Simultaneous determination of fifteen components in Niuhuang Jiangya Pills by UPLC-Q Exactive-Orbitrap MS / 药学学报

An ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q Exactive-Orbitrap MS) method for the simultaneous determination of 15 compounds (taurocholic acid, 7-keto-3α,12-α-dihydroxycholanic acid, glycocholic acid, 3-oxo-7α,12α-hydroxy-5β-cholanoic acid, taurochenodeoxycholic acid, 3α-hydroxy-7-oxo-5β-cholanic acid, hyocholic acid, sodium taurodeoxycholate, hyodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycodeoxycholic acid, taurolithocholic acid sodium salt, chenodeoxycholic acid, deoxycholic acid) in Niuhuang Jiangya Pills was established. The separation was performed on a Thermo Fisher Scientfic Bremen HYPERSIL GOLD C18 column (100 mm × 2.1 mm, 1.9 μm). Methanol and water (containing 0.1% formic acid)were adopted as the mobile phase by gradient elution．MS detection was performed with multiple reaction monitoring mode．The results showed that fifteen compounds had a good linearity within their respective concentration ranges (r > 0.999 0). The average recovery rates were 93.7%- 105.2% (n = 9). The established method was used to determine the content of 15 batches of samples, and the results showed that the content of cholic acid was quite different. The present study provides an important reference for the overall quality control of Niuhuang Jiangya Pills.

Total flavonoids of Dracocephalum moldavica L. inhibit oxidative stress against H9c2 cell ischemia-reperfusion injury via VEGF-B/AMPK pathway / 药学学报

Total flavonoids of Dracocephalum moldavica L. (TFDM) is an effective component extracted and isolated from the traditional Uighur medicinal herb Cymbidium fragrans. Cymbidium fragrans has the effects of tonifying the heart and brain, promoting blood circulation and resolving blood stasis, and has been widely used in the treatment of cardiovascular and cerebrovascular diseases for a long time. The purpose of this study was to determine the effect of total flavonoids from Cymbidium fragrans on hypoxia/re-oxygenation (H/R) injury in H9c2 (rat cardiomyocytes) cells and its mechanism. A model (H/R) of hypoxia/re-oxygenation injury in H9c2 cells was established using hypoxia and glucose deprivation for 9 h combined with re-oxygenation and rehydration for 2 h to simulate myocardial ischemia-reperfusion injury. The effects of total flavonoids from Cymbidium fragrans on cell viability, markers of myocardial cell damage, oxidative stress levels, and reactive oxygen radical (ROS) content were investigated, Western blot was used to detect the expression of vascular endothelial growth factor B (VEGF-B) and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) pathway related proteins. The results showed that the total flavonoids of Cymbidium fragrans significantly increased the viability of myocardial cells after H/R injury, and decreased the content of lactate dehydrogenase (LDH) and creatine kinase isozyme (CK-MB) in the cell supernatant. It significantly reduced malondialdehyde (MDA), increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and decreased intracellular ROS and nitric oxide (NO) content. Western blot analysis showed that the total flavonoids of Cymbidium fragrans decreased Bax levels in H9c2 cells damaged by H/R and increased Bcl-2 expression. Total flavones of Cymbidium fragrans upregulate VEGF-B/AMPK pathway related proteins VEGF-B, vascular endothelial growth factor receptor 1 (VEGFR-1), neuropilin 1 (NRP-1), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), phosphorylated adenosine monophosphate activated protein (p-AMPK) and phospho mechanistic target of rapamycin (p-MTOR) levels. The above research results indicate that the total flavonoids of Cymbidium can significantly reduce the H/R injury of myocardial cells, which may be related to the upregulation of VEGF-B/AMPK pathway and inhibition of oxidative stress response.

Analysis of a child with X-linked intellectual disability due to a maternal de novo splicing variant of the PAK3 gene / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic etiology for a child with profound intellectual disabilities and obvious behavioral abnormalities.@*METHODS@#A male child who had presented at the Zhongnan Hospital of Wuhan University on December 2, 2020 was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Short tandem repeat (STR) analysis was carried out to determine its parental origin. The splicing variant was also validated in vitro with a minigene assay.@*RESULTS@#WES results revealed that the child had harbored a novel splicing variant of c.176-2A>G in the PAK3 gene, which was inherited from his mother. The results of minigene assay have confirmed aberrant splicing of exon 2. According to the guidelines from the American College of Medical Genetics and Genomics, it was classified as a pathogenic variant (PVS1+PM2_Supporting+PP3).@*CONCLUSION@#The novel splicing variant c.176-2A>G of the PAK3 gene probably underlay the disorder in this child. Above finding has expanded the variation spectrum of the PAK3 gene and provided a basis for genetic counseling and prenatal diagnosis for this family.

