Objective:To study the expression of kringle 1-3 domain fragment of human plasmihogen in E.coli and the anti-tumor activity of its product.Methods:The K1-3 domain fragment was cloned in expression vector pBV220,the resulted recombinant plasmid pBV-K13 was transformed into E.coli DH5? and its product was purified and assayed its bioactivity.Results:K1-3 domain fragment was expressed in(E.coli) DH5?.The results showed the expressed product covered 20% of the total bacterial protein on SDS-PAGE and the Western blot analysis showed that the product had immunological specificity with the antiserum of human plasminogen and inhibits the growth of chorioallantoic membrane(CAM) angiogenesis and mouse B16 melanoma.Conclusion:Human plasminogen K1-3 domain fragment was expressed in E.coli;the expressed product has anti-angiogenesis and anti-tumor activity.

AIM: Constructing plasmid that expresses human plasminogen kringle 5 gene to analyze the gene expression in E.coli. METHODS: The gene of human plasminogen kringle 5 was inserted into plasmid PBV220 EcoRI site by gene manipulation techniques and was transformed to E.coli TGI. The gene expression was observed by SDS-PAGE. RESULTS: Expression vector PBVK5 was constructed, and human plasmingen kringle 5 gene product was obtained at 42℃ induction. CONCLUSLON: Expression product of human plasminogen kringle 5 gene was soluble form of proteins, and the expression amount was 9.8% in E.coli TGI total proteins.

Objective To investigate the value of baseline levels of aldosterone(ALD)in predicting the cardiovascular events and in-stent restenosis12 months after coronary stent implantation. Methods 268 patients with coronary heart disease admitted to the department of cardiology from January 2014 to July 2016 were selected (96 cases of stable angina pectoris and 172 cases of acute coronary syndrome).The ALD level in the preprocedural serum was detected before coronary stent implantation. According to the level of ALD,the patients were divided into two groups:ALD<130 pg/mL(n=127)and ALD≥130 pg/mL(n=141).The cardiovascular events(angina pectoris,myocardial infarction,death)and in-stent restenosis were observed and recorded 12 months after the procedures. Logistic regression analysis was done to analyze the independent risk factors of ISR. Results The baseline levels of ALD in the patients with acute coronary syndrome were significantly higher than those with stable angina pectoris[(160 ± 58)pg/mL vs.(118 ± 46)pg/mL,P < 0.05]. The cardiovascular events and those in in-stent restenosis in the patients with high baseline levels of ALD were significantly higher than those with normal baseline levels of ALD 12 months too(24.1% vs 3.1%;P<0.05;16% vs 7.4%;P<0.05).Logistic regression analysis showed that serum ALD levels were the strongest predictors of ISR,with the odds ratio of 2.56 per 10 pg/mL. Conclusions Preprocedural ALD level is a predictor for 12 months outcome of in-stent restenosis for the patents undergoing coronary stent implantation. It indicates that the complications and clinical restenosis in 12 months are markedly influenced by the activation of the rennin angiotensin aldostenone system.

The claustrum located between the striatum and cortex is composed of glutamate neurons and gamma-aminobutyric neurons. There are extensive and mutually projected nerve fiber connections between cerebral cortex and parietal nucleus. The claustrum integrates sensory information and regulates the formation of consciousness through various ways, playing an important role in sensory processing and cognitive functions. In this paper, the neuronal characteristics, fiber projection, physiological characteristics and consciousness integration function of the claustrum are reviewed to provide references for further understanding physiological and pathophysiological effects of the pterygoid nucleus.

