Construction of a stable TrxR1 knockout HCT-116 cell line using CRISPR/Cas9 gene editing system / 生物工程学报

To investigate the cellular target selectivity of small molecules targeting thioredoxin reductase 1, we reported the construction and functional research of a stable TrxR1 gene (encode thioredoxin reductase 1) knockout HCT-116 cell line. We designed and selected TrxR1 knockout sites according to the TrxR1 gene sequence and CRISPR/Cas9 target designing principles. SgRNA oligos based on the selected TrxR1 knockout sites were obtained. Next, we constructed knockout plasmid by cloning the sgRNA into the pCasCMV-Puro-U6 vector. After transfection of the plasmid into HCT-116 cells, TrxR1 knockout HCT-116 cells were selected using puromycin resistance. The TrxR1 knockout efficiency was identified and verified by DNA sequencing, immunoblotting, TRFS-green fluorescent probe, and cellular TrxR1 enzyme activity detection. Finally, the correlation between TrxR1 expression and cellular effects of drugs specifically targeting TrxR1 was investigated by CCK-8 assay. The results demonstrated that the knockout plasmid expressing the sgRNA effectively knocked-out TrxR1 gene within HCT-116 cells, and no expression of TrxR1 protein could be observed in stable TrxR1 knockout HCT-116 (HCT116-TrxR1-KO) cells. The TrxR1-targeting inhibitor auranofin did not show any inhibitory activity against either cellular TrxR1 enzyme activity or cell proliferation. Based on these results, we conclude that a stable TrxR1 gene knockout HCT-116 cell line was obtained through CRISPR/Cas9 techniques, which may facilitate investigating the role of TrxR1 in various diseases.

Surgical experience and risk factors of 133 cases of carotid body tumor / 中华普通外科杂志

Objective:To summarize the surgical experience of carotid body tumor (CBT)and analyze the risk factors.Methods:The clinical and follow-up data of 133 patients with carotid body tumor undergoing surgery at the First Medical Center of PLA General Hospital from Nov 2005 to Apr 2019 were retrospectively analyzed.Results:All of 142 tumors were successfully resected. No patients died perioperatively. Thirty-three (23.2%) cases underwent simple tumor resection, 82 (57.8%) cases underuent tumor resection combined with external carotid artery ligation, 13 (9.2%) cases did internal carotid artery reconstruction, 10 (7.0%) cases had total or external carotid artery repairment, and 4 (2.8%) cases underwent total or internal carotid artery ligation. There were 53 complications, including 43 cases of cranial nerve injury, with an average operating time of 161min (60-500 min), an average blood loss of 308 ml (20-3 000 ml). The follow-up time ranged from 1 to 162 months. No death occurred during the follow-up period.Conclusions:Tumor size and Shamblin typing are surgical risk factors. Most of ShamblinⅠtype tumor can do simple resection, but type Ⅱ and Ⅲ often need to ligate the external carotid artery. Use of great saphenous vein has a favorable long-term patency rate.

Optimizations of an ELISA-like high-throughput screening assay for the discovery of β-catenin/TCF4 interaction antagonists / 生物工程学报

In canonical Wnt/β-catenin signaling pathway, β-catenin/TCF4 (T-cell factor 4) interaction plays an important role in the pathogenesis and development of non-small cell lung cancer (NSCLC), and it is tightly associated with the proliferation, chemoresistance, recurrence and metastasis of NSCLC. Therefore, suppressing β-catenin/TCF4 interaction in Wnt/β-catenin signaling pathway would be a new therapeutic avenue against NSCLC metastasis. In this study, considering the principle of enzyme-linked immunosorbent assay (ELISA), an optimized high-throughput screening (HTS) assay was developed for the discovery of β-catenin/TCF4 interaction antagonists. Subsequently, this ELISA-like screening assay was performed using 2 μg/mL GST-TCF4 βBD and 0.5 μg/mL β-catenin, then a high Z' factor of 0.83 was achieved. A pilot screening of a natural product library using this ELISA-like screening assay identified plumbagin as a potential β-catenin/TCF4 interaction antagonist. Plumbagin remarkably inhibited the proliferation of A549, H1299, MCF7 and SW480 cell lines. More importantly, plumbagin significantly suppressed the β-catenin-responsive transcription in TOPFlash assay. In short, this newly developed ELISA-like screening assay will be vital for the rapid screening of novel Wnt inhibitors targeting β-catenin/TCF4 interaction, and this interaction is a potential anticancer target of plumbagin in vitro.

