Enzyme enhancement for the measurement of protein C

Protein C measurement is now a necessary work-up of a patient with thrombosis. We described an enzyme enhancement of Laurell's immunoelectrophoresis for assay of protein C antigen. With this modification, the rockets are well defined and easily visualized and the sensitivity of the assay increased (2.5%). Samples with low protein C antigen are easily assayed.     

Anatomical Variations in Stylopharyngeus Muscle Insertions Suggest Interindividual and Left/Right Differences in Pharyngeal Clearance Function of Elderly Patients: A Cadaveric Study

The stylopharyngeus plays a critical role in the clearance of the piriform recess. We dissected 78 sides of the pharynx from 55 donated cadavers and observed histology of another seven sides of the pharynx from seven cadavers. The stylopharyngeus consistently comprised (1) a descending muscle bundle surrounding the piriform recess and (2) an additional short sheet inserting into the tonsillar bed. Histologically, the former bundle connected to a thick fascia providing the lateral glossoepiglottic fold, extending along the submucosa of the piriform recess, and covering the thyroid cartilage, whereas the latter sheet intermingled with other pharyngeal wall muscles at and near the tonsillar bed. Notably, in 44.4% of female specimens, the additional sheet occupied a greater proportion in cross section than the descending muscle bundle. Given the different directions, the additional sheet seemed to check clearance function of the descending bundle for the piriform recess. Thus, particularly in women, interindividual differences in pharyngeal clearance were likely to depend on whether the additional sheet is strong or weak. Chin down in combination with tilting and rotating the head may represent effective exercises of the stylopharyngeus that could compensate for the disadvantages of additional insertion.     

Standardized <Emphasis Type="Italic">Citrus unshiu</Emphasis> peel extract ameliorates dexamethasone-induced neurotoxicity and depressive-like behaviors in mice

Dried Citrus unshiu peel, also known as Chinpi, have been commonly used as a traditional medicine to improve for allergy, inflammation and hepatopathy. Many previously studies have reported that citrus flavonoids show neuroprotective activities. However, the antidepressant-related effects of C. unshiu peels have not been well characterized. Here, the antidepressant-like effects of standardized C. unshiu peel extract (SCP) were evaluated in in vivo and in vitro depression models induced by dexamethasone (DEX), a synthetic glucocorticoid. Male ICR mice (9-week-old) were injected the DEX (40 mg/kg) and were orally given SCP daily (30, 100, and 300 mg/kg) for 14 consecutive days. The depressive-like behaviors were determined by use of open filed test (OFT), sucrose preference test (SPT), tail suspension test (TST), and forced swim test (FST). We show that treatment with SCP significantly alleviated DEX-induced depressive-like behaviors and reduced neurotoxicity in a concentration dependent manner in SH-SY5Y cells. Additionally, repeated DEX injection markedly decreased brain derived neurotrophic factor (BDNF) level, tropomyosin receptor kinase B (TrkB), and cyclic AMP-response element-binding protein (CREB), while SCP treatment improved these levels in the cerebral cortex and hippocampus regions. Our findings suggest that SCP exhibits significant antidepressant-like effects in the DEX-induced depressive animal model, and this activity may be mediated by preventing corticosterone-induced neurotoxicity.     

Genetic insight into cytochrome P450 in Chinese from the Chinese Millionome Database

Genetic variations of cytochrome P450 (CYP) influence the inter‐individual differences in drug response. Here, we collected 8682 variants of 57 CYP genes and cytochrome P450 oxidoreductase (POR) from a large‐scale sequencing project in Chinese, Chinese Millionome Database (CMDB). In addition, 52 294 variants from the Genome Aggregation Database (gnomAD) had been simultaneously identified and analysed. Rare variants with a variant allele frequency (VAF) < 0.01 comprised 41.4% (3594/8682) of identified variations in the CMDB, while 98.1% (51 320/52 294) in the gnomAD were rare. Out of 8682 variants in the CMDB, 66.9% (5808/8682) were in introns and only 4.3% (377/8682) were missense variants. In contrast, 36.2% (18 929/52 294) variants in the gnomAD were missense. The common alleles with a VAF over 0.1 were found in CYP1A2*1C, CYP1A2*1F, CYP2C19*2, CYP2D6*2, CYP2D6*10, CYP3A5*3 and CYP4F2*3, with a VAF of 0.161, 0.6, 0.27, 0.274, 0.678, 0.92 and 0.233, respectively. The growing number of genetic variations in CYP genes as more genomes are sequenced would increase the power to predict drug metabolism and response based on the genotype of the particular individual.     

Antitumour properties based on the self‐assembly of camptothecin and carbamoylmannose conjugates

Camptothecin (CPT) and its analogues show potent antitumour activity. However, poor water solubility and severe side effects have restricted their applications in clinical practice. In this paper, a novel self‐assembly based on camptothecin and carbamoylmannose conjugates ( CPT‐Man ) was constructed. The self‐assembly increased the water solubility of camptothecin to 0.64 mg/ml and antitumour activity. Moreover, CPT‐Man could induce obvious cancer cell apoptosis. This work provides a new approach for exploring carbohydrate‐modified antitumour properties by self‐assembled CPT drugs.     

