Monitoring of sunitinib blood concentration and analysis of its safety and efficacy in Chinese patients with metastatic renal cell carcinoma / 中华泌尿外科杂志

Objective:To investigate the safety and efficacy of individualized sunitinib schedule for patients with metastatic renal cell carcinoma (mRCC) according to the monitoring results of plasma drug concentration.Methods:The clinical data of patients with mRCC who received sunitinib treatment in our center from January 2014 to December 2020 were retrospectively analyzed, including 20 patients who underwent monitoring of plasma drug concentration (monitoring group), and 45 patients, matched by propensity score matching, received sunitinib but did not undergo monitoring of plasma drug concentration during the same period (unmonitored group). In the monitoring group, there were 12 males and 8 females. The mean age was 52.9 years, and ECOG score ≤1 in 16 cases (80%). Three patients were in the IMDC favorable-risk group, 15 patients were in the intermediate-risk group, and 2 patients were in the high-risk group. There were 18 cases of clear cell carcinoma and 2 cases of non-clear cell carcinoma, 5 cases of ISUP grade 1-2 and 11 cases of grade 3-4. In the unmonitored group, there were 31 males and 14 females. The mean age was 57.7 years, and 30 patients had ECOG score ≤1, 15 cases ≥2. There were 10 cases in IMDC favorable-risk group, 23 cases in intermediate-risk group, and 12 cases in high-risk group. Thirty-seven cases were clear cell carcinoma and 8 cases were non-clear cell carcinoma, 8 cases were in ISUP grade 1-2 and 28 cases in grade 3-4. There were no statistically significant differences between the two groups in the above parameters ( P>0.05). The monitoring group used the regimen of taking sunitinib for 4 weeks and stopping for 2 weeks (4/2 week) in the first cycle. The blood concentration of sunitinib was monitored before the first cycle and on days 4, 7, 10, 14, 21 and 28, and personalized medication plan was formulated according to the curve of the blood concentration. The 4/2 week scheme was adopted in the undetected monitoring group.The two groups were compared in the incidence of adverse events (AEs), progression-free survival (PFS), overall survival (OS), tumor treatment response and other clinical outcomes. Results:In the monitoring group, 90% (18/20) of patients receiving sunitinib had a steady-state plasma concentration of more than 150ng/ml, of which 10 patients (50%) had a plasma concentration of 150-200 ng/ml and 8 patients (40%) had a plasma concentration of more than 200 ng/ml. Meanwhile, all patients with plasma concentration higher than 150 ng/ml developed severe AEs (grade 3 and above) after treatment. The other two patients' plasma concentration were 100-150 ng/ml, and did not have severe AEs.All patients in the monitoring group received individualized medication schedule adjustment according to the plasma drug concentration and the occurrence point of severe AEs, ensuring that the peak plasma drug concentration was maintained at about 100-150 ng/ml. Among them, 6 patients were changed to take 2 weeks and stop for 1 week (2/1 week schedule), 4 patients were changed to take 10 days and stop for 5 days (10/5 d schedule), 7 patients were changed to take 7 days and stop for 3 days (7/3 d schedule), and 3 patients were changed to take 5 days and stop for 2 days (5/2 d schedule). The incidence of severe AEs significantly decreased from 90% (18/20) to 35% (7/20), and the difference was statistically significant ( P=0.003), while the incidence of grade 3 and higher AEs was 55.6% (25/45) in the standard group, which was statistically significant compared with the incidence of severe AEs before adjustment in the monitoring group ( P=0.006). Further analysis of the efficacy difference between the two groups showed that the overall objective response rate in the monitoring group (40%, 8/20) was higher than that in the standard group (20%, 9/45), although the difference was not statistically significant ( P=0.09). Median PFS and OS were significantly longer in the monitored group than in the standard group (PFS: 23 vs. 10 months, P=0.002; OS: not reached vs.25 months, P=0.005). Conclusions:The bioavailability of sunitinib is high in mRCC patients, which may lead to higher plasma drug concentration, adjustment of medication regimen based on blood concentration monitoring significantly improved patient safety and clinical outcomes. However, further validation by larger-scale, multi-center and prospective studies is needed.