Endovascular treatment for patients with spontaneous isolated superior mesenteric artery dissection: single-center experiences / 中华普通外科杂志

Objective:To summarize the midterm to long-term outcomes and experiences of endovascular treatment (ET) of spontaneous isolated superior mesenteric artery dissection (SISMAD).Methods:The clinical data of 31 SISMAD patients from Jan 2011 to Dec 2019 treated with ET was retrospectively analyzed.Results:Successful ET was achieved in 29 patients with a technical success rate of 93.5%. A total of 36 self-expandable bare stents were planted in 28 patients and plain old balloon angioplastry (POBA) was performed in 1 patient. Abdominal pain disappeared within 24 hours in 89.3% of the patients after stenting. The rate of perioperative complication was 3.2%. There was no SMA dissection rupture bleeding, nor perioperative death occurred. The mean follow-up time was 53.5 (range, 6 to 110) months. There was no dissecting aneurysm formation, no SMA rupture and bleeding, and no stent rupture during the follow-up. The post ET 1-year, 3-year, and 5-year free from reintervention rate were 100%, 100%, and 91.7%, respectively.Conclusions:ET for SISMAD is safe and effective with satisfactory perioperative and midterm to long-term outcomes.

Effect of miR-106a mimics on osteoarthritis in rats / 中华风湿病学杂志

Objective:To investigate the effect of miR-106a on osteoarthritis.Methods:Twenty-four S-D female rats were randomly divided into three groupsaccording to the random number table: the sham operation group, the osteoarthritis (OA) group and the miR-106a mimic group. Eightweeks after operation, all rats were killed and articular cartilage was separated from the medial tibial plateau of each rat. Histopathology was used to observe the morphological changes and denatured quantity of chondrocytes, the level of inflamm-atory cytokines [interleukin (IL)-1, IL-6, tumor necrosis factor (TNF)-β 1], the expression of apoptosis receptor p65, DR6 protein and the expression of mir-106a RNA. The mean between groups was compared by one-way analysis of variance (ANOVA), and the least significant difference (LSD)- t test was used for the comparison between the two groups. Results:Histopathological staining results showed that the articular surface and synovium of rats in the sham operation group were intact, the chondrocytes were arranged horizontally, and the articular cartilage edge was smooth.In the OA group, the articular cartilage edge was seriously damaged and the arrangement of chondrocytes was disordered.In the miR-106a simulation group, the cartilage structure tended to be normal, occasionally uneven, and the articular cartilage surface was not smooth. Compared with sham operation group, the expression of miR-106a in OA group was significantly decreased ( F=918.02, P<0.01); the expression of inflammatory mediators (IL-1, IL-6, TNF-β) in OA group was also signifi-cantly increased ( F=41 914.86, P<0.01; F=64 85.16, P<0.01; F=8 873.31, P<0.01). The expression levels of DR6 and p65 in OA group were higher ( F=2 319.338, P<0.01; F=1 253.882, P<0.01). Compared with OA group, the levels of inflammatory mediators (IL-1, IL-6, TNF-β) in miR-106a mimetic group were significantly decreased (1.270±0.020, 6.040±0.170, 5.690±0.080), and the expressions of DR6 and p65 were decreased (1.53±0.09, 0.41±0.04). Conclusion:miR-106a can reduce the inflammatory changes and the degeneration of chondrocytes in osteoarthritis rats.

