Guideline for clinical comprehensive evaluation of Chinese patent medicine (2022 version) / 中国中药杂志

Currently,the research or publications related to the clinical comprehensive evaluation of Chinese patent medicine are increasing,which attracts the broad attention of all circles. According to the completed clinical evaluation report on Chinese patent medicine,there are still practical problems and technical difficulties such as unclear responsibility of the evaluation organization,unclear evaluation subject,miscellaneous evaluation objects,and incomplete and nonstandard evaluation process. In terms of evaluation standards and specifications,there are different types of specifications or guidelines with different emphases issued by different academic groups or relevant institutions. The professional guideline is required to guide the standardized and efficient clinical comprehensive evaluation of Chinese patent medicine and further improve the authority and quality of evaluation. In combination with the characteristics of Chinese patent medicine and the latest research achievement at home and abroad,the detailed specifications were formulated from six aspects including design,theme selection,content and index,outcome,application and appraisal,and quality control. The guideline was developed based on the guideline development requirements of China Assoication of Chinese medicine. After several rounds of expert consensus and public consultation,the current version of the guideline has been developed.

Pirfenidone alleviates urethral stricture following urethral injury in rats by suppressing TGF-β1 signaling and inflammatory response / 南方医科大学学报

OBJECTIVE@#To investigate the effect of pirfenidone for reducing urethral stricture following urethral injury in rats and explore the possible mechanism.@*METHODS@#Thirty male SD rats were randomly assigned into negative control group, positive control group and pirfenidone group (n=10). In pirfenidone and positive control groups, the rats were subjected to incision of the posterior urethral cavernous body followed by daily intraperitoneal injection of pirfenidone (100 mg/kg) and an equivalent volume of solvent, respectively. The rats in the negative control group were given intraperitoneal injections of solvent without urethral injury. At two weeks after modeling, retrograde urethrography was performed for observing urethral stricture, and the injured urethral tissues were harvested for HE staining, Masson staining, immunohistochemical staining and Western blotting for detecting the protein expressions of α-SMA and TGF-β1. The mRNA expressions of the inflammatory factors TNF-α, IL-6, and IL-1β were detected using qRT-PCR.@*RESULTS@#The body weight of the rats in pirfenidone group was significantly decreased compared with that in the other two groups (P < 0.05). Retrograde urethrography showed significant narrowing of the urethra in the positive control group but not in the pirfenidone group. HE staining of the injured urethral tissues showed obvious proliferation of urethral epithelial cells with narrow urethral cavity and increased inflammatory cells in positive control group. The pathological findings of the urethra were similar between pirfenidone group and the negative control group. Masson staining revealed obviously reduced collagen fibers and regular arrangement of the fibers in pirfenidone group as compared to the positive control group. Compared with those in the negative control group, the expressions of α-SMA and TGF-β1 were significantly increased in the positive control group, and pirfenidone treatment significantly inhibited their expressions (P < 0.05 or 0.01). Pirfenidone also significantly inhibited the mRNA expressions of TNF-α, IL-6, and IL-1β in the injured urethral tissue (P < 0.05 or 0.01).@*CONCLUSION@#Pirfenidone can prevent urethral fibrosis and stricture after urethral injury possibly by inhibiting the TGF-β1 pathway and inflammatory response.

Centromere protein U enhances the progression of bladder cancer by promoting mitochondrial ribosomal protein s28 expression

Bladder cancer is one of the most common types of cancer. Most gene mutations related to bladder cancer are dominantly acquired gene mutations and are not inherited. Previous comparative transcriptome analysis of urinary bladder cancer and control samples has revealed a set of genes that may play a role in tumor progression. Here we set out to investigate further the expression of two candidate genes, centromere protein U (CENPU) and mitochondrial ribosomal protein s28 (MRPS28) to better understand their role in bladder cancer pathogenesis. Our results confirmed that CENPU is up-regulated in human bladder cancer tissues at mRNA and protein levels. Gain-of-function and loss-of-function studies in T24 human urinary bladder cancer cell line revealed a hierarchical relationship between CENPU and MRPS28 in the regulation of cell viability, migration and invasion activity. CENPU expression was also up-regulated in in vivo nude mice xenograft model of bladder cancer and mice overexpressing CENPU had significantly higher tumor volume. In summary, our findings identify CENPU and MRPS28 in the molecular pathogenesis of bladder cancer and suggest that CENPU enhances the progression of bladder cancer by promoting MRPS28 expression.