Effect of salidroside on choroidal thickness and expression of hypoxia-inducible factor-1α, dopamine and its receptor in guinea pigs with myopia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To examine the effects of salidroside on choroidal thickness, hypoxia-inducible factor-1α(HIF-1α), dopamine(DA)and its D1 receptor expression in guinea pigs with lens-induced myopia(LIM).METHODS: A total of 18 two-week-old guinea pigs were randomly divided into the normal control(NC)group, the LIM group, and the LIM + salidroside(LIM+SA)group, with 6 guinea pigs in each group. The guinea pigs in the NC group were fed normally and intragastrically administered with 2 mL/d saline; those in the LIM group wore a -5D lens in front of their right eyes to establish a myopia model, then they were intragastrically administered with 2 mL/d saline. Finally, those in the LIM+SA group wore glasses along with intragastric administration of 2 mL/d salidroside at a dose of 100 mg/kg. The refraction, axial length, and choroidal thickness of guinea pigs in each group were measured 4wk following the establishment of the model. In addition, the relative mRNA expression and protein content of HIF-1α in the choroid and retina of guinea pigs in each group were detected by real-time quantitative PCR(qPCR)and immunohistochemistry(IHC). Finally, the DA concentration and its D1 receptor expression were detected by enzyme-linked immunosorbent assay(ELISA)and Western blot.RESULTS: At 4wk after model establishment, guinea pigs of LIM group and LIM+SA group exhibited increased negative refraction of the right eye, prolonged axial length, and decreased choroidal thickness compared to the NC group. The relative mRNA expression and protein content of HIF-1α in the choroid and retina of the guinea pigs increased. The concentration of DA and the expression of its D1 receptor both decreased. Moreover, compared to the LIM group, the diopter of the right eye of guinea pigs in LIM+SA group significantly reduced, the axial length was shorter, the thickness of choroid increased, the relative mRNA expression and protein content of HIF-1α in the choroid and retina decreased and the concentration of DA and the expression of its D1 receptor both increased.CONCLUSION: Salidroside can delay myopia progression in myopic guinea pigs by affecting choroidal thickness and the expression of HIF-1α, DA and its D1 receptor.

Research update on DNA methylation in peripheral blood cells as a risk factor for coronary heart disease / 中华检验医学杂志

Coronary heart disease (CHD) is a kind of cardiovascular diseases originated from atherosclerosis (AS), and chronic inflammation is one of the pathological characteristics. The peripheral blood leukocytes, especially mononuclear cells, play an important role in the AS processes. Recently, in a series of Epigenome-Wide Association Studies (EWAS), multiple DNA differential methylation sites in peripheral blood cells were found to be statistically associated with CHD, which suggested that these DNA differential methylation sites might serve as new risk factors for CHD. The recognition of the variant of DNA methylation as a common epigenetic nucleic acid modification in the occurrence and development of CHD, is ongoing. DNA methylation has the potential to become warning biomarkers, which might provide new ideas and evidences for mechanistic studies of CHD.

Anatomical and histological investigation of the area anterior to the anorectum passing through the levator hiatus / 中华胃肠外科杂志

Objective: To document the anatomical structure of the area anterior to the anorectum passing through the levator hiatus between the levator ani slings bilaterally. Methods: Three male hemipelvises were examined at the Laboratory of Clinical Applied Anatomy, Fujian Medical University. (1) The anatomical assessment was performed in three ways; namely, by abdominal followed by perineal dissection, by examining serial cross-sections, and by examining median sagittal sections. (2) The series was stained with hematoxylin and eosin to enable identification of nerves, vessels, and smooth and striated muscles. Results: (1) It was found that the rectourethralis muscle is closest to the deep transverse perineal muscle where the longitudinal muscle of the rectum extends into the posteroinferior area of the membranous urethra. The communicating branches of the neurovascular bundle (NVB) were identified at the posterior edge of the rectourethralis muscle on both sides. The rectum was found to be fixed to the membranous urethra through the rectourethral muscle, contributing to the anorectal angle of the anterior rectal wall. (2) Serial cross-sections from the anal to the oral side were examined. At the level of the external anal sphincter, the longitudinal muscle of the rectum was found to extend caudally and divide into two muscle bundles on the oral side of the external anal sphincter. One of these muscle bundles angled dorsally and caudally, forming the conjoined longitudinal muscle, which was found to insert into the intersphincteric space (between the internal and external anal sphincters). The other muscle bundle angled ventrally and caudally, filling the gap between the external anal sphincter and the bulbocavernosus muscle, forming the perineal body. At the level of the superficial transverse perineal muscle, this small muscle bundle headed laterally and intertwined with the longitudinal muscle in the region of the perineal body. At the level of the rectourethralis and deep transverse perineal muscle, the external urethral sphincter was found to occupy an almost completely circular space along the membranous part of the urethra. The dorsal part of the external urethral sphincter was found to be thin at the point of attachment of the rectourethralis muscle, the ventral part of the longitudinal muscle of the rectum. We identified a venous plexus from the NVB located close to the oral and ventral side of the deep transverse perineal muscle. Many vascular branches from the NVB were found to be penetrating the longitudinal muscle and the ventral part of rectourethralis muscle at the level of the apex of the prostate. The rectourethral muscle was wrapped ventrally around the membranous urethra and apex of the prostate. The boundary between the longitudinal muscle and prostate gradually became more distinct, being located at the anterior end of the transabdominal dissection plane. (3) Histological examination showed that the dorsal part of the external urethral sphincter (striated muscle) is thin adjacent to the striated muscle fibers from the deep transverse perineal muscle and the NVB dorsally and close by. The rectourethral muscle was found to fill the space created by the internal anal sphincter, deep transverse perineal muscle, and both levator ani muscles. Many tortuous vessels and tiny nerve fibers from the NVB were identified penetrating the muscle fibers of the deep transverse perineal and rectourethral muscles. The structure of the superficial transverse perineal muscle was typical of striated muscle. These findings were reconstructed three-dimensionally. Conclusions: In intersphincteric resection or abdominoperineal resection for very low rectal cancer, the anterior dissection plane behind Denonvilliers' fascia disappears at the level of the apex of the prostate. The prostate and both NVBs should be used as landmarks during transanal dissection of the non-surgical plane. The rectourethralis muscle should be divided near the rectum side unless tumor involvement is suspected. The superficial and deep transverse perineal muscles, as well as their supplied vessels and nerve fibers from the NVB. In addition, the cutting direction should be adjusted according to the anorectal angle to minimize urethral injury.