Optimization of expression conditions and determination the proteolytic activity of codon-optimized SARS-CoV-2 main protease in Escherichia coli / 生物工程学报

The main protease (Mpro) of SARS-CoV-2 is a highly conserved and mutation-resistant coronaviral enzyme, which plays a pivotal role in viral replication, making it an ideal target for the development of novel broad-spectrum anti-coronaviral drugs. In this study, a codon-optimized Mpro gene was cloned into pET-21a and pET-28a expression vectors. The recombinant plasmids were transformed into E. coli Rosetta(DE3) competent cells and the expression conditions were optimized. The highly expressed recombinant proteins, Mpro and Mpro-28, were purified by HisTrapTM chelating column and its proteolytic activity was determined by a fluorescence resonance energy transfer (FRET) assay. The FRET assay showed that Mpro exhibits a desirable proteolytic activity (25 000 U/mg), with Km and kcat values of 11.68 μmol/L and 0.037/s, respectively. The specific activity of Mpro is 25 times that of Mpro-28, a fusion protein carrying a polyhistidine tag at the N and C termini, indicating additional residues at the N terminus of Mpro, but not at the C terminus, are detrimental to its proteolytic activity. The preparation of active SARS-CoV-2 Mpro through codon-optimization strategy might facilitate the development of the rapid screening assays for the discovery of broad-spectrum anti-coronaviral drugs targeting Mpro.

Application of Debriefing in clinical medicine: a bibliometric analysis of Debriefing related research papers based on data in Web of Science / 中华医学教育探索杂志

Objective:To make bibliometric analysis of Debriefing related research papers, and to lay a foundation for further research.Methods:A bibliometric analysis was conducted on the distribution of years, nations, institutions, the situation of citations, and main research contents of Debriefing related researches.Results:Analysis of the distribution of the publication years of Debriefing related literature indicated that since the relevant literature of Debriefing had been reported in 1979, its publication volume has been increasing year by year, mainly published by the United States, the United Kingdom, Canada and other countries. In addition, most journals that publish more literature on Debriefing are related to medical education, and most of the highly cited studies focus on relevant research theories based on Debriefing. Debriefing may be mainly applied in clinical teaching, nursing students' simulation teaching and training and its clinical application, among which nursing, psychiatry, emergency, internal medicine, pediatrics, surgery, anesthesiology, and other departments are common.Conclusion:Through the bibliometric analysis of Debriefing related research papers, this paper has a preliminary understanding of the status quo and hotspots of this field, which lays a good foundation for the subsequent research.

Mining Polo-Box domain of Polo-like kinase 1 as a new therapeutic target for cancer / 生物工程学报

Polo-like kinase 1 (Plk1) is widely regarded as one of the most promising targets for cancer therapy due to its essential role in cell division and tumor cell survival. At present, most Plk1 inhibitors have been developed based on kinase domain, some of which are in clinical trial. However, inhibitors targeting kinase domain face off-target effect and drug resistance owing to the conserved nature and the frequent mutations in the ATP-binding pocket. In addition to a highly conserved kinase domain, Plk1 also contains a unique Polo-Box domain (PBD), which is essential for Plk1's subcellular localization and mitotic functions. Inhibitors targeting Plk1 PBD show stronger selectivity and less drug resistance for cancer therapy. Therefore, Plk1 PBD is an attractive target for the development of anti-cancer agents. In this review, we will summarize the up-to date drug discovery for targeting Plk1 PBD, including the molecular structure and cellular functions of Plk1 PBD. Small-molecule inhibitors targeting Plk1 PBD not only provide an opportunity to specifically inhibit Plk1 activity for cancer treatment, but also unveil novel biological basis regarding the molecular recognition of Plk1 and its substrates.

Mid-term results of endovascular treatment for the transplanted renal arterial stenosis / 中华普通外科杂志

Objective To investigate the safety and mid-term efficacy of percutaneous endovascular angioplasty(PTA) and stent implantation for transplantation renal arterial stenosis (TRAS).Methods Retrospective analysis was performed on 18 patients with TRAS admitted to of department vascular surgery,PLA General Hospital from Jan 2011 to Dec 2018.Results PTA and stent implantation were performed in all 18 patients via ipsilateral or contralateral femoral artery including 4 cases of PTA alone,8 cases of PTA plus stent,6 cases of stent implanted directly.Three of the 18 patients underwent ipsilateral femoral artery catheterization and 15 underwent contralaterally.A total of 14 stents were implanted,all of were balloon expanding stents,of which 2 were drug-coated stents,and the technical success rate was 100％.The average dosage of contrast agent was 64 ml,the stenosis rate of renal artery before interventional treatment was 50％-99％,and that after interventional treatment reduced to 10％-30％.The systolic blood pressure decreased from (157.2 ± 43.0) mmHg preoperatively to (129.8 ± 8.6) mmHg postoperatively.The SCr level decreased from (258.8 ± 214.7) μ mol/L to (176.3 ± 101.1) μmol/L.Preoperative urea nitrogen decreased from (15.7 ± 1.6) mmol/L to (10.6 ± 1.1) mmol/L postoperatively (P ＜ 0.05).Mean postoperative follow-up time was 42.4 months (3-93 months).17 cases were cured,1 case was ineffective,1 case suffered restenosis after 30 days,and was given remedy PTA plus stenting.Conclusions TRAS is a vascular factor leading to grafted renal failure,the endovascular treatment of TRAS is safe,effective and has good mid-term result.

