BACKGROUND:The specific mechanism of Gushukang,as a Chinese traditional patent medicine for the treatment of postmenopausal osteoporosis of kidney deficiency and blood stasis,needs further studies. OBJECTIVE:To investigate the effect of Gushukang on serum sex hormones,bone microstructure and estrogen receptor in postmenopausal osteoporosis. METHODS:Firstly,network pharmacological analysis was performed.The active ingredients and action targets of Gushukang and the targets of postmenopausal osteoporosis were obtained respectively.Cytoscape was used to construct the active ingredient-target network.STRING database and Cytoscape were used for protein-protein interaction analysis and screening of core targets.DAVID database was used for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of intersection targets.Then the ovariectomized Sprague-Dawley rats were used in the animal experiment.Gushukang was administered by gavage for 3 months.The serum estrogen level was detected by ELISA,the bone microstructure was detected by microCT,and the protein expression of estrogen receptor α and estrogen receptor β in bone tiusse was detected by western blot. RESULTS AND CONCLUSION:The network pharmacological research results identified 132 active ingredients and 150 targets of Gushukang and 1155 targets of postmenopausal osteoporosis.After intersections with 1155 postmenopausal osteoporosis targets,87 targets of active ingredients of Gushukang against postmenopausal osteoporosis were obtained.By constructing the active ingredient-target network,it was found that the active ingredients at the core were quercetin,kaempferol,luteolin,naringin and isorhamnetin,and the targets at the core were NCOA2,ESR2,AR,F2,ESR1 and PTGS1.The final targets obtained after the protein-protein interaction analysis and screening included MAPK8,ESR1,JUN,R3C1,RELA and FOS,of which ESR1 was the common core target obtained from the two analyses.KEGG enrichment analysis showed estrogen,tumor necrosis factor,apoptosis and other signaling pathways.Therefore,animal experiments focused on the effect of Gushukang on different subtypes of estrogen receptors in the estrogen signaling pathway.The results showed that in the Gushukang group,bone microstructure was significantly improved,serum estrogen level had no significant change,but the protein expression of estrogen receptor α and β in bone tissue was significantly increased.All the findings indicate that the mechanism of Gushukang in the treatment of postmenopausal osteoporosis may be related to its hormone-like effect and the enhancement of estrogen receptor expression.

BACKGROUND:Current studies have confirmed that Ganoderma lucidum polysaccharides can promote nerve regeneration in neurodegeneration-related diseases.The occurrence of neurodegenerative diseases is closely related to mitochondrial dysfunction,but the role of Ganoderma lucidum polysaccharides on the regulation of apoptosis and mitochondrial function in neurodegenerative diseases is not yet clarified. OBJECTIVE:To explore the regulatory effects and mechanisms of Ganoderma lucidum polysaccharides on apoptosis and mitochondrial dysfunction in H2O2-induced SH-SY5Y cells. METHODS:SH-SY5Y cells were divided into three groups:control group,H2O2 group,and Ganoderma lucidum polysaccharides group.Cells in the control group were normally cultured.Cells in the H2O2 group were treated with 300 μmol/L H2O2 for 24 hours.In the Ganoderma lucidum polysaccharides group,the intervention with 300 μg/L Ganoderma lucidum polysaccharides was conducted first for 1-2 hours,followed by the addition of 300 μmol/L H2O2 for 24 hours.The mitochondrial membrane potential was detected by JC-1 kit.Apoptosis was detected by TUNEL staining kit.The activities of malondialdehyde and superoxide dismutase were detected by malondialdehyde test kit and superoxide dismutase test kit,respectively.The apoptosis and expression of mitochondrial dynamics-related proteins were detected by immunofluorescence staining and western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the mitochondrial membrane potential and superoxide dismutase activity were significantly reduced,as well as apoptotic rate and malondialdehyde levels were significantly increased in the H2O2 group(P<0.05).After treatment with Ganoderma lucidum polysaccharides,the membrane potential and superoxide dismutase activities were significantly increased,and apoptotic rate and malondialdehyde levels were significantly reduced compared with the H2O2 group(P<0.05).(2)The expression levels of pro-apoptotic proteins Bax and Caspase-3 were significantly increased,but the expression of anti-apoptotic protein Bcl-2 was significantly decreased in the H2O2 group compared with the control group(P<0.05).Compared with the H2O2 group,the levels of Bax and Caspase-3 were significantly decreased,but the expression of anti-apoptotic protein Bcl-2 was significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(3)Compared with the control group,the expression of mitochondrial splitting proteins Fis1 and p-Drp1 was significantly increased,but the expression of mitochondrial fusion proteins OPA1,Mfn1,and Mfn2 was decreased in the H2O2 group(P<0.05).Compared with the H2O2 group,Fis1 and p-Drp1 expression was significantly reduced,but the expression levels of OPA1,Mfn1,and Mfn2 were significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(4)The above results confirm that Ganoderma lucidum polysaccharides can attenuate H2O2-induced oxidative stress damage and apoptosis in SH-SY5Y cells by ameliorating mitochondrial dysfunction.