Objective:To explore the long-term efficacy of a modified unilateral cutaneous ureterostomy in bladder cancer patients receiving radical cystectomy.Methods:The medical data of 104 bladder cancer patients who underwent ureterostomy in our hospital from Janurary 2013 to December 2020 were retrospectively analyzed. The patients were divided into unilateral and bilateral group. The unilateral group contained 66 cases, with 53 males and 13 females, average age (71.8±9.8) years, body mass index (BMI)(23.3±3.2)kg/m 2. The bilateral group contained 38 cases, with 33 males and 5 females, average age (75.1±10.8) years; BMI (22.7±3.0)kg/m 2. There was no significant difference in the above characteristics between the two groups ( P>0.05). The pathology, survival status, long-term complications between the two groups were compared. Quality of life was assessed during follow-up using the European Core Questionnaire for Quality of Life in Cancer Patients (EORTC QLQ-C30). Results:The unilateral group contained 46(69.7%) muscle invasive bladder cancer (MIBC) cases, 15 (22.7%) cases with lymph node metastasis, 7 (10.6%) cases with distant metastasis. The bilateral group contained 24(63.2%) muscle invasive bladder cancer(MIBC) cases, 6 (15.8%) cases with lymph node metastasis, 2 (5.3%) cases with distant metastasis. There was no significant difference in disease specific survival between the two groups ( P>0.05). During the follow-up, the incidence of overall complication rate in the unilateral group was significantly lower than that in the bilateral group [43.9% (29/66) vs. 63.2% (24/38), P<0.001]. The incidence of pyelonephritis in unilateral group was significantly lower than that in the bilateral group [16.6%(11/66) vs. 42.1%(16/38), P=0.006]. There was no statistical significance in terms of quality of life before operation in the two groups. After operation, both physical function score[(54.9±7.1) vs.(49.2±6.7)] and emotional function score [(63.1±6.4) vs.(59.9±6.7)] in unilateral group were higher than that in bilateral group ( P<0.05). Conclusions:The modified unilateral cutaneous ureterostomy could achieve relatively low complication rate, and improve the quality of life to some extent compared with bilateral ureterostomy.

Objective To investigate the efficacy and safety of Nocardiarubra cell wall skeleton (N-CWS) bladder irrigation in prevention of recurrence after transurethral resection for the treatment of non-muscle invasive bladder cancer (NMIBC).Methods The clinical data of patients with NMIBC treated by N-CWS and epirubicin collected between October 2013 and November 2018 at the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed.All patients underwent TURBT.Among the 118 NMIBC patients,the average age was (65.1 ± 1 1.9) years,and the sex ratio (male/female) was 1.9∶1 (77/41).Patients were divided into two group:N-CWS group (n =55) and epirubicin group (n =63) according to different instillation regimens.N-CWS was given as an instillation of 800 μg in 50 ml of saline and maintained in the bladder for 2 h in the N-CWS group.Epirubicin was given as an instillation of 50 mg in 50 ml of saline and maintained in the bladder for 1 h in the epirubicin group.In the N-CWS group,mean agewas (64.9 ± 12.1) years and 37 (67.3％) were male.Multiple tumors were present in 17 (69.1％) patients.Tumor size was ≤3 cm in 49(89.1％) and 7(12.7％) had a history of NMIBC.Stage was Ta and T1 in 36(65.5％) and 19(34.5％),respectively.Grade 1,Grade 2 and Grade 3 were the primary grades in 38(69.1％),13(23.6％) and 4(7.3％),respectively.Low risk,intermediate risk and high risk were present in 14 patients(25.5％),16 (29.1％) and 25 (45.5％),respectively.In the epirubicin group,mean age was (65.3 ± 11.2) years and 40(63.5％)were male.Multiple tumors were present in 19(30.2％) patients.Tumor size was ≤3 cm in 56(88.9％) and 11 (17.5％) had a history of NMIBC.Stage was Ta and T1 in 37(58.7％) and 26 (41.3％),respectively.Grade 1,Grade 2 and Grade 3 were the primary grades in 44(69.8％),12(19.0％)and 7(11.1％),respectively.Low risk,intermediate risk and high risk were present in 13 (20.6％),19 (30.2％) and 31 (49.2％),respectively.The tumor recurrence,progression and adverse reactions after Intravesical Instillation in both groups were followed up and recorded.No significant differences were found between the two groups.Results A total of 118 patients were followed up.Mean follow-up time was (33.7 ± 5.4) months.25.5％ (14/55) in the N-CWS group vs.42.8％ (27/63) in the epirubicin group had recurrence after 5 years (x2 =3.922,P =0.048).The five-year RFS was higher in the N-CWS group than in the epirubicin group (74.2％ vs.56.5％,P =0.044).No significant difference was found in the progression rate between the two groups(5.5％ vs.7.9％,P =0.867).The incidences of adverse events in the two groups were 16.4％ (9/55) and 19.0％ (12/63),respectively.The N-CWS group had significantly fewer cases with urinary frequency and dysuria than the epirubicin group.No significant differences were found in other side effects.Conclusions Intravesical instillation of N-CWS after NMIBC TURBT was found to be a promising procedure to prevent recurrence and prolong the recurrence-free survival with less side effects.

