鼠胚成纤维细胞的培养条件及滋养层制备

背景:鼠胚成纤维细胞经丝裂霉素C处理或Y射线照射阻断其有丝分裂后,铺层的细胞可保持活力但不增殖,并能够在生长过程中产生促进胚胎十细胞生长的因子和抑制胚胎干细胞分化的因子,但其生命期有限.目的:探讨分离小鼠胚胎成纤维细胞的最适胚龄与培养条件,以及用其制备细胞饲养层的效果.设计、时间及地点:细胞学体外观察,于2005-11/2006-07在广东省计划生育专科医院实验室完成.材料:清洁级昆明系孕7.5,10.5,13.5,16.5,19.5 d雌鼠各5只.方法:无菌取上述各孕龄小鼠胚胎分离鼠胚成纤维细胞,调整细胞浓度为1×107L-1、1×109L-1 1×1011 L-1,胰酶消化传代.当胚胎成纤维细胞生长并相互接触时,加入丝裂霉素C作用2-4 h,用无钙无镁PBS配制的胰蛋白酶液消化2～5 min,终止后用吸管吹打皿底,使细胞充分分离,以含10%胎牛血清的DMEM液调整细胞浓度为5×108 L-1,将该悬液移入明胶处理的培养皿内常规培养,制备鼠胚成纤维细胞饲养层.主要观察指标:胎龄、细胞浓度、传代次数对鼠胚成纤维细胞生长增殖的影响.不同胎龄鼠胚成纤维细胞饲养层制备效果.结果:7.5～19.5 d鼠胚均可分离出成纤维细胞,但7.5 d.10.5 d鼠胚分离所得成纤维细胞数最少,寿命短,且混有大量杂细胞:13.5d鼠胚分离所得成纤维细胞数量多,增殖快,所含杂细胞少:16.5～19.5 d鼠胚分离所得成纤维细胞生长状态不良,增殖缓慢,杂细胞较多.同等条件下与1x107 L-1、1×1011 L-1浓度比较,1×109 L-1浓度的鼠胚成纤维细胞生长状况良好,铺层时间适中,细胞寿命最长(P＜0.05).以13.5 d鼠胚成纤维细胞经初代培养后传4代,成功建立了成纤维细胞株,相同条件下1～代细胞形态,体积、生长状况无明显差别.7.5～19.5 d鼠胚成纤维细胞制备的饲养层,铺层时间基本相似(P＜0.05),细胞寿命均可维持一二周.结论:分离小鼠胚胎原代成纤维细胞的最适胎龄及细胞浓度分别为13.5d与1×109 L-1,4代以内传代次数不影响鼠胚成纤维细胞的生长增殖,且不同胎龄所制备的饲养层细胞寿命无差别.     

补肾健脾化瘀法治疗男性骨质疏松症疼痛的效果

目的:观察补肾健脾基础上加强化瘀药物制成补肾壮骨中药对男性骨质疏松患者疼痛程度的影响。方法:①选择1998-07/2004-07解放军广州军区广州总医院住院及门诊就诊的男性老年性骨质疏松症患者198例,年龄65～83岁。符合骨质疏松症综合诊断评分标准,且均对治疗方案知情同意。②按随机数字表法将患者分为2组;补肾壮骨中药组101例,降钙素组97例。补肾壮骨中药组:均囗服补肾壮骨中药(由生地、山药、山萸肉、泽泻等药物组成,广东一方制药厂生产,批号:980705,100g/包),1包/次,2次/d。降钙素组:均肌肉注射降钙素(诺华公司产品,生产批号980823,990523,011121,50IU/支),其中第1周100IU,1次/d,第2,3,4周100IU,每周2次。两组均治疗4周。③分别于治疗前,治疗1,2,3,4周后及停药后2周记录疼痛强度(共计0～10,0为无痛,10为剧烈疼痛)、疼痛缓解度(共计0～4度,0度为无缓解,4度为完全缓解),计算镇痛有效率:显效(明显缓解 完全缓解),有效(中度缓解),总有效率(显效 有效)。④计量和计数结果差异比较分别采用t检验和χ2检验。结果:男性老年性骨质疏松症患者198例均进入结果分析。①骨质疏松症疼痛强度:补肾壮骨中药组治疗后2,3,4周和停药后2周和降钙素组治疗后1,2,3,4周和停药后2周疼痛强度明显低于治疗前(t=2.35～2.93,P<0.05),补肾壮骨中药组治疗1和2周后疼痛强度明显高于降钙素组(5.32±1.21,3.26±1.32;3.49±1.39,2.21±1.28,t=2.98,2.36,P<0.05)。②骨质疏松症疼痛缓解情况:治疗2周后,补肾壮骨中药组完全缓解率明显低于降钙素组(5.9%,18.6%,χ2=6.25,P<0.05)。补肾壮骨中药组及降钙素组治疗1,3,4周后和停药后2周疼痛完全缓解率、明显缓解率、中度缓解率差异不明显(P>0.05)。③镇痛有效率:治疗2周后,补肾壮骨组镇痛显效率明显低于降钙素组(44.6%,58.8%,χ2=5.721,P<0.05),有效率和总有效率与降钙素组相近(P>0.05)。补肾壮骨中药组和降钙素组治疗4周后和停药后2周镇痛总有效率、显效率、有效率比较,差异不明显(P>0.05)。结论:补肾壮骨中药可明显缓解男性患者骨质疏松症疼痛,作用效果基本与降钙素相当。     

Diagnostic Accuracy of the Apparent Diffusion Coefficient for Microvascular Invasion in Hepatocellular Carcinoma: A Meta-analysis