ObjectiveTo observe the effect of Gualou Xiebai Banxiatang on mitochondrial dysfunction and adenosine 5'-monophosphate （AMP）-activated protein kinase （AMPK）/peroxlsome proliferator-activated receptor-γ coactlvator-1α （PGC-1α） signaling pathway in rats with ischemic myocardial injury. MethodSeventy male SD rats were used in this experiment. Six rats from them were randomly selected as the control （CON） group， and the others were given high fat diet combined with isoproterenol injection （5 mg·kg^-1·d^-1， 7 d） to induce the rat model of ischemic heart disease based on hyperlipidemia. Successfully modeled rats were then randomly divided into model （MOD） group， high-dose Gualou Xiebai Banxiatang （GXBD-H） group， medium-dose Gualou Xiebai Banxiatang （GXBD-M） group， low-dose Gualou Xiebai Banxiatang （GXBD-L） group， and metoprolol （MET） group. Rats in the GXBD-H， GXBD-M， and GXBD-L groups were given 11.2， 5.6， 2.8 g·kg^-1·d^-1 Gualou Xiebai Banxiatang， those in the MET group were given 2.6 mg·kg^-1·d^-1 metoprolol， and those in the CON and MOD groups were given equal volume of pure water for 28 d. Hemodynamics were measured in rats by cardiac catheterization. Transmission electron microscopy was used to analyze myocardial mitochondria. Serum brain natriuretic peptide （BNP） and cardiac troponin T （cTnT） levels were detected by enzyme-linked immunosorbent assay （ELISA）. Mitochondrial membrane potential assay kit （JC-1 method） was applied for detecting mitochondrial membrane potential. The changes in the mitochondrial DNA copy number were measured by real-time quantitative polymerase chain reaction （Real-time PCR）. The content of adenosine triphosphate （ATP） in myocardial tissues was determined by spectrophotometer. The expression levels of p-AMPK， AMPK， PGC-1α， nuclear respiratory factor 1 （NRF1）， and mitochondrial transcription factor A （TFAM） in myocardium was detected by Western blot. ResultAs compared with the CON group， left ventricular end-systolic pressure （LVESP） and left ventricular end-diastole pressure （LVEDP） in the MOD group were significantly increased （P<0.05， P<0.01）， and +dp/dt_max and -dp/dt_max were significantly decreased （P<0.01）. In the MOD group， cardiac index and myocardial interstitial fibrosis area were significantly increased （P<0.01）， accompanied by mitochondrial damage， serum BNP， cTnT， and malondialdehyde （MDA） were significantly increased （P<0.01）， and serum superoxide dismutase （SOD） level was significantly decreased （P<0.01）. The myocardial mitochondrial membrane potential， DNA copy number， and ATP level were significantly decreased （P<0.01）， and the protein expression levels of p-AMPK/AMPK， PGC-1α， NRF1， and TFAM in myocardial tissues were significantly decreased in the MOD group （P<0.01）. Compared with the MOD group， the GXBD-H and GXBD-M groups significantly improved LVESP， LVEDP， +dp/dt_max， and -dp/dt_max （P<0.05， P<0.01）， significantly decreased heart index and myocardial interstitial fibrosis area （P<0.05， P<0.01）， and alleviated mitochondrial damage. In the GXBD-H and GXBD-M groups， serum BNP， cTnT， and MDA were decreased significantly （P<0.05， P<0.01）， serum SOD level was increased significantly （P<0.05）， and myocardial mitochondrial membrane potential， DNA copy number， and ATP level were significantly increased （P<0.05， P<0.01）. The protein levels of p-AMPK/AMPK， PGC-1α， NRF1， and TFAM in myocardial tissues