Development of the iGEM high school track / 生物工程学报

The international genetically engineered machine (iGEM) competition is a global top college academic competition in synthetic biology. The iGEM competition has exhibited extensive international influence and attracted teams from more than 40 countries and regions around the world to participate in. The annual iGEM outputs have attracted the attention of top academic journals or international media such as Science, Nature, Scientific American, The Economist, British Broadcasting Corporation (BBC), etc. High school teams participated in iGEM since 2011, and the number of high school teams has increased year by year. High school participants are increasingly becoming one of the most important forces to promote the development of iGEM and synthetic biology. IGEM competition has also become an important platform to foster the core literacy of high school students. This paper summarized the track rules, topic selection tendency and awards of high school teams based on data of 2017 to 2021 iGEM competition. In addition, we analyzed the significance of iGEM competition on fostering of high school students' core literacy and discussed the development trend of global high school teams, with the aim to provide a reference for high school team building in the future.

Effect of Jinshui Xiangsheng Prescription on Clinical Outcome of Advanced Prostate Cancer Based on Propensity Score Matching / 中国实验方剂学杂志

ObjectiveTo explore the effect of Jinshui Xiangsheng prescription on the five-year clinical survival outcome of patients with advanced prostate cancer. MethodFrom May 1, 2014 to May 1, 2016, patients with advanced prostate cancer from Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine and the Urology Department of the Second Affiliated Hospital of Nanjing University of Chinese Medicine were collected and treated with Jinshui Xiangsheng prescription (155 cases in the observation group). According to age and Gleason score, the patients without Jinshui Xiangsheng prescription were matched in a ratio of 1∶1 (155 cases in the control group). The androgen resistance rate, survival rate, median survival time, and median progress free survival time in 1, 3, 5 years were observed. The prognostic factors of advanced prostate cancer were analyzed and screened out by Chi-square test, t test, Kaplan-Meier and Cox survival analysis. ResultThe androgen resistance rates in the observation group in 1， 3， 5 years were 9.0% （14/155）， 72.3% （112/155）， and 92.9% （144/155）， respectively， and those in the control group were 20.6% （32/155）， 87.7% （136/155）， and 97.4% （151/155）. The 1-year （χ2=8.271，P<0.01）and 3-year （χ2=11.613，P<0.01） androgen resistance rates in the observation group were significantly lower than those in the control group. The median survival time and median progress free survival time in the observation group were （26.35±9.01） months and （11.02±4.40） months， respectively， and in the control group were （22.31±9.21） months and （9.87±5.12） months， respectively. The median survival time and median progress free survival time in the observation group were significantly longer than those in the control group （P<0.05， P<0.01）. The cumulative survival rates in 1， 3， 5 years in the observation group were 96.1% （149/155）， 80.6% （125/155）， and 39.4% （61/155）， respectively， and those in the control group were 94.2% （146/155）， 60.0% （93/155）， and 22.6% （35/155）， respectively. The 3-year （χ2=15.828，P<0.01） and 5-year （χ2=10.201，P<0.01） cumulative survival rates in the observation group were significantly higher than those in the control group. The monofactor analysis showed that the prognostic factors involved in Gleason score， initial prostate specific antigen （PSA）， tumor location， tumor stage， castration regimen， radiotherapy， chemotherapy， complete androgen blockade （CAB）， and Jinshui Xiangsheng prescription （P<0.05， P<0.01）. The multivariate analysis showed that initial PSA， tumor location， and tumor stage were the risk factors affecting the survival time of patients with advanced prostate cancer， whereas Jinshui Xiangsheng prescription， castration regimen， chemotherapy， radiotherapy， and CAB were the independent protective factors affecting the prognosis of advanced prostate cancer. ConclusionJinshui Xiangsheng prescription has a protective effect on the survival of patients with advanced prostate cancer， which reduces the androgen resistance rate and death risk of advanced prostate cancer， thus benefiting the survival of patients. Therefore， it deserves further promotion.