Background and AimsMicrovascular invasion (MVI) is a major risk factor for the early recurrence of hepatocellular carcinoma (HCC) and it seriously worsens the prognosis. Accurate preoperative evaluation of the presence of MVI could greatly benefit the treatment management and prognosis prediction of HCC patients. The study aim was to evaluate the diagnostic performance of the apparent diffusion coefficient (ADC), a quantitative parameter for the preoperative diagnosis MVI in HCC patients.MethodsOriginal articles about diffusion-weighted imaging (DWI) and/or intravoxel incoherent motion (IVIM) conducted on a 3.0 or 1.5 Tesla magnetic resonance imaging (MRI) system indexed through January 17, 2021were collected from MEDLINE/PubMed, Web of Science, EMBASE, and the Cochrane Library. Methodological quality was evaluated using Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2). The pooled sensitivity, specificity, and summary area under the receiver operating characteristic curve (AUROC) were calculated, and meta-regression analysis was performed using a bivariate random effects model through a meta-analysis.ResultsNine original articles with a total of 988 HCCs were included. Most studies had low bias risk and minimal applicability concerns. The pooled sensitivity, specificity and AUROC of the ADC value were 73%, 70%, and 0.78, respectively. The time interval between the index test and the reference standard was identified as a possible source of heterogeneity by subgroup meta-regression analysis.ConclusionsMeta-analysis showed that the ADC value had moderate accuracy for predicting MVI in HCC. The time interval accounted for the heterogeneity.     

Pharmacokinetics,Bioequivalence and Safety of Cloperastine in Chinese Healthy Subjects Under Fasting and Postprandial Conditions

BackgroundCloperastine is a pivotal antibechic widely prescribed to treat cough caused by respiratory diseases. The present trial evaluated the pharmacokinetics (PK), bioequivalence (BE) and safety effects of the generic test (T) tablet of cloperastine after single-dose administration of cloperastine, compared with the original reference (R) tablet of cloperastine.ObjectiveThe purpose of this trial was to compare the PK, BE and safety of a test 10 mg versus the reference 10 mg formulation of cloperastine under fasting and postprandial conditions in healthy Chinese volunteers.MethodsA single-centre, randomised, open, double-cycle, self-crossover, single oral administration Phase I trial was performed in healthy Chinese volunteers. A total of 60 subjects were enrolled in either the fasting (28 subjects) or the postprandial condition (32 subjects). Subjects randomly received a single dose of the T or R preparation (10 mg dose). Plasma concentrations of cloperastine were analysed by a validated LC-MS/MS method. The primary endpoints of the PK parameters were the area under the plasma concentration-time curve from zero to 72 h (AUC_0–72h), under the plasma concentration-time curve from zero to infinity (AUC_0–∞) and the maximal plasma concentration (C_max). The equivalence standard range (80.0–125.0%) was used to evaluate the BE of the two preparations. The safety parameter as secondary endpoint was mainly evaluated by the occurrence of adverse events (AEs).ResultsA total of 25 and 30 subjects in the fasting and postprandial conditions completed this clinical trial, respectively. The geometric mean ratio (GMR) of the T/R for the C_max, AUC_0–72h and AUC_0–∞ were 102.1%, 103.8% and 104.0% in the fasting condition, respectively. In the postprandial condition, the GMR of the T/R for the C_max, AUC_0–72h and AUC_0–∞ were 94.2%, 98.8% and 99.0%, respectively. All the values fell within the range (80.0–125.0%). The C_max and AUC_0–72h values of the T and R preparations in fasting and postprandial conditions were not statistically significant (P > 0.05). Furthermore, no serious adverse events (SAEs) occurred during the whole trial.ConclusionsThe T and R preparations were bioequivalent under both conditions. Food has no significant effect on the absorption of cloperastine. Moreover, T and R preparations were well tolerated. The trial registration number (TRN) and date of registrations were CTR20212515, 13 October 2021.Supplementary InformationThe online version contains supplementary material available at 10.1007/s40268-022-00406-2.     

A Novel Peptide from Polypedates megacephalus Promotes Wound Healing in Mice

Amphibian skin contains wound-healing peptides, antimicrobial peptides, and insulin-releasing peptides, which give their skin a strong regeneration ability to adapt to a complex and harsh living environment. In the current research, a novel wound-healing promoting peptide, PM-7, was identified from the skin secretions of Polypedates megacephalus, which has an amino acid sequence of FLNWRRILFLKVVR and shares no structural similarity with any peptides described before. It displays the activity of promoting wound healing in mice. Moreover, PM-7 exhibits the function of enhancing proliferation and migration in HUVEC and HSF cells by affecting the MAPK signaling pathway. Considering its favorable traits as a novel peptide that significantly promotes wound healing, PM-7 can be a potential candidate in the development of novel wound-repairing drugs.     

An Innovation of the Markov Probability Model for Predicting the Remaining Service Life of Civil Airport Rigid Pavements

In view of the time series update of airport runway health status detection data, the Markov chain of stochastic process theory was adopted. Considering the influence of aircraft traffic load, age, and pavement structure surface-layer thickness on the performance deterioration process of airport runways, the method of survival analysis was used. The parameter model of survival analysis was used to establish the duration function model of the four condition states of the airport runway PCI (pavement condition index). The Markov transition matrix for the performance prediction of airport runways was constructed. In order to evaluate the ability of the Markov transition matrix method to predict the trend of deterioration for PCI of the airport runway under different conditions of aircraft traffic volume and thickness of the runway pavement surface, a data set was constructed with the actual inspection data of the airport runway, and the corresponding samples were selected for analysis. The results showed that a Markov transition matrix for airport runway performance prediction, constructed based on survival analysis theory, can combine discontinuous inspection data or monitoring data with Weibull function survival curves. The method proposed in this paper can quantitatively predict the remaining service life of airport runways and provide support for cost-effective decisions about airport pavement maintenance and rehabilitation.