were significantly increased in the GXBD-H and GXBD-M groups （P<0.05， P<0.01）. ConclusionGualou Xiebai Banxiatang has the effects of reducing the changes in cardiac function and myocardial pathology of rats with myocardial injury， inhibiting mitochondrial dysfunction， and up-regulating the protein expression levels of p-AMPK/AMPK， PGC-1α， NRF1， and TFAM in myocardial tissues. This study provides new laboratory evidence for in-depth exploration of the mechanism of this classical compound in preventing and treating myocardial injury.

【Objective】 To investigate the effect of total flavone of oldenlandia diffusa(FOD) on the stemness, proliferation and apoptosis of breast cancer(BC) stem cells sorted from MDA-MB-231. 【Methods】 Human BC cell lines MDA-MB-231 was cultured in vitro; MDA-MB-231 was stimulated by different concentrations(0 μg/mL, 100 μg/mL, 200 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h and 72 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on MDA-MB-231 proliferation; CD44⁺/CD24^-MDA-MB-231 cell line were tested by flow cytometry and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting; Annexin V-PE/7-AAD was used to detect the effect of FOD on MDA-MB-231 apoptosis and Bcl2, cleaved-caspase3 and Bax were tested by Western blotting. 【Results】 Cell proliferation of MDA-MB-231 was significantly inhibited by FOD, with the significant suppression at concentrations of 400 μg/mL for 72 h compared with negative control group(P<0.05). The apoptosis rate was significantly upregulated than the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and cleaved-caspae3 increased, and stemness markers such as Nanog, Sox2 and Oct4 decreased in FOD-treated cells. Moverover, Akt-GSK3β-β-catenin axis was inhibited in FOD-treated cells. 【Conclusion】 FOD could significantly inhibit the stemness and proliferation and promote the apoptosis of MDA-MB-231.

【Objective】 To investigate the effects of total flavone of oldenlandia diffusa (FOD) on the proliferation and apoptosis of hepatocellular carcinoma (HCC) stem cells sorted from Huh7. 【Methods】 Human HCC cell lines Huh7 was cultured in vitro; CD133 positive (CD133⁺) stem cells in Huh7 cell line were sorted by flow cytometry, and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting. CD133⁺-Huh7 was stimulated by different concentrations (0 μg/mL, 50 μg/mL, 100 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h, 72 h and 96 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on CD133⁺-Huh7 proliferation while Annexin V-PE/7-AAD was used to detect the effect of FOD on CD133⁺-Huh7 apoptosis. Western blotting was used to detect the protein expressions of protein 53 (P53), factor associated suicide-Fas-associating protein with a novel death domain (Fas-FADD), B-cell lymphoma-2 (Bcl-2), Cleaved-Caspase3, and Bcl-2 associated X protein (Bax). 【Results】 More than 95% of stem cells were purified for further experiments. Cell proliferation of CD133⁺-Huh7 was significantly inhibited by FOD, with the significant suppression at the concentration of 100 μg/mL for 72 h compared with negative control group (P<0.05). The apoptosis rate was significantly upregulated than that in the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and Cleaved-Caspae3 increased via FAS/FADDD and P53 axis. 【Conclusion】 FOD can significantly inhibit the proliferation and promote the apoptosis of CD133⁺-Huh7.