The impact of different metastatic sites on the prognosis of mRCC patients and its value for modification of International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) model / 中华泌尿外科杂志

Objective:To evaluate the impact of metastatic site on the prognosis in patients with metastatic renal cell carcinoma (mRCC), and it′s value for modifying the International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) criteria.Methods:The data of 218 patients pathologically diagnosed with mRCC were analyzed retrospectively in West China Hospital from Jan. 2009 to Dec. 2019. Among all patients, 71.6%(156/218) were male, and 89.0% (194/218) underwent nephrectomy. Most of the patients were pathologically diagnosed with renal clear cell carcinoma (176 patients, 80.7%). Lung (137/218, 62.8%) was the most observed metastatic site, following by bone (47/218, 26.1%), lymph node (37/218, 17.0%) and liver (23/218, 10.6%). All patients were classified into favorable (26 patients, 11.9%), intermediate (126 patients, 57.8%) or poor (37 patients, 17.0%) risk group according to IMDC criteria. Endpoints of this study were progression-free survival (PFS), overall survival (OS) and tumor response. The impact of metastatic sites on patients’ prognosis was analyzed, and those that had significant relationship with prognosis were then added into IMDC criteria and a modifying IMDC model was established. Predictive value of this model was further evaluated by calculating concordance index (C-index).Results:In the whole cohort, median PFS and OS were 13.0 and 33.0 months. Survival analysis suggested that patients with bone ( P=0.004), brain ( P=0.042) and liver ( P=0.046) had significantly shorter OS. Thus, patients were divided into two groups: patients with bone/brain/liver metastasis (82 patients, 37.6%) and patients with other metastatic sites (136 patients, 62.4%). Compared with patients with other metastatic sites, those who with bone/brain/liver metastasis had inferior tumor response by TKIs treatment (disease control rate: 51.2% vs. 73.5%, P=0.004). Multivariate analysis suggested that bone/brain/liver metastasis had negative impact on OS (25.0 vs. 47.0 mo, P=0.039). Furthermore, bone/brain/liver metastasis also showed significant relationship with shorter OS in IMDC low (30.0 vs. 62.0 months, P=0.036), intermediate (31.0 vs. 48.0 months, P=0.048) or high (7.0 vs. 18.0 months, P=0.037) risk group, indicating that metastatic site had predictive value for prognosis of mRCC patients. Based on that, bone/brain/liver metastasis were added into the IMDC criteria, and weighting each parameter was weighted according to its coefficient to patients’ OS. Finally, a modified IMDC scoring system were established. C-index of this modified model was 0.669 (0.599 for current IMDC criteria). Conclusions:Bone/brain/liver metastasis in mRCC patients indicated a shorter OS duration. When adding bone/brain/liver metastasis as a predictive parameter for prognosis of mRCC patients into IMDC criteria, the modified IMDC criteria could offer more accurate prediction for patients’ survival.

Vector Construction,Protein Expression,Purification,and Identification of Calmodulin Mutants N2 and C2 / 中国医科大学学报

Objective To construct expression vectors of calmodulin(CaM)mutants N2 and C2,and to express,purify,and identify the mutant proteins,in order to study the interactions between CaM and calcium channels. Methods The cDNA of N?lobe and C?lobe of CaM were used to prepare the cDNA of N2 and C2. Next,the recombinant cDNAs were cloned into a pGEX?6p?3 plasmid,and the recombinant plasmids were trans?ferred into E.coli BL21 cells. The transfected BL21 cells were stimulated with IPTG. The fusion proteins were extracted by ultrasonication and puri?fied by using GS?4B beads. Finally,protein activity was identified by the pull?down assay. Results Both the restriction digestion map and the DNA sequence identification results confirmed that the recombinant plasmids were successfully constructed. SDS?PAGE results showed high purity and concentration of N2 and C2 proteins. Their activities and binding abilities with the calcium channel fragment were confirmed by the pull?down assay.Conclusion In this study,expression vectors of N2 and C2 are successfully constructed,and physiologically active N2 and C2 CaM mutant proteins are obtained.

