Comparative study of the effects of three semen preparation media on semen analysis,DNA damage and protamine deficiency,and the correlation between DNA integrity and sperm parameters

Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media,such as PureSperm （Nidacon,Gothenburg,Sweden）,Sil-Select Plus^TM （Fertipro,Beemem,Belgium） and SpermGrad^TM（Vitrolife,Gothenburg,Sweden）. The differences in mean percentages of semen parameters were assessed by repeated measures analysis. Correlations of sperm DNA damage,as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay,and of protamine deficiency,as measured by chromomycin A3 （CMA3） staining with sperm parameters,were determined by Pearson＇s correlation. After preparation with all three media,sperm concentrations decreased （P〈0.05） while percentages of sperm with normal morphology increased （P〈0.05）. Percentages of sperm motility,rapid motility and progressive motile concentration （PMC） increased （P〈0.05） for each ofthese parameters,PureSperm preparation gave the best results （P〈0.05）. The percentage of DNA damage decreased in the PureSperm and Sil-Select Plus preparations （17.9% and 31.3%,respectively,P〈0.05） and increased in the SpermGrad preparation （56.3%,P〈0.05）. Protamine deficiency also decreased in all three kinds of media,59.3%,47.7% and 40.3% for PureSperm,Sil-Select Plus and SpermGrad preparations,respectively （P〈0.05）. The percentage of DNA-damaged sperm was negatively correlated with the percentages of sperm motility,rapid motility and PMC,but was positively correlated with static motility （P〈0.05）. This comparative study and correlation analysis revealed that PureSperm preparation yielded sperm with the best motility and the lowest percentage of protamine deficiency. The Sil-Select Plus preparation yielded sperm with the lowest amount of DNA damage. The SpermGrad preparation had a high percentage of sperm with normal morphology,but also had the highest percentage of sperm with DNA damage. Sperm DNA damage was correlated with percentages of sperm motility,rapid motility,static motility and PMC.

Emergence of lesions outside of the basal ganglia and irreversible damage to the basal ganglia with severeβ-ketothiolase deficiency:A case report

BACKGROUNDβ-ketothiolase deficiency(β-KTD)is an inherited disease,and insufficient attention has been paid to imageology due to its lower morbidity.Therefore,few lesions outside the basal ganglia have been found before,and the persistent pathological changes have rarely been reported.CASE SUMMARY A 10-mo-old Chinese female patient with a free previous medical history but with poor physical and athletic development had received the haemophilus influenzae vaccine and then developed a low fever 2 d prior.She was initially diagnosed with severe brain injury,central respiratory failure,metabolic acidosis complicated with respiratory alkalosis,hyper-IgE,etc.With further examination,a definite diagnosis ofβ-KTD was made.Symptomatic treatment was adopted.Ten days later,the dyspnea was improved evidently and the ventilator was removed,but there were still obvious abnormalities on magnetic resonance imaging(MRI).The lesions mainly invaded the corpus striatum but were not limited to the basal ganglia.Then,the patient’s disease improved and discharged approximately 1 mo later,and the abnormal lesions on MRI had partially improved.However,for about 1 year,the residual irreversible lesions were observed on MRI,the mental and physical development of the patient was obviously regressive,and extra rehabilitation training was needed.CONCLUSION The case highlights the critical importance of one view that the range of lesions in some patients may be more extensive than previously thought in someβ-KTD patients.In addition to biochemical tests,genetic tests and magnetic resonance imaging are not only conducive to quickly diagnosingβ-KTD but also to partially evaluating the short-and long-term outcomes.Moreover,more attention should be paid to the two mutations(c.478 C>G;c.951 C>T)that may be associated with severeβ-KTD.

Effect of Yiqi Jianpi plus anticancer herbs on spleen deficiency in colorectal cancer and its anti-tumor role

Objective:To observe the effect of Yiqi Jianpi plus anticancer herbs on spleen deficiency in colorectal cancer and its anti-tumor role.Methods:Human intestinal cancer cell HT29 xenograft of nude mice model was established.The expression of ECF,VEGF,gastric cancer tumor growth in mice were observed.Results:Protein kinase C expression in in the Yiqi Jianpi group and Yiqi Jianpi anti-tumor group was significantly better than the model group(P<0.01,P<0.05).There was significantly more apoptotic cells in Yiqi Jianpi anti-tumor group than Yiqi Jianpi group and model group(P<0.01).Epidermal growth factor and vascular endothelial growth factor expression in Yiqi Jianpi group was significantly lower than Yiqi Jianpi group and model group(P<0.05).Conclusions:Tumor can inhibit the expression of PKC inhibition.Yiqi Jianpi and anticancer treatment can reduce this inhibition.Besides this treatment can also inhibit expression of tumor related genes such as epidermal growth factor and vascular endothelial growth factor.