Age-related changes for the predictors of benign prostatic hyperplasia in Chinese men aged 40 years or older / 亚洲男科学杂志(英文版)

A cross-sectional study was conducted to estimate the age-stratified normal levels and age-related changes in the risk predictors of benign prostatic hyperplasia (BPH) progression. A total of 4706 male participants aged 40 years or older in Zhengzhou (China) were enrolled. The values of the International Prostate Symptom Score (IPSS), prostate-specific antigen (PSA), prostate volume (PV), and postvoid residual urine volume (PVR) significantly increased with age. Nonlinear relationships between age and IPSS scores ≥8 (P for nonlinearity = 0.046), PSA level ≥1.6 ng ml-1, PV ≥31 ml, or PVR ≥39 ml (all P for nonlinearity <0.001) were observed. After the age of 61 years, the risk indicators related to BPH progression were positively correlated with age (odds ratio [OR] >1), regardless of the predictors of the IPSS score, PSA level, PV, or PVR; and the OR values increased gradually. Therefore, after the age of 61 years, the risk predictors related to BPH progression were positively correlated with age.

Impact of the Varian real-time position management respiratory gating system on radiotherapy planning dosimetry / 中华放射医学与防护杂志

Objective:To study the impact of the Varian real-time position management (RPM) respiratory gating system on radiotherapy planning dosimetry.Methods:The radiotherapy plans of 40 cases with thoracic or abdominal tumors were retrospectively selected in this study. The motion phantom for quality control was adopted to generate respiratory gating signals, and the 30%-60% stable phase at the end of expiratory was selected as the respiratory gating window. The dose verification for the abovementioned radiotherapy plans was performed using the Portal Dosimetry (PD) system under RPM respiratory gating mode with the Edge accelerator. Afterwards, dose analysis was performed with different γ passing rate criteria and the distribution characteristics of γ values were analyzed. Finally, the verification results between the non-gating mode and the gating mode were compared.Results:Under the respiratory gating mode, the passing rates of all intensity-modulated radiation therapy/volumetric-modulated arc therapy (IMRT/VMAT) plans with or without flattening filters were over 95.5% by γ criteria of (3%, 3 mm) or (3%, 2 mm) and were over 90% by stricter γ criteria of (2%, 2 mm). All plans met the clinical requirements recommended by the American Association of Physicists in Medicine (AAPM). The passing rates of dose verification under non-gating mode were slightly better than those under respiratory gating mode, and the differences between the two modes were statistically significant (3%/3 mm, Z =-1.45; 3%/2 mm, Z =-2.86; 2%/2 mm, Z =-3.70; 1%/1 mm, Z =-4.52; P<0.05). There was no significant difference in the minimum and maximum values of γ and the share of γ > 1.5 of plan verification result under the two modes. However, the average value and standard deviation of the γ were generally smaller under the non-gating mode. Conclusions:The impact of the introduction of RPM respiratory gating technology on dose is clinically acceptable, and the execution of these plans in this gating mode is safe and reliable.

