectively according to 1 ∶ 9 into the brain protein hydrolysate,high temperature and ultra filtration virus inactivation/removal.In pig kidney cells (PK-15) in PPV cell culture,Africa green monkey kidney cells(Vero cells) cultured VSV,determination of virus titer.Results PPV and VSV through the sterilization,virus median tissue culture infective dose(TCID50) were 6.15log/0.1mL(logs),5.37 log/0.1mL(logs) ;removal processaverage virus reduction coefficient were 6.15log/0.1mL(logs),5.37 log/0.1mL(logs).Conclusion The high temperature and ultra filtration produces brain protein hydrolysate solution process are effective virus inactivation/removal process.

We use PBL teaching mode in experimental teaching of clinical hematologic laboratory.By this method,the knowledge can be mastered by the students more efficiently and the analyzing ability was also improved.We hope that these experiences can be useful in our country's clinical laboratory teaching.

Objective To establish a new multiplex-PCR assay to improve the detection rate of mutations in the DMD gene in Chinese patients. Methods A retrospective review of DMD deletion spectrum of 355 DMD patients with deletions all over the gene was performed. All deletions were confirmed by " one-step approach" diagnostic procedure and MLPA analysis. The exons with high frequency of mutations were identified to constitute the amplification system and the PCR conditions were optimized. Results Two new multiplex-PCR assays were established. Assay one was used to detect 10 exons including exon 5, 8, 17, 44, 45, 47, 49, 50, 51 and 52 of DMD gene, in two PCR sets. The theoretical detection rate would be 92% (326/355). Assay two was used to detect 5 exons including exon 12, 19, 35, 43 and 54, which could be used to screen additional 5% (17/355) deletion cases. The method was validated in other 22 DMD patients. Multiplex-PCR results were completely identical to the MLPA results in all 22 DMD patients. Conclusions The two multiplex-PCR assays were established based on the analysis of 355 Chinese DMD patients with gene deletions. It is believed that the new approach would be more applicable for deletion detection on the Chinese DMD patients since the DMD cases involved were from the whole country.

BACKGROUND:Previous studies discovered that pRST98, original y isolated from Salmonel a enterica serovar typhimurium (S.typhimurium) could promote bacterial biofilm formation. In addition, bacterial harboring pRST98 can promote the secretion and expression of interleukin-10 after infection in cells and animals.
OBJECTIVE:In vitro studies have discovered the effects of interleukin-10 at varying concentrations and conjugative plasmid pRST98 on the biofilm formation of S.typhimurium.
METHODS:S.typhimurium wild-type strainχ3306, virulence plasmid-deletion S.typhimurium strainχ3337 and pRST98-transconjugant S.typhimuriumχ3337/pRST98 were established in vitro and cultured for biofilm formation. 1, 10, 100 μg/L interleukin-10 were added during the biofilm formation. 0 μg/L interleukin-10 was set as a control. Crystal violet staining method, semi-quantitative method, confocal laser scanning microscopy and scanning electron microscopy were used to determine the effects of interleukin-10 on the biofilm formation and compare the effects of S.typhimurium with or without pRST98.
RESULTS AND CONCLUSION:Intra-group comparison showed that, compared with the control group, S.typhimurium gathered together and formed thicker biofilm in concentration of 1 and 10 μg/L of interleukin-10. The promotion effects of S.typhimurium on biofilm formation were greatly improved in 10 μg/L. Interleukin-10 in 100 μg/L inhibited S.typhimurium biofilm formation. Inter-group comparison showed that, A570 inχ3337/pRST98 was greatly higher than that inχ3306 andχ3337 under the same concentration of interleukin-10. The results indicate that both 1 and 10 μg/L of interleukin-10 promote biofilm formation, especial y bacteria harboring pRST98.

Objective To observe hemodynamic changes in patients with primary hypertension with wave intensity (WI). Methods Carotid arteries of 36 patients with primary hypertension and 30 age-matched normal controls were examined with imaging technique of WI. The following parameters were measured: the first wave peak in early ejection (W1), the second wave peak in late ejection (W2), the negative area during the mid-ejection (NA), the interval between the R wave of the ECG and the first peak of W1 (R-1st), the interval between the first peak and the second peak (1~(st)-2~(nd)), the ratio of R-1st and a cardiac cycle time R-1_(HR)~(st)) and the ratio of 1~(st)-2~(nd) , as well as one cardiac cycle time (1~(st)-2(_(HR)~(nd)). Results ①W1 in primary hypertension group increased compared with those of normal controls (P＜0.01), while no significant difference of W2, NA, R-1st, 1~(st)-2~(nd), R-1_(HR)~(st), 1~(st)-2_(nd)~(HR) was detected (P＞0.05). ②Both W1 and W2 correlated positively with pulse pressure (PP) and systolic blood pressure (SBP) (r=0.66, 0.55, P＜0.01;r=0.62, 0.44, P＜0.01). W1, W2 and age, DBP were not related significantly (P＞0.05). Conclusion The hemodynamic parameters of WI technology provide a new way to evaluate the dynamics of the heart and vascular system and their interaction.

Objective To develop a rapid,reliable and convenient approach for diagnosing the homozygous deletion of SMN1 gene.Methods SMN1 gene was amplified specifically with double allele-specific PCR(AS-PCR).Meanwhile,one inrelevant gene was amplified as internal control by PAGE and agarose gel electrophoresis analysis to determine whether the sick children were with homozygous deletion of SMN1 genes.Results The homozygous deletion of exon7 in SMN1 gene was identified by agarose gel electrophoresis or PAGE accurately.Conclusion Compared to PCR-RFLP and DHPLC used in the past,this approach can diagnose homozygous deletion of SMA much more accurate,easier and more convenient without completed following analyses.

Objective:To examine the validity and reliability of the Chinese version of the Body Acceptance by Others Scale-2(BAOS-2)in adults.Methods:Totally 616 adults(aged 18-56 years)were selected to test the structural validity,and internal consistency reliability of the scale.The Body Appreciation Scale-2(BAS-2),Func-tionality Appreciation Scale(FAS),Self-Compassion Scale(SCS),Self-Esteem Scale(SES)and Satisfaction with Life Scale(SWLS)were used as criteria to test criterion validity.A sample of 55 adults was retested 2 weeks later for the test-retest reliability.Results:The exploratory factor analysis extracted one factor,and the factor loading range of each item was 0.59-0.76.The confirmatory factor analysis showed that the factor model fit indices were acceptable(x2/df=2.99,CFI=0.91,GFI=0.90,TLI=0.89,RMR=0.04,RMSEA=0.08).The scores of the Chinese version of BAOS-2 were positively correlated with the scores of BAS-2,FAS,SCS,SES and SWLS(r=0.43-0.66,Ps＜0.001).The Cronbach a of the Chinese version of BAOS-2 was 0.90,and the test-retest reliabili-ty(ICC)was 0.65.Conclusion:The Chinese version of the Body Acceptance by Others Scale-2(BAOS-2)has i-deal validity and reliability.

Recently, some global cases of 2019 novel coronavirus (COVID-19) pneumonia have been caused by second- or third-generationtransmission of the viral infection, resulting in no traceable epidemiological history. Owing to the complications of COVID-19pneumonia, the first symptom and imaging features of patients can be very atypical and early diagnosis of COVID-19 infectionsremains a challenge. It would aid radiologists and clinicians to be aware of the early atypical symptom and imaging featuresof the disease and contribute to the prevention of infected patients being missed.