The role of XPC protein deficiency in tobacco smoke-induced DNA hypermethylation of tumor suppressor genes

DNA hypermethylation of tumor suppressor genes has been frequently observed in cancer patients, and therefore, may provide a valuable biomarker for cancer prevention and treatment. DNA hypermethylation may also provide an important mechanism in cancer progression. Lung cancer is strongly associated with exposure to environmental carcinogens, especially tobacco smoke. DNA damage generated by tobacco smoke is believed to play an important role in lung cancer development. XPC is a DNA damage recognition protein required for DNA repair and other DNA damage responses and attenuated XPC protein levels have been detected in many lung cancer patients. We studied the role of XPC protein deficiency in tobacco smoke-caused DNA hypermethylation of important tumor suppressor genes. Using both normal human fibroblasts (NF) and XPC-deficient hu man fibroblasts (XPC), our DNA methylation studies demonstrated that the XPC deficiency caused elevated levels of DNA hypermethylation in both Brca1 and Mlh1 tumor suppressor genes following exposure to tobacco smoke condensate (TSC). The results of our ChIP studies revealed that the XPC deficiency led to an increased binding of DNA methyltransferase 3A (DNMT3A) at the promoter region CpG island-containing sequences of these genes under the TSC treatment;however, this increase was partially diminished with prior treatment with caffeine. The results of our immuno-precipitation (IP) studies demonstrated a protein-protein interaction of the ATR with DNMT3A. Our western blots revealed that the XPC deficiency caused an increase in TSC-induced ATR phosphorylation at serine 428, an indicator of ATR activation. All these results suggest that XPC deficiency causes an accelerated DNA hypermethylation in important tumor suppressor genes under tobacco smoke exposure and activation of the ATR signaling pathway is involved in this DNA hypermethylation process.

Evaluation of 30 DNA damage response and 6 mismatch repair gene mutations as biomarkers for immunotherapy outcomes across multiple solid tumor types

Objective:DNA damage response(DDR)genes have low mutation rates,which may restrict their clinical applications in predicting the outcomes of immune checkpoint inhibitor(ICI)treatment.Thus,a systemic analysis of multiple DDR genes is needed to identify potential biomarkers of ICI efficacy.Methods:A total of 39,631 patients with mutation data were selected from the cBioPortal database.A total of 155 patients with mutation data were obtained from the Fudan University Shanghai Cancer Center(FUSCC).A total of 1,660 patients from the MSK-IMPACT cohort who underwent ICI treatment were selected for survival analysis.A total of 249 patients who underwent ICI treatment from the Dana-Farber Cancer Institute(DFCI)cohort were obtained from a published dataset.The Cancer Genome Atlas(TCGA)level 3 RNA-Seq version 2 RSEM data for gastric cancer were downloaded from cBioPortal.Results:Six MMR and 30 DDR genes were included in this study.Six MMR and 20 DDR gene mutations were found to predict the therapeutic efficacy of ICI,and most of them predicted the therapeutic efficacy of ICI,in a manner dependent on TMB,except for 4 combined DDR gene mutations,which were associated with the therapeutic efficacy of ICI independently of the TMB.Single MMR/DDR genes showed low mutation rates;however,the mutation rate of all the MMR/DDR genes associated with the therapeutic efficacy of ICI was relatively high,reaching 10%–30%in several cancer types.Conclusions:Coanalysis of multiple MMR/DDR mutations aids in selecting patients who are potential candidates for immunotherapy.