Effect of effective parts of Dracocephalum moldavica on the necroptosis of HBMECs after OGD/R injury / 药学学报

We investigated the ability of Dracocephalum moldavica (EPDM) flavonoids to protect human brain microvascular endothelial cells (HBMECs) from necroptosis induced by ischemia-reperfusion injury. To mimic the process of cerebral ischemia-reperfusion injury, a necroptosis model was established by treatment with the pan-cysteine aspartic acid protease (caspase) inhibitor Z-VAD-FMK combined with oxygen-glucose deprivation/re-oxygenation (OGD/R) injury using HBMECs. Cell proliferation and cytotoxicity (cell counting kit-8, CCK-8) was used to measure cell viability. A Hoechst33342/PI fluorescent double-staining method was exploited to determine the rate of cell necroptosis. A commercial kit was used to detect lactate dehydrogenase in the cell culture supernate. DCFH-DA probes, calcein AM and JC-1 probes were used to measure changes in ROS production, mitochondrial membrane permeability transformation pore (MPTP) opening and mitochondrial membrane potential (MMP), respectively. Enzyme-linked immunosorbent assay (ELISA) kits were chosen to detect the release of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6). Western blotting was used to detect necroptosis-related proteins. The results show that relative to control group, Z-VAD-FMK combined with OGD/R injury reduced cell viability, increased the necroptosis rate and the levels of LDH and ROS in HBMECs. The MPTP of the model group cells opened and the MMP reduced. TNF-α, IL-1β, and IL-6 levels were significantly elevated. Furthermore, the expression of receptor-interacting protein kinase 3 (RIP3) and mitochondrial phosphoglycerate mutase 5 (PGAM5) was significantly increased, accompanied by an increase of phosphorylated mixed-lineage kinase domain-like protein (p-MLKL)/MLKL. EPDM partially reversed the changes of the above-mentioned factors in HBMECs induced by Z-VAD-FMK plus OGD/R injury. These results indicate that EPDM may protect HBMECs from cerebral ischemia-reperfusion injury by inhibiting the RIP3/MLKL/PGAM5 pathway and MPTP opening to maintain mitochondrial function, thereby providing a scientific basis for the use of EPDM in the treatment of cerebral ischemia-related diseases.

Anatomical observation and clinical significance of the prostatic part of neurovascular bundle in total mesorectal excision / 中华胃肠外科杂志

Objective: To observe the anatomical architecture of the prostatic part of the neurovascular bundle (NVB) in total mesorectal excision (TME). Methods: A descriptive cohort study and an anatomical observation study were carried out. A total of 38 male patients with rectal cancer who underwent TME in the Department of Colorectal Surgery at the affiliated Union hospital of Fujian Medical University between November 2013 and March 2015 were included. A total of 4 hemipelvis were examined at the Laboratory of Clinical Applied Anatomy, Fujian Medical University. The following outcomes were observed: 1) the clinical significance of bleeding of the prostatic part of NVB: surgical videos were reviewed and the incidence of bleeding was recorded. The urogenital function was assessed using the International Prostate Symptom Score (IPSS) and International Index of Erectile Function (IIEF) score. The correlation between prostatic part bleeding and postoperative urogenital function was evaluated. 2) anatomical observation: the vessels, nerve fibers, as well as their surrounding fatty tissue from the prostatic part were treated as a whole, namely, the fat pad of the prostatic part. The anatomical architecture of the prostatic part in the surgical videos was reviewed and interpreted with the cadaveric findings. Categorical variables were compared between groups using a Fisher exact probability. while continuous variables with skewed distribution were compared between groups using the Mann-Whiteny U test. Results: The median age of the included 38 patients was 57 years (range, 31-75), and the median tumor distance to the anal verge was 6 cm (range, 1-8). Of them, a total number of 21 (55.3%) patients had bleeding of the prostatic part of NVB (bleeding group), while the rest had not (17 cases, 44.7%, non-bleeding group). 1) the clinical significance of bleeding of the prostatic part of NVB. The urinary function significantly decreased in patients in the bleeding group according to IPSS score after the 3rd month and the 6rd month of the surgery [7 (0-16) vs. 2 (0-3), Z=-1.787, P=0.088; 2 (0-15) vs. 0 (0-2), Z=-2.270, P=0.028]. There was no difference regarding the IPSS score between the two groups after 1 year of the surgery (P>0.05). With a total of 23 patients with normal preoperative sexual activity included, 87.5% (7/8) of patients in the non-bleeding group can expect to return to their preoperative baseline, this incidence was significantly higher than that of only 40% (6/15) in the bleeding group (P=0.029). 2) anatomical observation: for cadaveric observation, the prostatic part of NVB was located in the narrow triangular space composed of anterolateral walls of the rectum, the posterolateral surface of the prostate and the medial surface of the levator ani musculature. The tiny vascular branches and nerve fibers from the prostatic part were hard to identify. The cavernosal nerves cannot reliably be distinguished from the neural supply to the prostate, rectum and levator ani. In the cross-section of levels of prostatic base and mid-prostate in cadaveric hemipelvis specimens, the boundary of the prostatic part fat pad was partly overlapped and merged with the boundary of the mesorectum. Intraoperative observation showed that the areas of overlap referred to the rectal branches from the prostatic part piercing the proper fascia to supply the mesorectum, which carried the largest tension and high risk of bleeding during circumferential dissection toward the perirectal plane. The ultrasonic scalpel was required to pre-coagulate the rectal branches at the point close to the proper fascia of the rectum to prevent bleeding. In the cross-section of the prostatic apex level, the prostatic part approached ventrally and its boundary was away from the boundary of the mesorectum. Conclusions: NVB prostatic part injury is one of the causes of urogenital dysfunction after TME. The nerve fibers from the prostatic part were tiny, and its functional zones cannot be distinguished during operation. Therein, the fat pad of the prostatic part should be protected as a whole. Understanding the morphology of the fat pad of the prostatic part provides invaluable surgical guidance to dissect this critical area. When dissecting around the anterolateral rectal wall, appropriate anti-traction tension should be maintained and the rectal branches from the prostatic part should be coagulated with an ultrasonic scalpel to prevent bleeding.