The midterm result of drug coated balloon with paclitaxel for the treatment of TASC C/D superficial femora-popliteal artery disease / 中华普通外科杂志

Objective@#To evaluate the safety and efficacy of the drug coated balloon(DCB) for complex TASC C/D superficial femora-popliteal artery diseases.@*Methods@#Patency, target lesion revascularization (TLR) rate, clinical improvement and safety endpoints of femora-popliteal lesions in 68 patients from PLA General Hospital treated with DCB were retrospectively analyzed from June 2016 to June 2018. The mean age of the patients were (72.7±13.2) years old. Rutherford categories were from 2 to 5, and ABI baseline were 0.56±0.22.@*Results@#There were 76 limbs treated by DCB in total in this study. Mean lesion length was (26.7±15.3) cm. 73.6% of lesions were totally occluded, 26.4% were of stenosis and 61.8% were highly calcified. Stent implantation was performed in 36.8% cases. Kaplan Meier estimates of primary patency were 74.2%±7.6% and 67.7%±6.4% at 1 and 2 years, respectively, whereas freedom from TLR was 81.4%±5.1% and 73.6%±5.4%. ABI were 0.83±0.16 at 1 year, and 0.79±0.24 at 2 years. Major amputation rate was 2.9% and mortality rate was 2.9% and 4.4% at 1 year and 2 years respectively. Diabetes, highly calcification, renal insufficiency and re-stenotic lesions were identified as predictors of restenosis.@*Conclusions@#DCB are safe and effective in delaying restenosis in complex TASC C/D superficial femora-popliteal artery disease as found by midterm follow-up.

Electric acupuncture plus fire acupuncture improves facial nerve function and anxiety state in patients with obstinate facial palsy / 中华全科医师杂志

One hundred and ten patients with obstinate facial palsy were randomly divided into study group and control group with 55 cases in each group. Both groups received electropuncture on face acupoints five times a week, the study group received additional acupuncture on Zhongzhen six acupoints, subsequently fire needles were used for Yangbai (GB 14), Quanliao (SI 18), Dicang (ST 4) and Jiachengjiang (Extra) acupoints twice a week. And the treatment courses lasted for four weeks. The House-Brackmann (HB) scale and Self-Rating Anxiety Scale (SAS) were used for evaluation of facial nerve function and anxiety status before and after treatment, respectively. One hundred and four cases completed the treatment in the two groups. Before treatment there were no significant differences in HB and SAS scores between study and control groups (Z=-0.271, P=0.79; Z=-1.689, P=0.09). After 4 weeks of treatment the HB and SAS scores in both group were significantly improved (Z=-5.652, Z=-5.038, all P<0.01; Z=-6.050, Z=-5.332, all P<0.01); however, there were significant differences in HB scale and SAS scores after treatment between two groups (Z=-2.066, Z=-2.884, all P<0.05). The electric acupuncture on face acupoints plus Zhongzhen six acupoints combined with fire acupuncture can effectively improve facial nerve function and anxiety state in patients with obstinate facial palsy.