Objective:To investigate the preventive effect of postoperative pancreatic fistula by pedicled omentum wrapping pancreaticojejunostomy in pancreaticoduodenectomy.Methods:The clinical data of 78 patients who underwent pancreaticoduodenectomy in Songjiang Hospital Affiliated to Shanghai Jiao Tong University from September 2017 to August 2020 were retrospectively analyzed. Patients were divided into the modified pancreaticojejunostomy group (conventional approach group, n=43) and the pedicled omentum wrapping pancreaticojejunostomy group (omentum wrapping group, n=35) according to the surgical mode. Postoperative pancreatic fistula, postoperative abdominal bleeding, delayed gastric emptying and postoperative hospital stay were compared between the two groups. Results:Compared with the conventional approach group, the postoperative pancreatic fistula rate in omentum wrapping group was significantly reduced (2.9% vs 18.6%), and the difference was statistically different ( P=0.030); postoperative abdominal bleeding rate(0 vs 4.7%)and the delayed gastric emptying rate were significantly reduced (8.6% vs 25.6%), but both of them were not statistically different ( all P value>0.05); the length of postoperative hospital stay was significantly reduced [(11.3±2.9) days vs (12.8±3.5)days], and the difference was statistically different ( P=0.045). Conclusions:The pedicled omentum wrapping pancreaticojejunostomy was simple and convenient, which can significantly reduce the rate of postoperative pancreatic fistula. It can shorten the time of the length of hospital stay after surgery.

Objective:To systematic review the efficacy and safety of external application of traditional Chinese medicine on treatment of breast cancer related lymphedema.Methods:CNKI, Wanfang Data, VIP, Sino-Med, The Cochrane Library, PubMed, Web of Science were searched for related randomized controlled trials, the retrieval time was from inception to May 25, 2020. Two researchers independently screened and evaluated the included studies. Meta-analysis was performed by RevMan 5.4 software.Results:A total of 16 studies involving 1 315 patients with breast cancer related lymphedema were included, and the methodological quality of the included studies was not high. Compared with conventional treatment, external application of traditional Chinese medicine combined with conventional treatment had advantages in improving the total efficiency( P<0.01) and quality of life( P<0.01), reducing pain( P<0.01) and improving upper limb function( P<0.01), without obvious adverse reactions( P>0.05), but there was no improvement in depression( P>0.05). Compared with conventional treatment, external application of traditional Chinese medicine could improve the total efficiency( P<0.01). Compared with placebo sticker combined with conventional treatment, external application of traditional Chinese medicine combined with conventional treatment can reduce circumference( P<0.05) and reduce pain( P<0.01), without obvious adverse reactions( P>0.05). Conclusions:Available evidence suggests that external application of traditional Chinese medicine may be a potential treatment method for breast cancer related lymphedema. Due to the poor methodological quality of the included studies, high-quality randomized controlled trials are still needed.