Centromere protein U enhances the progression of bladder cancer by promoting mitochondrial ribosomal protein s28 expression

Bladder cancer is one of the most common types of cancer. Most gene mutations related to bladder cancer are dominantly acquired gene mutations and are not inherited. Previous comparative transcriptome analysis of urinary bladder cancer and control samples has revealed a set of genes that may play a role in tumor progression. Here we set out to investigate further the expression of two candidate genes, centromere protein U (CENPU) and mitochondrial ribosomal protein s28 (MRPS28) to better understand their role in bladder cancer pathogenesis. Our results confirmed that CENPU is up-regulated in human bladder cancer tissues at mRNA and protein levels. Gain-of-function and loss-of-function studies in T24 human urinary bladder cancer cell line revealed a hierarchical relationship between CENPU and MRPS28 in the regulation of cell viability, migration and invasion activity. CENPU expression was also up-regulated in in vivo nude mice xenograft model of bladder cancer and mice overexpressing CENPU had significantly higher tumor volume. In summary, our findings identify CENPU and MRPS28 in the molecular pathogenesis of bladder cancer and suggest that CENPU enhances the progression of bladder cancer by promoting MRPS28 expression.

Effect of Sijunzi Decoction on regulation of Aβ-related proteins across blood-brain barrier / 中国中药杂志

According to traditional Chinese medicine, "spleen transport" is closely related to the metabolism of substance and energy. Studies have shown that Alzheimer's disease(AD) is a disease related to glucose and lipid metabolism and energy metabolism. The traditional Chinese medicine Jiangpi Recipe can improve the learning ability and memory of AD animal model. Sijunzi Decoction originated from Taiping Huimin Hefang Prescription is the basic prescription for strengthening and nourishing the spleen, with the effects of nourishing Qi and strengthening the spleen. In this experiment, human brain microvascular endothelial cells(HBMEC) and Sijunzi Decoction water extract(0.25, 0.5, 1 mg·L~(-1)) were pre-incubated for 2 h, and then Aβ_(25-35) oligomers(final concentration 40 μmol·L~(-1)) was added for co-culture for 22 hours. The effect of Sijunzi Decoction on the activity of Aβ_(25-35) oligomer injured cells and the expression of related proteins were investigated. Q-TOF-LC-MS was used first for principal component analysis of Sijunzi Decoction water extract. Then MTT assay was used to investigate the effect of Sijunzi Decoction water extract on the proliferation of HBMEC cells. Real-time fluorescence quantitative PCR(RT-qPCR) was employed to detect the mRNA expression of GLUT1, RAGE, and LRP1. The expression of Aβ-related proteins across blood-brain barrier(RAGE, LRP1) was detected by Western blot. The results showed that 40 μmol·L~(-1) Aβ_(25-35) oligomers could induce endothelial cell damage, reduce cell survival, increase expression of RAGE mRNA and RAGE protein, and reduce expression of GLUT1 mRNA, LRP1 mRNA, and LRP1 protein. Sijunzi Decoction water extract could reduce the Aβ_(25-35) oligomer-induced cytotoxicity of HBMEC, decrease the expression of RAGE mRNA and RAGE protein, and increase the expression of GLUT1 mRNA, LRP1 mRNA and LRP1 protein. The results indicated that Sijunzi Decoction could reduce the injury of HBMEC cells induced by Aβ_(25-35) oligomer, and regulate the transport-related proteins GLUT1, RAGE and LRP1, which might be the mechanism of regulating Aβ_(25-35) transport across the blood-brain barrier.

Investigation of Relationship Between Spleen and Alzheimer' s Disease Based on Glucose, Lipid and Energy Metabolisms / 中国实验方剂学杂志

Alzheimer' s disease (AD) is a chronic neurodegenerative disease, which is seriously affecting people' s lives. Until now, the pathogenesis of AD is still unknown which result in its worldwide difficulties in prevention and treatment. Some studies have shown that AD might be a metabolic disease associated with glucose, lipid and energy metabolisms. Traditional Chinese medicine (TCM) believes that the spleen is acquired foundation and the origination of Qi and blood. The function of spleen is not only closely related to the metabolism of substance and energy, but also related to the aging of human body. In this article, we summarized and analyzed the interrelationship of metabolism, AD and spleen, as well as the effect of spleen-invigorating prescription on AD. The purpose of the paper is to analyze the possible mechanism of TCM treating AD from the perspective of regulating metabolism, explore the potential value of spleen-strengthening TCM in the treatment or prevention of AD, and provide new ideas for exploring the drug development and TCM therapy of AD.

Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ25-35 / 中国实验方剂学杂志

Objective::To observe the neuroprotective effect and potential mechanism of Zhenxin Shengshui Yizhi Fang(XSF) aqueous extract on human brain microvascular endothelial cells (HBMEC) injury induced by amyloid-β protein(Aβ)25-35. Method::HBMEC cells damage induced by Aβ25-35 was used as Alzheimer' s disease(AD) cell model. The study included control group, Aβ25-35 group, and low, medium and high-dose XSF aqueous extract groups (125, 250, 500 mg·L-1). After treatment, the cytotoxicity of different concentrations of drugs and Aβ25-35 was determined by methyl thiazolyl tetrazolium(MTT) colorimetry. Apoptosis was observed by Hoechst-33258 staining. The activity of Caspase-3 was detected by colorimetry. Western blot was used to detect the expression levels of the receptor of advanced glycation end products (RAGE) and low-density lipoprotein receptor-related protein (LRP1). Result::Compared with the control group, the cell viability of Aβ25-35 group was significantly decreased (P<0.01). Hoechst-33258 staining showed bright blue fluorescence, chromatin condensation, dense staining or fragmentation dense staining, whitening in color, and significant increase of the percentage of apoptotic cells (P<0.01). Caspase-3 activity increased significantly (P<0.01). Western blot showed that RAGE protein expression increased significantly (P<0.01), while low-density lipoprotein receptor-related protein(LRP1), glucose transporter 1(GLUT1) and GLUT3 protein expressions decreased significantly (P<0.01). Compared with the Aβ25-35 group, the cell viability of XSF aqueous extract groups was significantly increased in a dose-dependent manner. The XSF aqueous extract had a more significant protective effect of than the other groups (P<0.05). The XSF aqueous extract group (500 mg·L-1) significantly inhibited the number of apoptotic cells (P<0.01), but significantly reduced the Caspase-3 activity (P<0.01). RAGE protein expression was not significantly decreased in XSF aqueous extract group (125 mg·L-1), but significantly decreased in XSF aqueous extract group (250, 500 mg·L-1, P<0.01), while LRP1, GLUT1 and GLUT3 protein expression significantly increased (P<0.05, P<0.01) in a dose-dependent manner. Conclusion::XSF aqueous extract can attenuate the cytotoxicity of HBMEC induced by Aβ25-35 oligomer, inhibit apoptosis, decrease caspase-3 activity and RAGE protein expression, increase LRP1, GLUT1 and GLUT3 protein expressions, and reduce the abnormal accumulation and deposition of Aβ in the brain, which may be its mechanisms for prevention and treatment of AD.

Effect of Zhenxin Xingshui Yizhi Fang and Its Essential Oil Extract on Ability of Learning and Memory in Alzheimer's Disease Model Mice / 中国实验方剂学杂志

Objective: The present study was designed to investigate the modulating effect of Zhenxin Xingshui Yizhi Fang and its essential oil extract on cognitive deficits in mice.Method: For the purpose of this study 5 months old APP/PS1 double transgenic mice and wild-type C57BL/6JNju were selected as experimental animals.Then APP/PS1 double transgenic mice were randomly divided into model group,essential oil low and high-dose groups (12.13,48.50 mg·L-1),Zhenxin Xingshui Yizhi Fang group (0.46 g·kg-1).Meanwhile,wild-type C57BL/6JNju mice were used as a normal group.APP/PS1 double transgenic mice were treated with Zhenxin Xingshui Yizhi Fang and its essential oil extract for 22 consecutive days.Mice were subjected to a Morris water maze test and a platform test in order to determine their cognitive effect.Nissl's staining was used to observe pathological changes in brain tissue.Meanwhile,senile plaques (SP) were observed by employing Thioflavin-S staining.The expression of glucose transporter 1(GLUT1) and insulin receptor substrate-1(IRS-1) were analyzed using immunohistochemistry techniques.The levels of neurotransmitters such as acetylcholine (ACH),glutamate (GLU) and γ-aminobutyric acid (GABA) in the hippocampus were quantified by enzyme-linked immunosorbent assay (ELISA).Result: The memory function was significantly reduced in model group,and severe brain injury and neuronal apoptosis were also observed in comparison to normal group (PPPPPPPConclusion: These results indicate that Zhenxin Xingshui Yizhi Fang and its essential oil extract could ameliorate cognitive deficits and GLUT1 and IRS-1 could be a possible therapeutic target for AD.It may be an interesting approach to the treatment of Alzheimer's disease.