Current aging research in China

The mini-review stemmed from a recent meeting on national aging research strategies in China discusses the components and challenges of aging research in China. Highlighted are the major efforts of a number of research teams, funding situations and outstanding examples of recent major research achievements. Finally, authors discuss potential targets and strategies of aging research in China.

Effect of aging on repair capability of lung stem cells / 中国比较医学杂志

Objective To study the impact of aging on the capability of lung stem cell steady-state maintaining and bronchial epithelial cells regeneration and differentiation during the repair of lung epithelial cells after naphthalene induced bronchial epithelialium injury.Methods The proportion of lung stem cells in mice after naphthalene treatment was analyzed by immunohistochemistry and FACS.The repair efficiency of lung epithelial cell in young and old mice was examined by immunohistochemistry staining and FACS.Results The data suggested that aging didn ’ t change the proportion of lung stem cells ( including the distal lung epithelial stem cells/progenitor cells and lung mesenchymal stem cells/progenitor cells) under normal physiological conditions.After naphthalene injury, more serious injury and decreased repairing capacity was observed in old group.Lung progenitor cells /total lung cells decreased during the repair process of lung bronchial epithelialium ( clara cell) injury.The ratio of regenerated cell to lung progenitor and stem cells were also significantly decreased in old group.Conclusion The regenerated capability of lung stem cells after lung bronchial epithelialium injury decreased with aging.This might be the reason of more incidence of lung injury and worse therapeutic results in the elder in clinic.

Effect of different isolations for nucleic acid and protein complex on affinity of enriched library in SELEX experiment / 第三军医大学学报

Objective To compare the effect of different partition of nucleic acid protein complex on the affinity of enriched library in systematic evolution of ligands by exponential enrichment (SELEX). Methods Two protocols were adopted to select the enriched library to transforming growth factor ? receptor Ⅱ(TGF ? RⅡ). Protocol 1: protein was absorbed on the surface of 96 well plate; then, selection was carried out; the binding nucleotide acids were eluted from the supporter directly. Protocol 2: selection was done in solution; nucleotide acid protein complex was captured in nitrocellulose membrane; the binding nucleotide acids were obtained from membrane. Filter biding assay and gel shift assay were performed to detect the affinity of the enriched ssDNA library from different protocols. Results After 4 rounds of selection, the affinity to TGF ? RⅡ was obviously improved in the enriched library from protocol 2 compared with the initial library, while no such improvement was found in the enriched library from protocol 1. Conclusion In the SELEX experiment, the way of selection in solution, then partition of the binding nucleotide acids in filter is easier to enrich the binding fragment from initial ssDNA random library, compared with the way of fixation of target protein to solid supporter, then selection between the solid phase and liquid phase and elution of the binding nucleotide acids from supporter.

Screening of nucleic acids affinity to recombinant human TGF-? RⅡ / 第三军医大学学报

Objective To select the aptamer to an extracellular soluble fragment of recombinant human TGF-? receptor Ⅱ (TGF-? RⅡ) in order to antagonize TGF-? effectively by using systematic evolution of ligants by exponential enrichment (SELEX). Methods Initial random RNA library was transcripted in vitro from ssDNA 5′-TAATACGACTCACTATAGGGAGGACGATGCGG-N60-CAGACGACTCCCCGA-3′; rhTGF-? RⅡ was used as target protein. Totally,selection of 8 times was carried out in SELEX experiment. Membrane binding assay was performed to detect the evolution of enriched RNA library; Electrophoretic mobility shift assay (EMSA) was done to determine the affinity between the selected nucleic acid sequence and TGF-? RⅡ. Results Evolution of the enriched RNA library along the increased affinity to TGF-? RⅡ was observed with the development of selection. Two types of dominant sequences were isolated and named as sequence A and sequence B. In membrane binding assay,both sequences A and B showed obvious affinity to TGF-? RⅡ. However,no retarded bands were seen in EMSA. Conclusion The affinity of sequences A and B to TGF-? RⅡ is beyond satisfaction. However,possible sequences with improved affinity to TGF-? RⅡ can be selected by post-SELEX on the basis of candidate sequences A and B.