Establishment of Drug-Resistant Cell Lines of Acute Myeloid Leukemia and Correlation of Sirt1 and PGC-1α Expression Levels with Drug Resistance / 中国实验血液学杂志

OBJECTIVE@#To construct cytarabine-resistant acute myeloid leukemia (AML) cell lines, and explore the correlation between Sirt1, PGC-1α expression levels and drug resistance.@*METHODS@#Human acute promyelocytic leukemia Kasumi-1 cells were induced by the method of gradually increasing the concentration of Ara-C drug. The IC50 value of Kasumi-1 cells before and after drug addition was detected by CCK-8 method, so as to construct Ara-C resistant cell lines. The expression levels of Sirt1 and PGC-1α mRNA in Kasumi-1 drug-resistant cell lines and their parental cell lines were detected by real-time fluorescence quantitative PCR, and the expression levels of Sirt1 and PGC-1α protein in kasumi-1 drug-resistant cell lines and their parental cell lines were detected by Western blot.@*RESULTS@#The constructed Kasumi-1 cell line had common morphological characteristics of drug-resistant cell lines under microscope, and the drug resistance index was greater than 5, indicating that Kasumi-1 drug-resistant cells had good drug resistance after the construction. The RT-qPCR and Western blot assays showed that the expression levels of Sirt1 and PGC-1α mRNA and protein in the drug-resistant cell lines were higher than those of the parental cell lines (P<0.001).@*CONCLUSION@#AML cell lines resistant to Ara-C can be successfully induced by the method of gradually increasing the concentration, and the co-high expression of Sirt1 and PGC-1α may mediate the drug resistance of AML cells.

Method of preparing macrophage-targeting nanobody 68Ga-NOTA-Nb119 as PET molecular probe / 西安交通大学学报(医学版)

【Objective】 To prepare positron emission computed tomography (PET) molecular probes targeting macrophages using Nb119 and BCⅡ10 labeled with 68Ga nuclide. 【Methods】 To explore the labeling conditions of nanobodies with 68Ga nuclides, first, the anti-Vsig4 nanobody Nb119 and isotype control antibody BCⅡ10 were incubated with NOTA and then purified to obtain the chelating agent-modified NOTA-Nb119 and NOTA-BCⅡ10. The NOTA-Nb119 and NOTA-BCⅡ10 were further incubated with 68Ga. Finally, NOTA-Nb119 was identified by SDS-PAGE and MALDI-TOF methods, and the radiochemical purity was detected by radio-HPLC. 【Results】 The results of SDS-PAGE showed that the lanes of the successfully labeled NOTA-Nb119 had apparent hysteresis than the unlabeled nanobody band, which proved that the molecular weight of the labeled product was increased and NOTA successfully modified the nanobody. Subsequently, 68Ga nuclides could successfully label Nb119 and BCⅡ10. According to radio-HPLC detection, the radiochemical purity of NOTA-Nb119 and NOTA-BCⅡ10 labeled with 68Ga was greater than 90% and remained stable. 【Conclusion】 68Ga-labeled nanobodies have high radiochemical purity and high stability, indicating that this study can be applied to other nanobodies for modification and labeling and provides a practical and feasible method for the preparation of PET using nanobodies.

Study of the protective effect of total flavonoids of Dracocephalum moldavica L.On doxorubicin induced injury in HUVECs based on VEGF-B/AMPKa pathway / 中国药理学通报

Aim To investigate the protective effect of TFDM on doxorubicin-induced endothelial cell injury and its mechanism.Methods Cell viability was detected by CCK-8 assay.Cell morphology was observed by microscope.The changes of LDH, SOD and mitochondrial membrane potential were detected by kit method.Cell migration was detected by Transwell assay; Endothelial dysfunction and VEGF-B/AMPKa pathway related protein expression were detected by Western blot.Results Compared with model group, TFDM significantly increased cell viability, improved the morphologic changes of HUVEC induced by DOX, decreased LDH leakage, increased SOD activity, increased mitochondrial membrane potential, promoted endothelial cell migration, and inhibited endothelial cell injury.The results of Western blot showed that com pared with control group TFDM increased the expression levels of non-receptor tyrosine kinase ( Src) and focal adhesion kinase (FAK) .increased the phosphorylation level of eNOS, and decreased the expression level of ET-1 protein, thereby inhibiting endothelial dysfunction.The protein expression levels of VEGF-B, NRP1 , VEGFR1 and the ratio of p-AMPKa/AMPKa significantly increased in the administration group.Conclusion TFDM may inhibit doxorubicin-induced endothelial cell injury by activating VEGF-B/AMPKa pathway.