Electric acupuncture plus fire acupuncture improves facial nerve function and anxiety state in patients with obstinate facial palsy / 中华全科医师杂志

One hundred and ten patients with obstinate facial palsy were randomly divided into study group and control group with 55 cases in each group.Both groups received electropuncture on face acupoints five times a week,the study group received additional acupuncture on Zhongzhen six acupoints,subsequently fire needles were used for Yangbai (GB 14),Quanliao (SI 18),Dicang (ST 4) and Jiachengjiang (Extra) acupoints twice a week.And the treatment courses lasted for four weeks.The House-Brackmann (HB) scale and Self-Rating Anxiety Scale (SAS) were used for evaluation of facial nerve function and anxiety status before and after treatment,respectively.One hundred and four cases completed the treatment in the two groups.Before treatment there were no significant differences in HB and SAS scores between study and control groups (Z=-0.271,P=0.79;Z=-1.689,P=0.09).After 4 weeks of treatment the HB and SAS scores in both group were significantly improved (Z=-5.652,Z=-5.038,all P＜0.01;Z=-6.050,Z=-5.332,all P＜0.01);however,there were significant differences in HB scale and SAS scores after treatment between two groups (Z=-2.066,Z=-2.884,all P＜0.05).The electric acupuncture on face acupoints plus Zhongzhen six acupoints combined with fire acupuncture can effectively improve facial nerve function and anxiety state in patients with obstinate facial palsy.

Comparative analysis of the clinical characteristics of elderly onset rheumatoid arthritis and elderly onset rheumatoid arthritis with osteoarthritis / 中华风湿病学杂志

Objective To compare the clinical features,ultrasonic imaging manifestations and therapeutic evaluations between elderly onset rheumatoid arthritis (EORA) and EORA with osteoarthritis (OA).Methods Eighty-eight patients with rheumatoid arthritis were divided into two groups:group EORA (n=36)and group EORA+OA (n=52).The onset age of all patients was 60 years or older.General conditions,joint involvement distribution,ultrasonic manifestations and disease activity scores (DAS28-3) of patients in the two groups were analyzed.The x2 test/Fisher's exact probability test and the Student's t test/Mann-Whitney U test were used to analyze data.Results There was no significant difference in the proportion of male and female patients and erythrocyte sedimentation rate (ESR) between the two groups (P＞0.05).The onset age of patients in group EORA+OA [(68±4) years old] was higher than that in group EORA [(65±4) years old],and the difference was statistically significant (t=-3.465,P=0.001).Duration of the disease and body mass index in group EORA+OA were significantly higher respectively than those in group EORA.Joint involvement in the two groups was mainly found in shoulder,wrist,Metacarpophalangeal joint (MCP)2,MCP3,proximal interphalangeal joint (PIP)2,PIP3,PIP4,and knee joint (34.7％-86.5％).The percentage of MCP2 [36.5％(38/104),70.8％ (51/72);x2 =20.02,P ＜0.01],MCP3 [33.7％ (35/100),59.7％ (43/72);x2 =11.72,P =0.001],MCP4 [4.8％ (5/100),22.2％ (16/72);x2 =12.28,P＜0.01],PIP2 [69.2％ (72/104),83.3％ (60/72);x2 =4.51,P=0.034]and PIP3 [53.8％(56/100),70.8％(51/72);x2=5.15,P=0.023] in the EORA+OA group was lower while the percentage of MCP1,DIP 2,DIP3,DIP4 and knee joints were higher than that in the EORA group (P＜0.05).In group EORA+OA,the synovial thickness of the wrist joints [(4.2±0.5) mm] and knee joints [(7.7±0.8) mm]were significantly thicker than those in group EORA [(3.2±0.9) mm;(6.3±0.8) mm,t=-5.82,P＜0.01;t=-7.22,P＜0.01];The proportion (70.0％) of level 2 and 3 of patients' wrist joint synovium pannus blood flow and knee joint synovium pannus in group EORA+OA were increased than group EORA (51.9％;52.3％),the difference between the two groups was statistically significant (x2=4.64,P=0.031;x2=4.43,P=0.035).There was no significant difference in DAS28-3 scores between the two groups before patients received treatment.After 2 weeks and 12 weeks of glucocorticoid treatment,DAS28-3 scores in group EORA [3.62 (2.88,4.03);2.35 (2.26,2.62) points] were significantly lower than group EORA+OA [5.01(4.68,5.26);3.38(2.28,3.83) points](Z=-7.766,P＜0.01;Z=-3.461,P＜0.01).Conclusion Compared with patients of EORA alone,patients of EORA with OA have more obvious joint symptoms,MCP1,DIP and knee joint are susceptible to the coinvolvement among them,longer duration of disease,and were prone to synovial hyperplasia and pannus flow formation.The therapeutic effects of glucocorticoid on joint inflammation in patients of EORA alone are superior to those patients of EORA with OA.