Effects of hydrogen sulfide (HS) on cardiac hypertrophy and miRNA-133a-mediated Ca/calcineurin/NFATc4 signal pathway in rats / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of hydrogen sulfide (HS) on the negatively regulation of cardiomyocyte hypertrophy and the relationship between the effect of HS with miRNA-133a-mediated Ca/calcineurin/NFATc4 signal pathway.@*METHODS@#Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (Leica). The expression of brain natriuretic peptide(BNP), β-myosin heavy chain(β-MHC), cystathionase (CSE), miRNA-133a, calcineurin (CaN) were detected by qRT-PCR. The protein expressions of CaN、nuclear factors of activated T cells (NFATc4) were detected by Western blot. The concentration of HS in the cardiomyocyte was detected by Elisa. The concentration of intracellular calcium was measured by calcium imaging using confocal microscope. The nuclear translocation of NFATc4 was checked by immuno-fluorescence cell staining technique.@*RESULTS@#①The level of system of CSE/HS and expression of miRNA-133a were significantly reduced in cardiomyocyte hypertrophy. Pretreatment with NaHS increased the concentration of HS and the expression of miRNA-133a mRNA in cardiomyocytes, and suppressed cardiomyocyte hypertrophy. ②The concentration of intracellular calcium, the expression of CaN and nulear protein NFATc4 were significantly increased, and the nuclear translocation of NFATc4 were obviously enhanced in cardiomyocyte hypertrophy. NaHS pretreatment markedly inhibited these effects of ISO induced cardiomyocyte hypertrophy. ③Application of antagomir-133a reversed the inhibitory effects of NaHS on cardiomyocyte hypertrophy, and increased the influx of intracellular calcium, and elevated the expression of CaN and nuclear protein NFATc4, and enhanced the nuclear translocation of NFATc4.@*CONCLUSIONS@#HS can negatively regulate cardiomyocyte hypertrophy. The effects might be associated with HS increasing expression of miRNA-133a and inhibiting inactivation of Ca/calcineurin/NFATc4 signal pathway.

Ophiopogonin-B inhibited cell mitosis by Myt / Cdc2 pathway in H460 cells / 中国药理学通报

Aim To explore the effects of ophiopogo-nin-B (OP-B)on cell cycle and mitosis in non-small cell lung cancer (NSCLC)H460 cells in vitro and the underlying mechanisms. Methods TUNEL immuno-histochemical assay was used to detect the change of the nuclear matter. DAPI staining was used to detect the change of the nuclear morphology and the mitosis status. Meanwhile,Western blot was performed to de-termine the protein level of the proteins regulating cell cycle and mitosis. Results OP-B significantly arres-ted cell cycle in G0 / G1 phase and inhibited the mitosis in H460 cells at the concentration of 10 μmol·L - 1 . Meanwhile,it inhibited the protein level of cyclinD1, cyclinB1,and up-regulated the expression of Myt and the phosphorylation level of Cdc2. Conclusion OP-B inhibits cell mitosis in A549 cells through Myt/ Cdc2 signaling pathway.

The effect of relgulation of PPAR-α on cardiac hypertrophy and the relationship between the effect of PPAR-α with PI3K/Akt/mTOR pathway / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of peroxisiome proliferator activated receptor-α (PPAR-α) on the regulation of cardiomyocyte hypertrophy and the relationship between the effect of PPAR-α with PI3K/Akt//mTOR signal pathway.</p><p><b>METHODS</b>Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (Leica). The expressions of atrial natriuretic peptide (ANP), β-myosin heavy chain (β-MHC) and PPAR-α mRNA were detected by qRT-PCR. The protein expressions of Akt, mTOR and P70S6K were detected by Western blot. The expression of PPAR-α was suppressed by RNAi.</p><p><b>RESULTS</b>(1) The expression of PPAR-α was significantly reduced in cardiomyocyte hypertrophy. PPAR-α activator Fenofibrate (Feno) increased the expression of PPAR-α and suppressed cardiomyocyte hypertrophy. The inhibitory effect of Feno on cardiomyocyte hypertrophy was reversed by PPAR-α RNAi. (2) Feno significantly inhibited the increase of the protein expressions of p-Akt, p-mTOR and p-p70S6K in ISO induced cardiomyocyte hypertrophy, which could be blocked by PPAR-α RNAi. (3) PI3K antagonist LY294002 (LY) or mTOR antagonist rapamycin (RAPA) markedly-inhibited cardiomyocyte hypertrophy. The inhibitory effects of LY or RAPA on cardiomyocyte hypertrophy were reversed by PPAR-α RNAi.</p><p><b>CONCLUSION</b>PPAR-α can negatively regulate cardiomyocyte hypertrophy. The effect might be associated with PPAR-α inhiting PI3K/ Akt/mTOR signal pathway.</p>