Basic and clinical treatments target tumor-associated macrophages in liver cancer / 西安交通大学学报(医学版)

Hepatocellular carcinoma （HCC） most often develops in patients with liver disease characterized by chronic non-resolving inflammation. The inflammatory response is mainly derived from innate immune cells， and tumor-associated macrophages （TAMs） play an important role in the development of tumors．It is usually chemotactic from mononuclear progenitor cells in the blood to tumor tissue， then is induced by the tumor microenvironment and further develops into TAMs. They play an important role in promoting tumor growth，angiogenesis and tumor invasion and inducing tumor tissues to form an environment without inhibition mechanisms. As the relationship between TAMs and malignant tumors becomes clearer， TAMs are beginning to be seen as therapeutic targets. The heterogeneity of TAM subtypes and their origin and dynamic phenotype during the initiation and progression of HCC has been partially unraveled. It further forms the base for developing therapeutic agents by decreasing the population of TAMs via blocking recruitment of bone marrow-derived monocytes and functionally reprograms TAMs to anti-tumoral behavior. This review focuses on the preclinical evolution and hitherto clinical trials for TAM-targeted therapy in HCC.

Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells / 中草药·英文版

Objective: Chrysophanol (Chry) displays potent anticancer activity in human cancer cells and animal models, but the cellular targets of Chry have not been fully defined. Herein, we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity. Methods: Human liver cancer cell line HepG2 was incubated. The cytotoxicity was evaluated by MTT assay. Mitochondria localization was evaluated by a confocal microscopy. Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer. The levels of ATP, mitochondrial superoxide anions, and GSH/GSSG were determined according to the assay kits. The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining, respectively. The expression of cyclophilin D (CyPD) was determined by immunoblot method, and the interaction between CyPD and Chry was analyzed by molecule docking procedure. Results: Chry itself mainly localized in mitochondria to cause mitochondrial dysfunction and cell death in HepG2 cells. As regard to the mechanism, cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore (mPTP) moderately suppressed cell death, indicating mPTP involved in the process of cell death. Further, Chry enhanced the protein expression of Cyclophilin D (CyPD) which is a molecular componentry and a modulator of mPTP, while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD. Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with −11.94 kal/mol of the binding affinity value. Besides, the mtDNA-deficient HepG

Ethanol extract of Ardisiae Japonicae Herba inhibits hepatoma carcinoma cell proliferation in vitro through regulating lipid metabolism / 中草药·英文版

Objective: The aim of this study is to discover the possible working mechanisms of Ardisiae Japonicae Herba (AJH) on hepatoma carcinoma (HCC). Methods: In this study, ethanol extract of AJH was prepared and used to treat HCC cell in vitro. Furthermore, a genomic wide RNA sequencing (RNA-seq) was performed to screen deregulated genes in HCC cells after the treatment of AJH extract. The gene and protein expression related to lipid metabolism in HCC cells were also investigated to validate the results obtained from RNA-seq. Results: AJH extract could inhibit HCC cell proliferation in vitro. RNA-seq analysis has identified 1,601 differentially expressed genes (DEGs, fold change ≥ 2.0 or fold change ≤ 0.5, P < 0.05) in HCC after AJH extract treatment, which included 225 up-regulated genes and 1,376 down-regulated genes. KEGG pathway analysis of DEGs demonstrated that lipid metabolism was a potential pathway related to AJH treatment. In agreement with the RNA-seq data, qPCR and Western-blot analysis indicated that expression of genes and proteins related to lipid metabolism (SREBP1, ACC, ACLY and FASN) were significantly down-regulated in AJH treatment group as compared with the control group. Furthermore, AJH extract could also decrease lipid contents and cellular free fatty acid levels in HCC cells. Conclusion: Ethanol extract of AJH could inhibit HCC cell proliferation in vitro, the possible mechanism may be related to the inhibition of lipid metabolism.