Objective To summary the pathological and clinical characteristics,treatments and prognosis of malignant renal perivascular epithelioid cell tumor (PEComa).Methods Between August 2013 and June 2017,8 cases of malignant renal PEComa were analysed respectively.Of all the patients,there were 4 males and 4 females,aged 27-65 years with the average of 46.3 years old.Three cases were detected in routine examination occasionally,three cases complained of intermittent back pain with fever,one presented with swollen and painful right hand and right foot,and one case presented with hematuria.CT or MRI examinations indicated the malignant lesions before the surgery.Clinical staging was performed with 2 cases of T1N0M0,1 case of T1 N0M1,2 cases of T2N0M0,1 case of T3M0M0,and 2 cases of T4N0M1.Three cases underwent radical nephrectomy,1 case underwent radical nephrectomy plus renal artery embolization,2 cases underwent partial nephrectomy,and 2 cases underwent nephrectomy plus inferior vena cava tumor thrombectomy.One case was treated with ifosfamide plus epirubicin after operation due to multiple distant metastases and 1 case was treated with oxaliplatin before the sugery because of excessive tumor diameter.None of the rest received any adjuvant therapy.Results Postoperative histopathological examination showed multiple nodules in reddish gray and yellow color,with soft texture and partial incomplete capsule.Microscopically,there were large atypical cell components,some of which were spindle shaped,with disordered cell arrangement,some of which were associated with a large amount of necrotic tissue,and abundant light eosinophil cytoplasm.Tissue components can be found thick-wall blood vessels,smooth muscle-like cells,and adipose tissue.Immunohistochemistry showed 4 cases were positive of HMB-45,4 of Melan-A,7 of SAM.Seven cases were negative of CK,6 of S-100 75％ (6/8) and Ki67 (10％-70％).Two cases died of multiple metastases (lung,bone,liver),1 case survived with tumor recurrence,with a follow-up from 14 to 60 months (mean 29 months).Conclusions Primary malignant renal PEComa is rare in practice with relative large diameter.The diagnosis depends on pathological findings,and radical nephrectomy is the first choice of treatment.It is easy to recur and metastasize after the operation.

【Objective】 To explore the prognosis of critically ill patients with coagulation dysfunction using thrombelastogram(TEG) and coagulation four items combined with APACHEⅡ score. 【Methods】 From March 2017 to March 2020, 287 critically ill patients with coagulation dysfunction in our hospital were selected as the study group, and 303 patients with normal coagulation function during the same period were set as the control. The study group was divided into low-risk group(group A), intermediate-risk group(group B) and high-risk group (group C) based on the APACHEⅡ score, and into survival group and death group according to the prognosis. The difference of TEG, coagulation four items, and APACHEⅡ scores between the two groups were analyzed. The correlation and difference between TEG, coagulation four items and APACHE II score in the study group were analyzed. The ROC curve was drawn to analyze the prognostic predictive value of research indicators. 【Results】 Blood coagulation function related indicators in the study group fluctuated significantly: in comparison to the control, the CI value, MA value, and α angle were smaller, while the K time and R time were longer; among the coagulation four items, PT, APTT and TT were higher; Fg level was lower, and the APACHE Ⅱ score was higher(P<0.05). There were significant differences in relevant test index levels among patients with different degree of illness (APACHE Ⅱ score) in the study group. With the aggravation of the disease, the CI value, MA value, α angle, and TT continued to decrease, while K time, R time, APTT and PT showed a continuous increase trend (P<0.05). However, with the intensification of coagulation dysfunction, no significant increase or decrease was noticed in Fg(P>0.05). There were significant differences between the TEG and coagulation function related index levels in patients with different prognosis. Compared with the survivals, the CI value, MA value and α angle of the dead group were smaller, while the K time and R time were longer; and among the coagulation four items, PT, APTT, and TT were higher, the Fg level was lower, and the APACHEⅡ score was higher (P<0.05). The ROC curve showed that the corresponding AUC values of PT, APTT and INR were 0.701, 0.693 and 0.702, respectively, (P<0.05). The difference was statistically significant and had predictive value for death, but the accuracy was moderate.The combined indicators showed that the AUC values corresponding to APACHE Ⅱ score, P1, P2, P3, P4, P5, and P6 were 0.899, 0.751, 0.657, 0.759, 0.921, 0.921and 0.942, respectively, and the difference was statistically significant(P<0.05). The combined indicators have predictive value for death, except for P2<0.7, the rest were between 0.7~1.0, and the accuracy was P6>P4\\P5>APACHE Ⅱ score>P1>P2. 【Conclusion】 TEG, coagulation four items, and APACHE Ⅱ score can be used to assess the severity of patients with severe coagulation dysfunction. and the combined application of the 3 indicators are of high value in predicting the prognosis of such patients, and can provide reference for clinical formulation or adjustment of intervention programs to correct coagulation dysfunction.