ObjectiveTo investigate the value of aspartate aminotransferase-to-platelet ratio index (APRI), fibrosis-4 (FIB-4) score, and gamma-glutamyl transpeptidase-to-platelet ratio (GPR) in diagnosis of liver inflammation grade in patients with chronic hepatitis B (CHB). MethodsA total of 545 patients with CHB who underwent percutaneous liver biopsy and routine laboratory examinations during hospitalization in Shanghai Public Health Clinical Center Affiliated to Fudan University from October 2016 to October 2019 were enrolled. Inflammation grade (G) was determined according to the Scheuer scoring system, and APRI, FIB-4, and GPR were calculated based on related clinical indicators. The t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. A Spearman correlation analysis was used to investigate the correlation between two variables. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic performance of the three serum noninvasive diagnostic models in determining liver inflammation grade, and the Delong test was used for comparison of the area under the ROC curve (AUC). ResultsAmong the 545 patients, 224 had grade G0-1 liver inflammation, 209 had grade G2 liver inflammation, and 112 had grade G3 liver inflammation. The Spearman correlation analysis showed that APRI, FIB-4, and GPR were positively correlated with liver inflammation grade (r=0.611, 0.470, and 0.563, all P＜0.001). APRI, FIB-4, and GPR had an AUC of 0.820, 0.719, and 0782, respectively, in the diagnosis of G≥2 liver inflammation, with optimal cut-off values of 0.53, 1.48, and 0.20, respectively; for the diagnosis of G≥2 liver inflammation, GPR had a better performance than FIB-4 (P=0.01) and a slightly lower performance than APRI (P=0.048). The stratified analysis based on alanine aminotransferase (ALT) level showed that in the ≤1×upper limit of normal (ULN) group, the (1-2)×ULN group, and the (2-5)×ULN group, APRI had an AUC of 0.847, 0.786, and 0.724, respectively, in the diagnosis of G≥2 liver inflammation, FIB-4 had an AUC of 0.777, 0.729, and 0.626, respectively, and GPR had an AUC of 0.801, 0.781, and 0.607, respectively; the subgroup analysis showed that GPR had a similar diagnostic performance to APRI and FIB-4 in all ALT stratification groups except the (2-5)×ULN group, in which GPR had a lower diagnostic performance than APRI (P=0.042). APRI, FIB-4, and GPR had an AUC of 0.791, 0.725, and 0.801, respectively, in the diagnosis of G≥3 liver inflammation, with optimal cut-off values of 0.66, 1.49, and 0.25, respectively; in the diagnosis of G≥3 liver inflammation, GPR had a similar diagnostic performance to APRI and a better diagnostic performance than FIB-4 (P=0.006). The stratified analysis based on ALT level showed that in the ≤1×ULN group, the (1-2)×ULN group, and the (2-5)×ULN group, APRI had an AUC of 0.900, 0.742, and 0.693, respectively, in the diagnosis of G≥3 liver inflammation, FIB-4 had an AUC of 0.874, 0.683, and 0.644, respectively, and GPR had an AUC of 0.890, 0.805, and 0.668, respectively. The subgroup analysis showed that GPR had a similar diagnostic performance to APRI and FIB-4 in all ALT stratification groups except the (1-2)×ULN group, in which GPR had a better diagnostic performance than FIB-4(P=0.015). ConclusionAPRI, FIB-4, and GPR may accurately diagnose liver inflammation grade in CHB patients, which helps to monitor the progression of CHB and determine the timing of antiviral therapy.