RAD6B participates in neuronal DNA damage response to resist neurodegeneration

Protein ubiquitination plays an important role in DNA damage repair.To clarify the role of RAD6B,identified as an important ubiquitin conjugating enzyme,in neuronal genome integrity and survival,we first silenced RAD6B gene in RPE1 and found that RAD6B deficiency resulted in increased genomic instability.While in brains of RAD6B knockout mouse generated by homologous recom bination,we found that RAD6B deficiency caused a significant reduction of H2B ubiquitination and impaird DNA damage repair compared with wild type mouse brains.The accumulation of unrepaired DNA damage often leads to apoptosis or senescence,even tumor formation.Furthermore,we evaluated the effect of RAD6B deficiency on the survival of neurons,and noticed that RAD6B-deficient mice exhibited marked neuronal senescence,neurodegeneration and reduced neuronal numbers.We also observed learning and memory impairment in neuron-specific RAD6B-deficient mice.In conclusion,RAD6B is involved in DNA damage repair of neurons,and is essential to maintain neuronal genome stability.

从“虚瘀劫肝”论原发性肝癌中医药防治思路

理论创新是中医药发展的源泉,在新理论指导下形成的新方药或新诊治方案是中医药临床防治重大疑难疾病的新突破口。正邪交争,正不胜邪是慢性疾病持续进展的内在驱力和一般特性,慢性肝病持续恶性进展囊括了:肝纤维、肝硬化、肝癌等逐步恶化的病理进程。其在遵循中医“正不胜邪”基本病机的基础也必须有其特殊性和指向性,也可理解为在慢性肝病持续进展的病理状态下将一般性的病机进行指向性和特征性的阐发。肝正虚主要是指维持正常肝再生修复的相关机制和途径异常或紊乱,肝邪胜则是指肝损伤因素持续存在及其下游触发的恶性肝再生状态进行性亢进;而“瘀”作为正邪交争过程中产生的最主要病理产物也在不断推动疾病的恶性发展。在此基础上,从“虚瘀劫肝”论肝癌的病理新认识及补虚(补肾生髓、健脾)祛瘀(疏肝、柔肝、化肝)阻断慢性肝病持续恶性进展以防治肝癌的基本思路,期望有益于疑难肝脏病症的临床防治。

从“中焦如衡”论治胃癌前病变

“中焦如衡”是清代医家吴鞠通基于脾胃的生理特点,提出的脾胃病治疗大法。后世医家在此论点基础上,结合大量临床实践不断深化,形成了以“中焦如衡”为切入点的论治脾胃病病因病机与治疗大法为一体的理论体系。胃癌前病变(PLGC)是具有较高癌变风险的病理改变。从“中焦如衡”理论探讨PLGC的病因病机,立足于脾胃的生理病理特点,以“中焦如衡,以平为期”为切入点,认为本病由多种因素叠加导致胃黏膜遭受持续损害,而在正邪交争中,胃黏膜的保护和防御机制被激活,动态平衡机制被打破,出现以脾胃虚损为本、毒瘀损络为要、升降失衡为主要表现的病机特点,随之而出现病理改变和临床表现。通过调和气血、虚实、气机等多种因素,以调和中焦为手段,以恢复脾胃平衡、气机升降为目的,从而防治PLGC。

基于“中气下陷”理论探析膜性肾病的病机及诊治思路

基于中气下陷理论探讨膜性肾病的病机,提出“湿蕴脾肾,中气下陷”的病机理论,其中以风邪外侵,中土亏损为发病先导,中气不足,脾肾亏虚为发病基础,气机凝滞,湿瘀阻络为病机核心,毒损脾肾,中气下陷为病情进展,大气虚散,脏腑衰惫为病重结局,中气亏虚贯穿膜性肾病发病始终。论治当取升阳举陷大法,强调早期调补脾肾,御风于外,中期祛湿化瘀解毒,升阳通络,晚期升阳益胃排毒,兼顾心肾虚损。