Hyperactivation of c-Jun NH(2)-terminal protein kinase contributes to the proliferation of B lymphoma cells / 中国实验血液学杂志

This study was purposed to explore the effect of hyperactivation of c-Jun NH(2)-terminal protein kinase (JNK) on the proliferation of B lymphoma cells. The human B lymphoma cell lines Daudi and Raji were chosen as research objects. The expression of JNK protein was determined by Western blot. The subcellular localization of JNK protein was detected by immunofluorescence. The cell cycle was analyzed by flow cytometry. The suppressive effect of JNK inhibitor SP600125 on the proliferation of Daudi and Raji cells was assayed by ATPLite method. The results demonstrated that hyperactivation of JNK has been found in Daudi and Raji cells. Immunofluorescence confirmed the aberrant subcellular localization of JNK protein in Daudi and Raji cells. Cell cycle assay revealed that Daudi and Raji cells underwent G(2)-M arrest in the presence of SP600125. Furthermore, Daudi and Raji cells showed significant increase in sub-G(1) population, an indicator of apoptotic cells, with the treatment of JNK inhibitors. These data suggested that JNK inhibitors suppressed the growth of B lymphoma cells via cell cycle arrest and apoptosis. Daudi and Raji cells treated with different concentrations of JNK selective inhibitor SP600125 showed dose-dependent reduction in the growth of Daudi and Raji cells. It is concluded that hyperactivation of JNK enhance the proliferation of Daudi and Raji cells. The aberrant subcellular localization of JNK protein may facilitate the nuclear accumulation of basal JNK activity, which made JNK to be a potential target to treat human B lymphoma.

Functional changes of intestinal mucosal barrier in surgically critical patients / 世界急诊医学杂志（英文）

BACKGROUND: The gut is capable of inducing multiple organ dysfunction syndrome (MODS). In the diagnosis and treatment of critical ill patients, doctors should pay particular attention to the protection or recovery of intestinal barrier function. However, no reliable diagnostic criteria are available clinically. This study aimed to assess the changes of intestinal mucosal barrier function in surgically critical ill patients as well as their significance. METHODS: Thirty-eight surgically critical ill patients were enrolled as a study group (APACHE II>8 scores), and 15 non-critical ill patients without intestinal dysfunction were selected as a control group (APACHE II<6). General information, symptoms, physical signs, and APACHE II scores of the patients were recorded. The patients in the study group were subdivided into an intestinal dysfunction group (n=26) and a non-intestinal dysfunction group (n=12). Three ml venous blood was collected from the control group on admission and the same volume of plasma was collected from the study group both on admission and in the period of recovery. The plasma concentrations of endotoxin, diamine oxidase (DAO), D-lactate, and intestinal fatty-acid binding protein (iFABP) were detected respectively. The data collected were analyzed by the SPSS 17.0 software for Windows. RESULTS: The levels of variables were significantly higher in the study group than in the control group (P<0.01). They were higher in the intestinal dysfunction group than in the non-intestinal dysfunction group (DAO P<0.05, endotoxin, D-lactate, iFABP P<0.01). In the non-intestinal dysfunction group compared with the control group, the level of endotoxin was not significant (P>0.05), but the levels of DAO, D-lactate and iFABP were statistically significant (P<0.05). The levels of variables in acute stage were higher than those in recovery stage (P<0.01).The death group showed higher levels of variables than the survival group (endotoxin and D-lactate P<0.01, DAO and iFABP P<0.05). CONCLUSION: The plasma concentrations of endotoxin, DAO, D-lactate, and intestinal fatty-acid binding protein (iFABP) could reflect a better function of the intestinal mucosa barrier in surgically critical ill patients.