贝伐珠单抗联合伊立替康与雷替曲塞治疗氟尿嘧啶耐药晚期结直肠癌的疗效和安全性

目的 观察贝伐珠单抗联合伊立替康与雷替曲塞方案在氟尿嘧啶类药物耐药后的晚期结直肠癌患者中的疗效及安全性。方法 收集中国医科大学附属第一医院2014—2019年收治的氟尿嘧啶类耐药的60例晚期结直肠癌患者,对照组30例,应用伊立替康联合雷替曲塞方案（IR）;实验组30例,应用IR联合贝伐珠单抗方案。分析比较两组患者的客观有效率（ORR）、疾病控制率（DCR）、无进展生存时间（PFS）及不良反应发生情况。结果 实验组和对照组ORR分别为6.67%和3.33%,DCR分别为66.67%和53.33%,两组ORR和DCR比较差异无统计学意义。实验组和对照组中位PFS分别为6.0个月和3.1个月,差异有统计学意义（P=0.020 4）。两组不良反应以Ⅰ/Ⅱ级多见,Ⅲ/Ⅳ级不良反应发生率低,实验组蛋白尿的发生率高于对照组,差异有统计学意义（P=0.001）。其余如出血、转氨酶升高、恶心、呕吐、腹泻、发热、皮疹、高血压等不良反应发生率均为实验组高于对照组,但差异无统计学意义。结论 贝伐珠单抗联合伊立替康与雷替曲塞方案可提高既往氟尿嘧啶类治疗耐药后的晚期结直肠癌患者的疗效,无进展生存期增加,不良反应可耐受,值得进一步研究。     

妊娠期哮喘患者疾病管理及用药情况的调查分析

目的了解妊娠期哮喘患者哮喘控制及药物使用的情况,分析妊娠期哮喘控制不佳的原因。方法通过对41例诊断为哮喘的妊娠期女性进行问卷调查,收集其目前哮喘控制水平、药物使用情况、吸入装置与吸入技巧、用药依从性、对疾病和药物的认识、影响哮喘控制的因素等情况,根据答题结果分析。结果根据哮喘控制问卷评分,"良好控制"占39.0%;"部分控制"+"未控制"占61.0%。在妊娠期间,34.1%患者有过至少1次因哮喘急性发作门/急诊就诊或入院治疗。20.0%患者能正确吸入药物,52.6%患者用药依从性较好。39%患者认为吸入激素对胎儿危害会大于哮喘,61.0%患者分不清楚缓解药和控制药。影响哮喘控制原因中,48%患者担忧激素会对胎儿影响,无患者选择经济因素。结论妊娠期哮喘控制欠佳,药师参与妊娠哮喘患者管理有望提高其控制水平。     

Thromboelastography (TEG) in normal pregnancy and its diagnostic efficacy in patients with gestational hypertension,gestational diabetes mellitus,or preeclampsia

BackgroundThromboelastography (TEG) provides global assessment of hemostatic function and has been recommended to monitor potential coagulopathies during pregnancy in which hypercoagulable state is favored. In present study, we established the reference intervals (RIs) of the TEG parameters (R, K, MA, and α‐angle) with Chinese pregnant women of third trimester. In addition, we examined the diagnostic efficacies of the TEG parameters in the patients diagnosed of gestational hypertension (GH), gestational diabetes mellitus (GDM), or preeclampsia (PE).MethodsWith specified including and excluding criteria, non‐pregnant controls, healthy pregnant women, and pregnant women with GH, GDM, or PE had their venous blood drawn at Beijing Obstetrics and Gynecology Hospital, followed by TEG tests performed in the clinical laboratory.ResultsThe RIs determined with the healthy pregnant women (in third trimester) for R, K, MA, and α‐angle were 4.0‐7.7, 1.2‐3.2, 51.9‐70.1, and 41.4‐74.4, respectively. When compared with the healthy pregnancy group, the K value was significantly decreased in GH patients but increased in PE patients; MA was significantly lower in the PE group. In the receiver operating characteristic curve (ROC) analyses, K value was able to efficiently distinguish normal pregnancy from the GH patients, with an AUC of 0.86 which is far better than those of R (AUC = 0.57) and MA (AUC = 0.56). For the PE patients, the AUC of MA (0.69) was significantly greater than that of R (0.50).ConclusionsThromboelastography may provide more accurate experimental basis for monitoring coagulation functions especially in pregnant women with complications of GH and PE.     

Development of novel quality control material based on CRISPR/Cas9 editing and xenografts for MLH1 protein deficiency testing

BackgroundMismatch repair deficiency (dMMR) status induced by MLH1 protein deficiency plays a pivotal role in therapeutic decision‐making for cancer patients. Appropriate quality control (QC) materials are necessary for monitoring the accuracy of MLH1 protein deficiency assays used in clinical laboratories.MethodsCRISPR/Cas9 technology was used to edit the MLH1 gene of GM12878Cas9 cells to establish MLH1 protein‐deficient cell lines. The positive cell lines were screened and validated by Sanger sequencing, Western blot (WB), and next‐generation sequencing (NGS) and were then used to prepare formalin‐fixed, paraffin‐embedded (FFPE) samples through xenografting. These FFPE samples were tested by hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) for suitability as novel QC materials for MLH1 protein deficiency testing.ResultsWe successfully cultured 358 monoclonal cells, with a survival rate of 37.3% (358/960) of the sorted monoclonal cells. Through Sanger sequencing, cell lines with MLH1 gene mutation were identified. Subsequently, two cell lines with MLH1 protein deficiency were identified by WB and named as GM12878Cas9_6 and GM12878Cas9_10. The NGS results further confirmed that the MLH1 gene mutation in these two cell lines would cause the formation of stop codons and terminate the expression of the MLH1 protein. The H&E staining and IHC results also verified the deficiency of the MLH1 protein, and FFPE samples from xenografts proved their similarity and consistency with clinical samples.ConclusionsWe successfully established MLH1 protein‐deficient cell lines. Followed by xenografting, we developed novel FFPE QC materials with homogenous, sustainable, and typical histological structures advantages that are suitable for the standardization of clinical IHC methods.