The midterm result of drug coated balloon with paclitaxel for the treatment of TASC C/D superficial femora-popliteal artery disease / 中华普通外科杂志

Objective To evaluate the safety and efficacy of the drug coated balloon (DCB) for complex TASC C/D superficial femora-popliteal artery diseases.Methods Patency,target lesion revascularization (TLR) rate,clinical improvement and safety endpoints of femora-popliteal lesions in 68 patients from PLA General Hospital treated with DCB were retrospectively analyzed from June 2016 to June 2018.The mean age of the patients were (72.7 ± 13.2) years old.Rutherford categories were from 2 to 5,and ABI baseline were 0.56 ± 0.22.Results There were 76 limbs treated by DCB in total in this study.Mean lesion length was (26.7 ± 15.3) cm.73.6％ of lesions were totally occluded,26.4％ were of stenosis and 61.8％ were highly calcified.Stent implantation was performed in 36.8％ cases.Kaplan Meier estimates of primary patency were 74.2％ ± 7.6％ and 67.7％ ± 6.4％ at 1 and 2 years,respectively,whereas freedom from TLR was 81.4％ ±5.1％ and 73.6％ ±5.4％.ABI were 0.83 ±0.16 at 1 year,and 0.79 ±0.24 at 2 years.Major amputation rate was 2.9％ and mortality rate was 2.9％ and 4.4％ at 1 year and 2 years respectively.Diabetes,highly calcification,renal insufficiency and re-stenotic lesions were identified as predictors of restenosis.Conclusions DCB are safe and effective in delaying restenosis in complex TASC C/D superficial femora-popliteal artery disease as found by midterm follow-up.

Screening of pathogenic genes in a Chinese familial dilated cardiomyopathy pedigree from Inner Mongolia / 中华心血管病杂志

Objective@#Screen the pathogenic genes of a pedigree with clinical manifestation of familial dilated cardiomyopathy in Inner Mongolia.@*Methods@#A total of 3 patients with dilated cardiomyopathy and 20 family members from the same family were examined in Ordos Central Hospital in Inner Mongolia from October, 2003 to August, 2017. Data on medical history, physical examinations, electrocardiograms, and echocardiography were obtained. 5 ml peripheral blood was sampled for per person. Chip Capture Sequencing technology was used to capture all the exons and splice sites of the genes that associated with hereditary cardiomyopathy and hereditary arrhythmia. The mutations in these genes were detected by high-throughput sequencing. All suspected pathogenic loci identified by high-throughput sequencing were verified by Sanger sequencing used for mutation detection. One hundred and fifty gender, age and race matched healthy people were included as the control group.@*Results@#Pathogenic gene variations were detected in 3 symptomatic family members and 1 carrier from the pedigree. Five pathogenic gene variations were identified in the proband (Ⅱ1), a pSer236Gly and a pArg215Cys variation in the MYBPC3 gene, a pGln90Arg variation in the DSP gene, and pAsn2912Asp and pGlu2910Val variation in the DMD gene. One pathogenic variation was detected in Ⅲ3, which was a pArg215Cys variation in the MYBPC3 gene. Two pathogenic variations were detected in Ⅲ7, a pSer236Gly variation in the MYBPC3 gene and a pGln90Arg variation in the DSP gene. Two pathogenic variations were detected in the Ⅳ7, a pSer236Gly variation in the MYBPC3 gene and a pGln90Arg variation in the DSP gene. No gene variation loci were detected in the other family members and the control group.@*Conclusion@#MYBPC3 gene, DSP gene and DMD gene variations are present in the familial dilated cardiomyopathy pedigree from Inner Mongolia, and these variations may be related with familial dilated cardiomyopathy.