Anatomical observation of the right retroperitoneal fascia and its clinical significance in complete mesocolic excision for right colon cancer / 中华胃肠外科杂志

Objective: To investigate the anatomic characteristics of the right retroperitoneal fascia and its surgical implementation while performing complete mesocolic excision (CME) for right colon cancer. Methods: A descriptive study was carried out. (1) Clinicopathological data and surgical videos of 17 non-consecutive patients undergoing laparoscopic right hemicolectomy (extended right hemicolectomy) with CME for right colon cancer at Department of Colorectal Surgery of Union Hospital, Fujian Medical University between January 2020 and October 2020 were retrospectively collected. The construction of right retroperitoneal fascia was observed from caudal dorsal direction and caudal ventral direction. (2) Three postoperative specimens from 3 cases undergoing laparoscopic right hemicolectomy with CME for right colon cancer in June 2020 were prospectively included to observe anatomy and examine histology. (3) Five abdominal cadaver specimens from the Department of Anatomy of Fujian Medical University were enrolled, including 3 males and 2 females. Anatomical observation and histological studies were performed from the cranial approach and the caudal dorsal approach. Masson staining was used to examine the histology. Results: (1) Surgical video observation: The typical structure of right retroperitoneal fascia could be observed in all the 17 patients. The fascia was a rigid barrier between the posterior space of the ascending colon and the anterior pancreaticoduodenal space behind the transverse colon. The right retroperitoneal fascia should be sharply cut to communicate between the two spaces to avoid entering the right mesocolon by mistake. The severed ventral stump of the right retroperitoneal fascia ran along the dorsal side of the right hemicolon to the lateral side, and the dorsal stump covered the level of the duodenum caudally, and continued to move downward, covering the surface of Gerota's fascia. (2) Observation of 3 surgical specimens: The dorsal side of the right mesocolon was smooth and intact, which could be anchored in the corresponding area of the lateral edge of the duodenum. The ventral stump of the right retroperitoneal fascia could be seen, which attached to the dorsal side of the right mesocolon semi-circularly. Masson staining observation: The ventral stump of the right retroperitoneal fascia ran cephalad, fused with the dorsal side of the right mesocolon tightly and curled. The caudal side of confluence and the dorsal side of the right mesocolon presented a bilobed structure. (3) Anatomy of 5 cadaveric specimens: The right retroperitoneal fascia was a thin fascia structure, which was a rigid barrier between the anterior pancreaticoduodenal space behind the transverse colon and the posterior space of the ascending colon. The ventral stump of the right retroperitoneal fascia (including the dorsal side of the right mesocolon), the dorsal stump of the right retroperitoneal fascia (including part of the duodenal wall) and the dorsal side of the right mesocolon were retrieved for histological examination. The ventral stump of the right retroperitoneal fascia fused with the dorsal side of the right mesocolon by the cephalic side, and the dorsal side of the right hemi-mesocolon on the fusion level by caudal side gradually separated into a double-layer loose fascial structure. The dorsal stump of the right retroperitoneal fascia covered the surface of the duodenum level, moved on from the ventral side to the surface of the prerenal fascia, and continued to the caudal side. Conclusions: The right retroperitoneal fascia is a rigid barrier between the anterior pancreaticoduodenal space behind the transverse colon and the posterior space of the ascending colon. The Toldt fascia formed by fusion with the dorsal lobe of the right colon travels to the edge of the descending and horizontal part of the duodenum and separates again. The right retroperitoneal fascia is attached to the edge of the duodenum, reversing and running on the surface of the prerenal fascia, while the dorsal lobe of the right colon runs in front of the pancreas and duodenum, and shifts to the pancreaticoduodenal fascia. During the operation, this fascia should be identified and cut to penetrate the anterior pancreaticoduodenal space behind the transverse colon and the posterior ascending colon space, which helps to ensure the integrity of the dorsal side of the right hemi-mesocolon.

Anatomical observation of the left parietal peritoneum and its clinical significance in left retro-mesocolic space dissection / 中华胃肠外科杂志