Expression of positive and negative regulators of cell cycle during wound healing / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To detect the expression of cell cycle positive regulators cyclin D(1), cyclin E, CDK(2), CDK(4) and negative regulators p21(cip1), p27(kip1), p16(ink4a) and p15(ink4b) during wound healing in rats.</p><p><b>METHODS</b>Open wounds of full-thickness skin, diameter 1.8 cm, on rat backs were used as the wound model. Wound tissues were harvested on postwounding days 3, 5, 7, 9, 11, 14, 21 and 30. Ki67 expression in granulation tissue was detected by immunohistochemical assay. The patterns of the expression of cyclin D(1), cyclin E, CDK(2), CDK(4) and p21(cip1), p27(kip1), p16(ink4a), p15(ink4b) were detected by Western blot.</p><p><b>RESULTS</b>Cell proliferation in granulation tissue took place predominantly within the first week after injury, with the proliferation peak occurring at postwounding day 5. There were no dramatic variations in the expression of cyclin D(1), CDK(2) and CDK(4) during wound healing. Up-regulated cyclin E was maintained from day 3 to 11 after injury, and then was down-regulated. No expression of p16(ink4a) and p15(ink4b) was found. p21(cip1) was expressed only from day 7 to 14, with peak expression observed on day 9. Constitutive p27(kip1) was expressed throughout wound healing with low levels in the proliferating period of day 3 to 5 and with increased levels in the post-mitotic and remodeling stage. The expression of p21(cip1) and p27(kip1) showed an inverse gradient to that of Ki67.</p><p><b>CONCLUSION</b>p21(cip1) and p27(kip1) play a supervising role in preventing the hyperproliferative tendency in tissue repair.</p>

A CTX-M extended-spectrum ?-lactamases-producing Klebsiella pneumonia isolated in Beijing / 中华检验医学杂志

Objective To determine the ESBLs of Klebsiella pneumoniae and ESBL typing by molecular genetic procedures. Methods MIC test and ESBLs confirmation test were taken by agar dilution method in 80 strains of Klebsiella pneumoniae isolated in Beijing area from July 1999 to December 1999. To the 20 ESBLs positive conjugates isoelectric focusing was given. And nucleotide sequencing was analysed to conjugate strain CK23. Results 28 (35%) of the 80 strains of Klebsiella pneumoniae produced ESBLs. In the 28 strains, 20 were successfully done with conjugation of resistant plasmid. Isoelectric focusing results revealed that 13 (65%) of the 20 strains produced an ESBLs protein with a pI of 8.8 with or without additional pI 7.6 and pI 5.4. These ? lactamases were all inhibited by clavulanite acid. The strains with pI=8.8 protein were highly resistant to cefotaxime and ceftriaxone but were susceptible or intermediate to ceftazidime. We picked CK23 strain out from 13 conjugates for gene cloning by the primers designed for bla CTX M and nucleotide sequencing. The results showed that the ESBLs gene in CK23 was CTX M, highly similar to CTX M 3 but had 3 amino acids, which were Glu39Gly, Leu122Pro, Asp278His. Conclusions 28 (35%) of the 80 strains of Klebsiella pneumoniae produced ESBLs. Resistant plasmid successfully conjugated in 20 of the 28 ESBLs producing strains. 13 of the 20 (65%) conjugates pruduced a pI =8.8 ESBLs,which was CTX M ESBLs,with only 3 amino acids different from CTX M 3. This paper indicated that a type of CTX M ESBLs existed in Klebsiella pneumoniae in Beijing, which specifically confers resistance to cefotaxime.