Comparative analysis of the clinical characteristics of elderly onset rheumatoid arthritis and elderly onset rheumatoid arthritis with osteoarthritis / 中华风湿病学杂志

Objective@#To compare the clinical features, ultrasonic imaging manifestations and therapeutic evaluations between elderly onset rheumatoid arthritis (EORA) and EORA with osteoarthritis (OA).@*Methods@#Eighty-eight patients with rheumatoid arthritis were divided into two groups: group EORA (n=36) and group EORA+OA (n=52). The onset age of all patients was 60 years or older. General conditions, joint involvement distribution, ultrasonic manifestations and disease activity scores (DAS28-3) of patients in the two groups were analyzed. The χ2 test/Fisher's exact probability test and the Student's t test/Mann-Whitney U test were used to analyze data.@*Results@#There was no significant difference in the proportion of male and female patients and erythrocyte sedimentation rate (ESR) between the two groups (P>0.05). The onset age of patients in group EORA+OA [(68±4) years old] was higher than that in group EORA [(65±4) years old], and the difference was statistically significant (t=-3.465, P=0.001). Duration of the disease and body mass index in group EORA+OA were significantly higher respectively than those in group EORA. Joint involvement in the two groups was mainly found in shoulder, wrist, Metacarpophalangeal joint (MCP)2, MCP3, proximal inter-phalangeal joint (PIP)2, PIP3, PIP4, and knee joint (34.7%-86.5%). The percentage of MCP2[36.5%(38/104), 70.8%(51/72); χ2=20.02, P<0.01], MCP3[33.7%(35/104), 59.7%(43/72); χ2=11.72, P=0.001], MCP4[4.8%(5/104), 22.2%(16/72); χ2=12.28, P<0.01], PIP2[69.2%(72/104), 83.3%(60/72); χ2=4.51, P=0.034] and PIP3[53.8%(56/104), 70.8%(51/72); χ2=5.15, P=0.023] in the EORA+OA group was lower while the percentage of MCP1, DIP 2, DIP3, DIP4 and knee joints were higher than that in the EORA group (P<0.05). In group EORA+OA, the synovial thickness of the wrist joints [(4.2±0.5) mm] and knee joints [(7.7±0.8) mm] were significantly thicker than those in group EORA [(3.2±0.9) mm; (6.3±0.8) mm, t=-5.82, P<0.01; t=-7.22, P<0.01]; The proportion (70.0%) of level 2 and 3 of patients' wrist joint synovium pannus blood flow and knee joint synovium pannus in group EORA+OA were increased than group EORA (51.9%; 52.3%), the difference between the two groups was statistically significant (χ2=4.64, P=0.031; χ2=4.43, P=0.035). There was no significant difference in DAS28-3 scores between the two groups before patients received treatment. After 2 weeks and 12 weeks of glucocorticoid treatment, DAS28-3 scores in group EORA [3.62(2.88, 4.03); 2.35(2.26, 2.62) points] were significantly lower than group EORA+OA [5.01(4.68, 5.26); 3.38(2.28, 3.83) points] (Z=-7.766, P<0.01; Z=-3.461, P<0.01).@*Conclusion@#Compared with patients of EORA alone, patients of EORA with OA have more obvious joint symptoms, MCP1, DIP and knee joint are susceptible to the co-involvement among them, longer duration of disease, and were prone to synovial hyperplasia and pannus flow formation. The therapeutic effects of glucocorticoid on joint inflammation in patients of EORA alone are superior to those patients of EORA with OA.

Development of an ELISA-based high throughput screening method for novel anticancer agents targeting β-catenin/Lef1 interaction / 生物工程学报

To develop an enzyme-linked immunosorbent assay (ELISA)-based high throughput screening (HTS) method for β-catenin/Lef1 interaction antagonists screening, Escherichia coli Rosetta (DE3) competent cells were transformed with β-catenin-pET-30a(+) plasmid. β-catenin protein was expressed after induction and purified using affinity chromatography. The biological activity of purified β-catenin was further analyzed by GST Pulldown assay. The β-catenin/GST-Lef1 binding model was established using ELISA principle, and the ELISA-based HTS method was further optimized through determination of an optimal coated concentration of GST-Lef1 and working concentration of β-catenin. The results showed that β-catenin protein was successfully expressed and purified. The GST Pulldown assay demonstrated a perfect biological activity for purified β-catenin. Subsequently, the ELISA-based HTS method was performed using 10 μg/mL GST-Lef1 and 6 μg/mL β-catenin, with the Z factor of 0.76. Taken together, we have successfully developed a simple, robust and reliable ELISA-based HTS method for screening of novel Wnt inhibitors targeting β-catenin/Lef1 interaction.