基于“伏邪理论”探讨中医药抗溃疡性结肠炎复发的诊疗策略

溃疡性结肠炎(ulcerative colitis,UC)是一种病因尚未阐明的好发于直肠和结肠的慢性炎症性疾病,目前尚无确切有效的治疗方法,其具有周期性、反复发作等发病特点,此与中医学“伏邪理论”认识趋同。该文试从中医“伏邪理论”入手,回顾相关典籍,结合本团队的临床经验,对UC的中医发病机制进行重新探讨。认为湿热、瘀血是伏邪的重要组成部分,脾虚是伏邪产生的基础,而伏邪所致的肠络受损是UC发生的病理基础,因此临证遣方用药时谨遵扶正祛邪、透邪外出的治疗总纲,提出清热利湿是祛除伏邪的首要治则,健脾温中为重要保障,理气活血是主要方式,而顾护胃气则需贯穿治疗始终,由此创立的清肠温中方具有明显的临床疗效,并能有效防治UC的复发。该文分别从“伏邪理论”源流、伏邪在UC发病中的特点及在诊疗中的应用等探讨了中医药抗UC复发的诊疗策略,并附本团队防治UC复发的经验,以期对该病的临床治疗有所裨益。

《黄帝内经》“血脉”整体观防治动脉粥样硬化性心血管疾病的理论发挥

《黄帝内经》有云:“人之所以成生者,血脉也”“血脉和利,精神乃居”“血”,维系着机体的正常生理功能;“脉”,则具有推动和约束血液在脉道运行的作用;“血”与“脉”相互作用,二者相合,共同维持着机体的生命活动。传承《黄帝内经》的思想和理论,杨关林教授从中医学“整体观”出发,结合前人“痰瘀”相关学说,整体审视心脑血管系统疾病,提纲挈领地提出“血脉病”这一概念。血脉病,指气血运行失常,导致血脉痹阻的一类疾病,包含心脉病、脑脉病和肢脉病等多系统疾病。血脉病的发生与脾虚受损有密切的联系,我们创新性的将血脉病分为“血浊脉滞”“血瘀脉壅”“血毒脉损”三个不同发展阶段,并根据不同阶段的病机和临床表现提出“健脾益气,化痰降浊”“健脾益气,化痰祛瘀”“化痰祛瘀,清热解毒”三种不同的治法并建立出相应中药组方,构建血脉病全程防治体系,提升中医药防治疗效。

基于“心脑同治”理论研究佛手养心汤改善气虚血瘀型心衰后认知障碍模型大鼠心肌线粒体损伤的作用

目的:基于“心脑同治”理论探索佛手养心汤对气虚血瘀型心衰后认知障碍模型大鼠的影响,及对心肌线粒体结构与能量代谢损伤的改善作用。方法:通过体内实验,将48只SD大鼠随机选取10只作为正常组,剩余大鼠予以异丙肾上腺素腹腔注射、左旋硝基精氨酸灌胃以建立心衰后认知障碍模型,然后采取饥饿法和力竭游泳法以建立气虚血瘀型模型。将造模成功后的大鼠随机分为模型组、治疗组和对照组,每组10只。正常组和模型组大鼠常规饲养,治疗组予以佛手养心汤干预,对照组予以盐酸曲美他嗪片干预,为期4周。观察大鼠的一般情况,运用动物心脏超声仪检测大鼠心脏LVESD、LVEDD、LVEF指标;Morris水迷宫检测认知功能;透射电镜下观察心肌线粒体结构改变;比色法检测心肌线粒体mPTP开放程度;微量法检测心肌组织ATP含量。结果:造模成功后大鼠一般情况差,反应迟钝,皮肤和毛发枯燥无光泽,活动减少,进食缓慢,饮水量减少,唇甲发绀,部分大鼠出现气促而喘的症状。造模结束予以药物干预后,大鼠上述表现可明显改善。心动超声检测提示模型组大鼠LVESD、LVEDD明显增厚,LVEF明显降低,而治疗组、对照组可显著逆转上述变化。Morris水迷宫检测显示模型组大鼠逃避潜伏期明显延长,且穿越平台次数明显减少,而治疗组与对照组检测结果与之相反。与此同时,以透射电镜观察线粒体结构、比色法与微量法检测线粒体能量代谢状态,发现模型组大鼠线粒体结构损伤明显,线粒体mPTP开放程度显著增加,心肌组织ATP含量明显降低,而治疗组与对照组给予相应药物干预后可显著逆转上述改变。结论:佛手养心汤可有效抑制心衰大鼠心肌线粒体mPTP开放,改善线粒体结构损伤,增加心肌组织ATP含量,逆转心功能改变,从而改善气虚血瘀型心衰后认知障碍大鼠的线粒体功能和认知功能。