[摘 要] 目的：探讨肝内胆管癌（intrahepatic cholangiocarcinoma，ICCA）围手术期外周血中性粒细胞与淋巴细胞比率（neutrophil-to-lymphocyte ratio，NLR）和血小板与淋巴细胞比率（platelet-to-lymphocyte ratio，PLR）对患者预后的预测价值。方法：收集2015年1月至2018年1月在上海市松江区中心医院接受肝切除术治疗的ICCA患者97例作为ICCA组，选择同期在本院做健康体检的志愿者100例作为正常对照组。检测两组受试者术前1 d、术后3 d和7 d外周血的NLR、PLR，采用单因素、多因素分析ICCA患者术后随访期死亡的危险因素，采用Kaplan-Meier生存曲线分析术后3 d的NLR和PLR对ICCA患者术后生存时间的影响，采用受试者工作特征曲线（receiver operating characteristic curve，ROC）分析术后3 d的NLR和PLR水平对患者术后随访期死亡的预测价值。结果：ICCA组患者术前1 d、术后3和7 d外周血的NLR、PLR均高于正常对照组（均P<0.05），术前1 d和7 d外周血NLR、PLR差异无统计学意义（P>0.05），术后3 d外周血NLR、PLR水平最高（P<0.05）。多发肿瘤、合并淋巴结转移、TNM分期Ⅲ~Ⅳ、CA199水平增高、术后3 d的NLR和PLR较高分别是ICCA患者随访期死亡的独立危险因素（均P<0.05）。ROC曲线显示，术后3 d的NLR和PLR高低对ICCA患者术后生存时间具有预测价值。Kaplan-Meier生存曲线显示，低NLR[（50.32±3.69） vs （30.12±2.36）个月]和低PLR[（53.6±3.75） vs （37.6±2.96）个月]患者生存时间均长于高NLR和PLR的ICCA患者（均P<0.05）。结论：ICCA术后3 d的NLR和PLR异常增高是患者肝切除术后死亡的独立危险因素，其对患者生存时间具有早期预测价值。

Objective:To observe the clinical characteristics of patients with familial vitreous amyloidosis (FVA) and the efficacy of vitrectomy (PPV) and the occurrence of complications.Methods:A retrospective clinical study. From June 2009 to March 2020, 32 eyes of 18 patients from 3 FVA families who were diagnosed and treated by PPV at Department of Ophthalmology of Jiaxing TCM Hospital were included in the study. Among them, there were 12 males with 22 eyes and 6 females with 10 eyes. The average age of onset was 42.28±3.25 years; the average duration of disease was 3.75±3.93 years. All the affected eyes underwent best corrected visual acuity (BCVA) and B-mode ultrasound examination. A logarithmic visual acuity chart was used in the BCVA examination, which was converted to the logarithmic minimum angle of resolution (logMAR) visual acuity when recorded. The average logMAR BCVA of the affected eye was 1.72±0.53; the intraocular pressure was less than 21 mm Hg (1 mm Hg=0.133 kPa). The vitreous body of the affected eye was obviously cloudy. All the affected eyes underwent standard three-channel PPV through the flat part of the ciliary body, and vitreous specimens were collected for pathological examination during the operation. Peripheral venous blood of probands from 3 families was collected, and the whole exome gene sequencing was performed. The follow-up time after surgery was ≥6 months. The patient's clinical characteristics, fundus lesions in PPV, changes in BCVA after surgery, and complications was observed. One-way analysis of variance or t test was performed for measurement data comparison; χ2 test was performed for count data comparison. Results:The vitreous body of the affected eye showed gray-white dense and thick flocculent changes, and the posterior capsule attached to the lens showed "foot disc-like" turbidity; later the lens was mainly cystic opacity. Pathological examination of the vitreous body showed positive staining of Congo red; under a polarized light microscope, it showed apple green dots and sheet-like birefringence. The genetic test results showed that there was a c.307G>C (p.Gly103Arg) missense mutation in the TTR gene of the proband in Family 2. Peripheral retinal hemorrhages in 4 eyes (12.5%, 4/32), retinal tears in 5 eyes (15.6%, 5/32), retinal degeneration in 4 eyes (12.5%, 4/32), retinal detachment were found in PPV 3 eyes (9.4%, 3/32). The vitreous body was filled with C 3F 8 and silicone oil respectively for 2, 1 eye. Six months after the operation, the logMAR BCVA of the affected eye was 0.39±0.32, which was significantly higher than that before the operation, and the difference was statistically significant ( t=15.131, P=0.000). After the operation, high intraocular pressure occurred in 2 eyes (6.3%, 2/32), secondary glaucoma in 1 eye (3.1%, 1/32), retinal detachment in 2 eyes (6.3%, 2/32), neovascular glaucoma (NVG) in 2 eyes (6.3%, 2/32), cataract in 10 eyes (31.3%, 10/32). Conclusion:The vitreous body of FVA eyes are gray-white dense, thick and flocculent, attached to the posterior lens capsule, showing "foot disc-like" turbidity; PPV treatment can effectively improve the BCVA of the FVA eyes; secondary glaucoma, secondary retinal detachment, NVG can occur after surgery.