Expression of three S100A calcium-binding proteins in human gastric cancer and non-cancerous mucosa as detected by QRT-PCR / 解放军医学杂志

Objective To verify the authors' previous cDNA micro-array results and to further investigate the moleculer mechanisms of gastric cancer. Methods Quantitative real-time RT-PCR was employed to detect the expressions of three S100A calcium-binding genes in 22 fresh surgical samples of gastric tumor tissue and non-cancerous mucosa from the same patients. Results The transcription level of S100A2 in primary cancer lesion was elevated in 80% of samples when compared with matching non-neoplastic mucosa (P=0.018) and the average up-regulation level was 10.78 fold. 55% of cancer lesions showed higher transcription level of S100A4 than their adjacent non-neoplastic mucosa, the average up-regulation level was 2.31 fold. S100A6 transcription level was higher in 74% (P=0.01) of primary cancer lesion with an 2.25 fold up-regulation than the adjacent non-neoplastic mucosa. After rectified by ?_2-microglobulin, the relative expression levels of S100A2, S100A4 and S100A6 were 2.83?10~ -4 , 6.44?10~ -2 and 0.41, respectively. According to the Spearman correlation coefficient analysis there were significant positive correlations between S100A2 and S100A4, and S100A2 and S1006 (P value were 0.00 and 0.017, respectively). Conclusion The changes in S100A2 and S100A6 genes may be an early event in a majority of gastric cancer patients, while S100A4 may be associated with the infiltration of gastric cancer. Further study on the three genes might be helpful for understanding the nature of gastric carcinoma.

Changes of some seminal biochemical indices in chronic prostatitis / 第三军医大学学报

The activities of acid phosphatase and ?-1,4 glucosidase and the concentration of citric acid in the semen of 51 cases of chronic prostatitis were determined.The mean value was 694.33?346.61 Bodansky units for acid phosphatase,43.84?24.70mIU/ml for ?-1,4 glucosidase,and 3.26 ?2.37 mg/ml for citric acid.The activity of acid phospohatase and the concentration of citric acid were significantly lower in the patients with chronic prostatitis than in the normal subjicts while the activity of ?-1,4 glucosidase showed little changes.The findings suggest that the functions of the prostate was severely imparired and that of the epididymis remained normal in patients with chronic prostatitis.

Studies on correlation between S100A4 expression and biological behavior and prognosis of gastric cancer / 解放军医学杂志

Objective To study the expression of S100A4 in gastric cancer and normal gastric tissue, and analyze its correlation with the clinico-pathological features and prognosis of gastric cancer. Methods Real-time quantitative reverse transcription PCR (real time qRT-PCR) was used to detect the S100A4 expression in 20 fresh surgical samples of gastric cancer and normal gastric tissue as controls. The microarray of gastric cancer tissue was established for the analysis of the S100A4 expression immunohistochemically in 208 gastric cancer tissue and isogeneic normal gastric mucosa and lymph node with metastasis. Results The S100A4 expression was increased in 55% (11/20) of gastric cancer samples with an average of 2.31 fold up-regulation of that of the normal mucosa. Patients with lymph node metastasis showed a higher percentage of elevated S100A4 transcription than those without metastasis (P=0.024). As displayed by immunohistochemistry, the positive rate of S100A4 in non-neoplastic mucosa, primary tumor and lymph node with metastasis was 9.4%, 28.1% and 32.2%, respectively (P≤0.01). A higher percentage of elevated S100A4 expression was shown in patients in advanced stage than in patients in early stage (P=0.004). In primary tumor lesions, the S100A4 expression correlated significantly with the depth of invasion (P=0.003) and poorer prognosis (P=0.034). S100A4 expression in lymph node with metastasis was also associated with poor outcome (P=0.002). Multifactorial Cox's regression analysis showed that TNM stage (P=0.029) and the expression levels of S100A4 (P=0.024) in lymph node were independent influence factors for prognosis. Conclusions Expression of S100A4 may be a late event which is associated with the progression and prognosis of gastric cancer. The analysis of S100A4 expression in lymph-node metastasis is helpful in judging the prognosis of gastric cancer.