赵晓平基于“虚瘀毒损”治疗脑胶质瘤术后复发经验

总结赵晓平对脑胶质瘤术后复发患者的中医病机认识及其临证诊治经验。赵晓平基于“虚瘀毒损”理论提出脑胶质瘤术后复发的核心病机为“气虚毒染以致瘀”,认为益气祛瘀解毒为本病的根本治法,并运用益气祛瘀解毒方治疗脑胶质瘤术后患者。选取赵晓平既往临床预防脑胶质瘤术后复发的一例经典临证医案,详细分析案例中患者病情转归过程的各阶段用药思路,对赵晓平运用益气祛瘀解毒法治疗脑胶质瘤术后复发经验进行分析总结,为中医药防治脑胶质瘤复发提供借鉴和参考。

加味黄精四草汤辅治肝肾阴虚型高血压早期肾损害的临床效果

目的 观察加味黄精四草汤辅治肝肾阴虚型高血压早期肾损害的临床效果。方法 选取2019年12月—2021年12月济南市中西医结合医院收治的肝肾阴虚型高血压早期肾损害患者110例,按随机数字表法分为研究组55例和对照组55例,因在治疗中有脱落情况,最终研究组48例,对照组48例。对照组予常规治疗,研究组在对照组基础上加用加味黄精四草汤治疗,2组均治疗2个月。比较2组临床疗效,治疗前后血压、血清β2微球蛋白(β2-MG)、尿微量白蛋白(mAlb)、血清胱抑素C(CysC)、肝肾功能指标[尿素、肌酐、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)],患者满意度及不良反应。结果 研究组总有效率为97.92%,高于对照组的81.25%(χ^(2)=7.144,P=0.008)。治疗2个月后,2组收缩压、舒张压及血清β2-MG、mAlb、血清CysC水平低于治疗前,且研究组低于对照组(P<0.01);治疗前后,2组尿素、肌酐、ALT、AST水平比较差异均无统计学意义(P>0.05)。研究组患者总满意度为95.83%,高于对照组的72.92%(χ^(2)=9.561,P=0.002);研究组与对照组不良反应总发生率比较差异无统计学意义(0 vs. 6.25%,χ^(2)=1.376,P=0.241)。结论 加味黄精四草汤辅治肝肾阴虚型高血压早期肾损害疗效显著,可有效保护患者肝肾功能,稳定患者血压,降低血清β2-MG、mAlb、CysC水平,提高患者满意度,安全性高。

《伤寒论》身重病因病机

《伤寒论》中的身重病机有风寒外束,里热炽盛,气机郁滞,水湿不化,阳气受损,精气亏虚等,其最后转归均为经脉受邪,失于濡养。风寒外束,阳气闭塞,寒性收引,表闭不开,阳郁不伸,四肢经脉失养,则发为身重,用大青龙汤解表清热;阳明里热炽盛,伤津耗气,周身经脉失于濡养则身重,用大承气汤破实攻满,或用白虎汤清阳明之热;少阳枢机不利,气机失于条达,运行不畅,经脉失养,发为身重,用柴胡加龙骨牡蛎汤疏通三焦气机,驱阳达表;少阴病肾阳虚损,水湿不化,浸于四肢,导致四肢沉重疼痛,用真武汤温阳利水;表邪不解误用灸法,表闭阳郁,火性炎上,上实下虚,故见下肢沉重麻木,用肉桂、附子等温通下焦阳气,龙骨、牡蛎等潜镇在上之浮阳,表邪不解误用下法,阳气受损,经脉失养,可见身体困重,治疗当以补益扶正为主,方用小建中汤或补中益气汤;伤寒后正气亏虚,行房事更耗伤精气,四肢失于濡养,形气两伤,故身重,治疗应以扶正为主,方用烧裩散。

Study on Relation of Kidney-Deficiency with Hearing Damage and Serum Trace Elements in Patients with Chronic Nephropathy

Objective: To find some objective criteria for Syndrome Differentiation of Kidney Deficiency by analysing the relationship between Kidney and hearing as well as serum trace elements. Methods: The hearing and serum trace element of 73 cases of chronic nephropathy were measured. Results: The incidence of hearing damage in high frequency was 68. 5% with a mean hypoacusis of 35. 1 ±6. 1 dB, while in the 55 patients of Kidney deficiency type, the incidence of hearing damage in high frequency was 83. 6%, which was significantly higher than that in patients without Kidney Deficiency. The serum levels of trace elements (iron and zinc) in patients with Kidney Deficiency were lower than the normal level and that in patients without Kidney Deficiency, and the decrease of serum iron was directly proPOrtional to the degree of hearing damage. Conclusion: There was close relationship between Kidney Deficiency and hearing damage as well as the serum levels of iron and zinc.