Glutamate receptor-like (GLR) is an important class of Ca²⁺ channel proteins, playing important roles in plant growth and development as well as in response to biotic and abiotic stresses. In this paper, we performed genome-wide identification of banana GLR gene family based on banana genomic data. Moreover, we analyzed the basic physicochemical properties, gene structure, conserved motifs, promoter cis-acting elements, evolutionary relationships, and used real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) to verify the expression patterns of some GLR family members under low temperature of 4 ℃ and different hormone treatments. The results showed that there were 19 MaGLR family members in Musa acuminata, 16 MbGLR family members in Musa balbisiana and 14 MiGLR family members in Musa itinerans. Most of the members were stable proteins and had signal peptides, all of them had 3-6 transmembrane structures. Prediction of subcellular localization indicated that all of them were localized on the plasma membrane and irregularly distributed on the chromosome. Phylogenetic analysis revealed that banana GLRs could be divided into 3 subclades. The results of promoter cis-acting elements and transcription factor binding site prediction showed that there were multiple hormone- and stress-related response elements and 18 TFBS in banana GLR. RT-qPCR analysis showed that MaGLR1.1 and MaGLR3.5 responded positively to low temperature stress and were significantly expressed in abscisic acid/methyl jasmonate treatments. In conclusion, the results of this study suggest that GLR, a highly conserved family of ion channels, may play an important role in the growth and development process and stress resistance of banana.
Musa/metabolism* ; Phylogeny ; Abscisic Acid/metabolism* ; Temperature ; Stress, Physiological/genetics* ; Hormones/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Gene Expression Profiling

Tripterygium wilfordii is a valuable medicinal plant rich in biologically active diterpenoids, but there are few studies on the origins of these diterpenoids in its secondary metabolism. Here, we identified three regions containing tandemly duplicated diterpene synthase genes on chromosomes (Chr) 17 and 21 of T. wilfordii and obtained 11 diterpene synthases with different functions. We further revealed that these diterpene synthases underwent duplication and rearrangement at approximately 2.3-23.7 million years ago (MYA) by whole-genome triplication (WGT), transposon mediation, and tandem duplication, followed by functional divergence. We first demonstrated that four key amino acids in the sequences of TwCPS3, TwCPS5, and TwCPS6 were altered during evolution, leading to their functional divergence and the formation of diterpene secondary metabolites. Then, we demonstrated that the functional divergence of three TwKSLs was driven by mutations in two key amino acids. Finally, we discovered the mechanisms of evolution and pseudogenization of miltiradiene synthases in T. wilfordii and elucidated that the new function in TwMS1/2 from the terpene synthase (TPS)-b subfamily was caused by progressive changes in multiple amino acids after the WGT event. Our results provide key evidence for the formation of diverse diterpenoids during the evolution of secondary metabolites in T. wilfordii.