Experimental study of serum SM22 in the diagnosis of acute intestinal ischemia / 中华普通外科杂志

Objective To evaluate smooth muscle protein of 22 kDa (SM22) in the diagnosis of acute intestinal ischemia.Methods 96 healthy adult SD rats were evenly divided into experimental group and control group,with each group subdivided into 6 subgroups,subject respectively to superior mesenteric artery ligation or sham operation.The venous blood samples were extracted from each group rats' right heart atO.5,1,2,4,8,12 h after the operation,for SM22 testing and small intestines tissues for direct immunofluorescence staining of SM22.Results The serum SM22 concentration reached a peak at 4 h (265 ± 15) mg/L,then gradually decreased (P ＜ 0.05).The I-FABP was mainly expressed in the epithelium of intestinal mucosa.During the 4 hours of intestinal ischemia,The number of SM22 positive particles did not change.After 4 hours,the number of SM22 positive granules had gradually decreased compared with the control group (all P ＜ 0.05).Conclusion SM22 mainly exists in the smooth muscle of intestinal,during the ischemic necrosis of the intestinal muscle layer SM22 leaks into blood stream,resulting in high serum levels of SM22 facilitating early diagnosis of acute intestinal ischemia.

Bacterial expression, preparation and identification of polyclonal antibody against Escherichia coli FtsZ / 生物工程学报

To prepare polyclonal antibody (PcAb) against Escherichia coli filamentous thermosensitive protein Z (Ec-FtsZ), the artificially synthesized gene fragment coding Ec-FtsZ was subcloned into pET-22b(+) plasmid, and Ec-FtsZ protein was expressed in E. coli BL21(DE3) cell under an optimal bacterial expression condition. Then Ec-FtsZ protein was purified by HisTrap affinity chromatography, and the GTPase (Guanosine triphosphatase) activity of purified Ec-FtsZ protein was further analyzed by malachite green assay. Subsequently, the purified Ec-FtsZ protein was used to immunize rat subcutaneously for preparation of anti-Ec-FtsZ PcAb. The results of enzyme-linked immunosorbent assay (ELISA), Western blotting analysis and immunofluorescence assay showed that the titer of PcAb was 1:256 000, and PcAb exhibited a perfect antigenic specificity against purified and endogenous Ec-FtsZ protein. All these data indicated that the anti-Ec-FtsZ PcAb is successfully prepared, which can be used for further cellular function study and biochemical analysis of Ec-FtsZ protein in vivo.

Applied research of archived medical record management in medical quality management / 中华医院管理杂志

Objective@#To summarize the impacts of the archived medical record value fund as established by the hospital, on the medical quality management, and propose improvement strategies.@*Methods@#Retrospective analysis was made on 1 149 medical record scoring sheets, and the rate of connotations deducting scores in 197 of such sheets were compared. A statistical analysis was made on the archived medical records, regarding the rate of grade A medical records, defects composition of grade B medical records, the rate of connotation deducting scores, and the ability of the quality control experts (error correction ratio).@*Results@#The archived medical record value fund has pushed up significantly the quality of medical records. For example, the rate of grade A medical records rose step by step from 89% to 95%; the rate of grade B medical records with single-item vetoes fell sharply, a difference of statistical significance (P<0.05); the connotation quality management of medical records was constantly improving, with the rate of connotation deducting scores gradually elevating (t=-4.409, P<0.05); the proportion of experts with strong error correction ability (error correction ratio≥80%) was elevated from 32.2% to 50.9%, a difference of statistical significance (P<0.05).@*Conclusions@#Archived medical record management can encourage medical quality management. The quality of medical records, especially the connotation quality of medical records has been improved. The ability of some quality control experts was also enhanced. The quality management system at hospital and department levels has made major progress.