椎基底动脉扩张延长症病机的中医“虚气留滞”病机假说阐释

椎基底动脉扩张延长症是一种预后不良的后循环血管变异疾病,目前尚缺乏有效的防治手段。通过对其病理机制、临床特征与转归的总结分析,与中医“虚气留滞”理论相结合,提出了创新病机假说,以“虚气”为发病之本,以“留滞”为发病之标,二者具有互为因果、相互促进的特点,切合疾病螺旋进展、动态演变的模式。并据此形成“扶正护脑以补虚、解毒通络而通滞”的治法,为进一步探索椎基底动脉扩张延长症的中医证治规律与疗效评价方法,为临床实践提供依据,对提高临床疗效,降低其相关的神经系统疾病负担具有重要意义。

基于“脾病下流乘肾”探讨糖尿病肾病研究进展

糖尿病肾病(DKD)因其发病因素、所处病理阶段及临床表现的错综复杂性,致使诸多医家无法准确把握疾病的病因病机及治法。通过对近5年的中英文相关文献进行梳理与总结,结合DKD分为隐性期、显性期以及终末期的特点,以“脾病下流乘肾”为纲,从气阴两虚、脾肾阳虚、毒损肾络等三方面详细论述了DKD的病因病机及治法,以期为中医药防治DKD提供新的思路与方法。

从扶正祛邪论治颈动脉粥样硬化斑块溃疡

动脉粥样硬化是心脑血管疾病的始动罪犯因素,严重威胁着人类生命健康,易损斑块易破溃或形成血栓是致病和诱发不良事件的高危因子。斑块的稳定与否影响并决定着动脉硬化的发展方向及进程,易损斑块的稳定性受巨噬细胞、多种蛋白酶学分子、细胞凋亡、血管新生、老化及生物力学等多因素影响。现代医学对动脉硬化斑块的防治方式包括以药物为主的内科治疗和以手术或介入的外科治疗,能起到一定防治作用但也伴随着不良反应和复发率的增加。传统医学多从瘀、痰、毒、虚论治动脉粥样硬化及斑块的易损性,多强调邪实的重要性而不注重正虚或两者兼顾的情况,作者认为动脉硬化斑块的形成、受损、不稳定、破溃、出血等病理特征病机的本质为正虚毒损,正虚与邪实两者兼而有之。正虚为血气不和或不足,营卫失调,正虚失固,邪实为痰、瘀、毒诸邪壅盛侵袭,机体损伤致病,治当扶正祛邪,虚实兼顾,化瘀祛痰,生肌稳斑。

张发荣教授补肾和络法治疗糖尿病周围神经病变经验

[目的]总结全国名中医张发荣教授治疗糖尿病周围神经病变的临证经验。[方法]通过跟师侍诊,整理医案及相关文献,从病因病机、处方用药等方面总结张教授治疗糖尿病周围神经病变的经验,并附验案一则。[结果]张教授结合多年临证经验,认为糖尿病周围神经病变以“肾虚络损”为基本病机,遵循以治疗糖尿病本病为经,治疗周围神经病变为纬的“经纬论治”法,以补肾和络为基本治则,补肾善从“阴中求阳”“阳中求阴”;和络则灵活运用养阴清热宁络、益气养阴行络、健脾化湿通络、豁痰化瘀畅络、清热利湿通络、温经散寒护络等方法。临证中尤其重视虫类药物的使用,以活血通络,同时重调脾胃,不避甘温,内外合治,双管齐下。所举验案辨证为脾肾亏虚、络脉瘀阻证,张教授治以补肾活血通络,兼以健脾祛湿,临床疗效显著。[结论]张老把握糖尿病周围神经病变肾虚络损的基本病机,采用“经纬论治”法,治疗重在补肾和络,临床疗效显著,值得学习借鉴。