Objective: To investigate the anatomic characteristics of the left parietal peritoneum and its surgical implementation while dissecting in left retro-mesocolic space. Methods: A descriptive case series research methods was used. (1) surgical videos of 35 patients who underwent laparoscopic radical resection (complete mobilization of splenic flexure) of colorectal cancer in Union Hospital of Fujian Medical University between January 2018 and December 2018 were reviewed; (2) four specimens after radical resection of rectal cancer performing in June 2020 were prospectively enrolled and reviewed; (3) five specimens of left parietal peritoneum from 5 cadaveric abdomen (3 males and 2 females) were enrolled and reviewed as well; Tissues of 3 unseparated regions, namely the root of the inferior mesenteric artery (IMA), the medial region and the lateral region (including kidney tissue), from above the 5 cadaveric abdominal specimens were selected to perform Masson staining and histopathological examination. Results: (1) Surgical video observation: "Staggered layer phenomenon" and typical left parietal peritoneum was found in 77.1% (27/35) of patients when the left retro-mesocolic space was separated from the lateral and central approaches. The left parietal peritoneum presented as a rigid fascia barrier between the lateral and central approaches, which was a translucent dense connective tissue fascia. After the splenic flexure were completely mobilized, the left parietal peritoneum stump continued to the cephalic side. (2) Observation of 4 surgical specimens: The dorsal side of the left mesocolon specimen was studied, and the left parietal peritoneum stump edge was identified. The outside of the stump edge was the left hemicolon dorsal layer, which was continuously downward to the rectal fascia propria. (3) Cadaveric abdominal specimens: The left retro-mesocolic space was separated through lateral and central approaches, and the rigid fascia barrier, essentially the left parietal peritoneum and Gerota fascia, was encountered. Cross-section view showed that the left parietal peritoneum could be further detached from the dorsal layer of the left mesocolon from the outside, but could not be further detached from the inside out. (4) Histological examination: There was no obvious fascia structure in the IMA root region, while outside the IMA root region, the left bundle of inferior mesenteric plexus penetrating Gerota fascia was observed. There were 4 layers of fascias in the medial region, including the ventral layer of the left mesocolon, the dorsal layer of the left mesocolon, left parietal peritoneum and Gerota fascia. Small vessels were observed between the dorsal layer of the left mesocolon and the left parietal peritoneum. In lateral region, renal tissue and renal fascia were observed. Three layers of fascia structures were observed clearly under high power field, including the dorsal layer of the left mesocolon, left parietal peritoneum, and Gerota fascia. Conclusions: The left parietal peritoneum is the anatomical basis of the "staggered layer phenomenon" from the lateral or central approaches during the separation of left retro-mesocolic space. The small vessels in the dissection plane are the anatomical basis of intraoperative microbleeding, which need pre-coagulation. The central part of Gerota fascia is penetrated by the branches of the inferior mesenteric plexus, which results in a relatively dense surgical plane. Thus, during the dissection through the central approach, it is easy to involve in wrong surgical plane by deeper dissection.

Epidemiological characteristics and disease spectrum analysis of outpatients in the dermatology department of a hospital in Wuhan during the COVID-19 epidemic / 中华皮肤科杂志

Objective:To investigate epidemiological characteristics of outpatients and disease spectrum in the dermatology department during the COVID-19 epidemicMethods:A retrospective comparison of outpatient visits, gender, age and disease types in the dermatology department of Wuhan No.1 Hospital was performed between COVID-19 epidemic period (from 23th January 2020 to 15th April 2020) and the same period in 2019. Enumeration data were analyzed by Pearson′s chi-square test.Results:During the COVID-19 epidemic, the number of outpatient visits to the dermatology department of the hospital decreased markedly, and the average daily number of outpatient visits (236 visits/day) was only 8.81% of that during the same period in 2019 (2 678 visits/day) ; the ratio of male to female patients was reversed from 1∶1.37 in 2019 to 1.16∶1 in 2020; the proportions of patients aged 0-6, 7-12, 13-17 and 18-45 years significantly decreased compared with those in 2019 (all P < 0.001) , and the proportions of patients aged 46-69 and > 69 years significantly increased (both P < 0.001) . During the COVID-19 epidemic, there were 171 types of skin diseases in the dermatology outpatient department, and the number of disease categories decreased compared with that during the same period in 2019 (442 types) ; the number of patient visits for allergic skin diseases, erythematous papulosquamous skin diseases, viral infectious skin diseases and bacterial infectious skin diseases significantly increased compared with that during the same period in 2019 (all P < 0.001) , while the number of patient visits for sebaceous and sweat gland disorders, pigmented skin diseases and physical skin diseases significantly decreased (all P < 0.001) . Conclusion:Compared with the same period in 2019, the number of outpatient visits, patient sex ratio, age distribution and disease types in the dermatology department have undergone marked changes during the COVID-19 epidemic, and this study provides a reference for healthcare workers in dermatology department to respond to various epidemics and natural disasters in the future.

Study on serum tumor markers in retinoblastoma / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the secretion levels of serum tumor markers neuron specific enolase(NSE), glycoprotein antigen CA153 and CA199 in patients with retinoblastoma(RB).<p>METHODS: Data of 42 RB patients who received chemotherapy at Shenzhen People's Hospital between October 2017 and October 2019 were collected. The RB group was regrouped according to early and mid-term/advanced stages, and single/double eye involvement, the levels of serum tumor markers were compared between these subgroups and different genders.<p>RESULTS: The levels of tumor markers NSE, CA153 and CA199 in the advanced group were higher than those in the early and mid-term groups(49.69±18.45ng/mL <i>vs</i> 36.18±14.92ng/mL, 22.38±12.03U/mL <i>vs</i> 15.10±8.32U/mL, 46.44±18.76U/mL <i>vs </i>30.21±24.03U/mL, <i>P</i><0.05), but there was no statistically significant difference in expression between the groups of different genders and single and binocular involvement(<i>P</i>>0.05).<p>CONCLUSION: Serum NSE, CA153 and CA199 are significantly increased in advanced group than early and mid-term group, and they are of great significance in the diagnosis and treatment of RB. The difference between NSE in single and double eyes is worthy of further study.