Mechanism of Astragaloside Ⅳ on db/db Mice with Type 2 Diabetes Mellitus and Non-alcoholic Fatty Liver Disease Based on AMPK Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the mechanism of astragaloside Ⅳ （AS Ⅳ） on db/db mice with type 2 diabetes mellitus （T2DM） and non-alcoholic fatty liver disease （NAFLD） based on network pharmacology and experimental validation. MethodA total of 24 db/db mice were randomly divided into four groups： model group， metformin group， and low-dose and high-dose AS Ⅳ groups. Six C57 mice were used as the blank group. The low-dose and high-dose AS Ⅳ groups were given AS Ⅳ of 0.015 and 0.030 g·kg-1 by gavage， and the metformin group was given 0.067 g·kg-1 by gavage. The blank and model groups were given equal volumes of distilled water by gavage. After intragastric administration， fasting blood glucose （FBG） was detected， and an oral glucose tolerance test was performed. Serum lipid level and liver histopathology were detected. The target and enrichment pathway of AS Ⅳ for treating T2DM and NAFLD were predicted by network pharmacology， and the main enrichment pathway was verified by molecular biology techniques. The protein expressions of AMPK， p-AMPK， sterol regulatory element-binding protein-1 （SREBP-1）， and fatty acid synthetase （FAS） in liver tissue were detected by Western blot. ResultCompared with the blank group， the levels of body mass， liver weight coefficient， fasting blood glucose， serum total cholesterol， triglyceride， and low-density lipoprotein cholesterol in mice treated with AS Ⅳ were decreased （P<0.05， P<0.01）. The pathology of liver tissue showed significant improvement in lipid accumulation， and imaging results showed that the degree of fatty liver was reduced after AS Ⅳ therapy. Network pharmacological prediction results showed that vascular endothelial growth factor α （VEGFA）， galactoagglutinin 3 （LGALS3）， serine/threonine kinase B2 （Akt2）， RHO-associated coiled-coil protein kinase 1 （ROCK1）， serine/threonine kinase B1 （Akt1）， signaling and transcriptional activator protein （STAT3）， and messtimal epidermal transformation factor （MET） were key targets in "drug-disease" network. The results from the Kyoto encyclopedia of genes and genomes （KEGG） enrichment showed that the AMP-dependent protein kinase （AMPK） signaling pathway was strongly associated with T2DM and NAFLD. Western blot results showed that compared with the blank group， the expression levels of p-AMPK/AMPK in the model group were significantly down-regulated， while those of SREBP-1 and FAS proteins were significantly up-regulated （P<0.01）. Compared with the model group， the expression levels of p-AMPK/AMPK in the metformin group and high-dose AS Ⅳ group were significantly up-regulated， while those of SREBP-1 and FAS proteins were significantly down-regulated （P<0.05， P<0.01）. ConclusionAS Ⅳ regulates the expression of lipid proteins by activating the AMPK signaling pathway， thereby improving lipid metabolism.

Suxiao Jiuxin Pills Prevent Ventricular Fibrillation from Inhibiting L-type Calcium Currents CaV1.2 in vivo and in vitro / 中国结合医学杂志

OBJECTIVE@#To investigate whether Suxiao Jiuxin Pills (SJP), a Chinese herbal remedy, is an anti-ventricular fibrillation (VF) agent.@*METHODS@#VF was induced by isoproterenolol (ISO) intraperitoneal injection followed by electrical pacing in mice and rabbits. The effects of SJP on the L-type calcium channel current (CaV1.2), voltage-dependent sodium channel current (INa), rapid and slow delayed rectifier potassium channel current (IKr and IKs, respectively) were studied by whole-cell patch-clamp method. Computer simulation was implemented to incorporate the experimental data of SJP effects on the CaV1.2 current into the action potential (AP) and pseudo-electrocardiography (pseudo-ECG) models.@*RESULTS@#SJP prevented VF induction and reduced VF durations significantly in mice and rabbits. Patch-clamp experiments revealed that SJP decreased the peak amplitude of the CaV1.2 current with a half maximal concentration (IC50) value of 16.9 mg/L (SJP-30 mg/L, -32.8 ± 6.1 pA; Verapamil, -16.2 ±1.8 pA; vs. control, -234.5 ±16.7 pA, P<0.01, respectively). The steady-state activation curve, inactivation curve, and the recovery from inactivation of the CaV1.2 current were not shifted significantly. Specifically, SJP did not altered INa, IKr, and IKs currents significantly (SJP vs. control, P>0.05). Computer simulation showed that SJP-reduced CaV1.2 current shortened the AP duration, transiting VF into sinus rhythm in pseudo-ECG.@*CONCLUSION@#SJP reduced VF via inhibiting the CaV1.2 current with in vivo, in vitro, and in silico studies, which provide experimental basis for SJP anti-VF clinical application.

The mechanism of S100A7 inducing the migration and invasion in cervical cancer cells / 中华肿瘤杂志

Objective: To investigate the mechanism of S100A7 inducing the migration and invasion in cervical cancers. Methods: Tissue samples of 5 cases of cervical squamous cell carcinoma and 3 cases of adenocarcinoma were collected from May 2007 to December 2007 in the Department of Gynecology of the Affiliated Hospital of Qingdao University. Immunohistochemistry was performed to evaluate the expression of S100A7 in cervical carcinoma tissues. S100A7-overexpressing HeLa and C33A cells were established with lentiviral systems as the experimental group. Immunofluorescence assay was performed to observe the cell morphology. Transwell assay was taken to detect the effect of S100A7-overexpression on the migration and invasion of cervical cancer cells. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to examine the mRNA expressions of E-cadherin, N-cadherin, vimentin and fibronectin. The expression of extracellular S100A7 in conditioned medium of cervical cancer cell was detected by western blot. Conditioned medium was added into Transwell lower compartment to detect cell motility. Exosomes were isolated and extracted from the culture supernatant of cervical cancer cell, the expressions of S100A7, CD81 and TSG101 were detected by western blot. Transwell assay was taken to detect the effect of exosomes on the migration and invasion of cervical cancer cells. Results: S100A7 expression was positively expressed in cervical squamous carcinoma and negative expression in adenocarcinoma. Stable S100A7-overexpressing HeLa and C33A cells were successfully constructed. C33A cells in the experimental group were spindle shaped while those in the control group tended to be polygonal epithelioid cells. The number of S100A7-overexpressed HeLa cells passing through the Transwell membrane assay was increased significantly in migration and invasion assay (152.00±39.22 vs 105.13±15.75, P<0.05; 115.38±34.57 vs 79.50±13.68, P<0.05). RT-qPCR indicated that the mRNA expressions of E-cadherin in S100A7-overexpressed HeLa and C33A cells decreased (P<0.05) while the mRNA expressions of N-cadherin and fibronectin in HeLa cells and fibronectin in C33A cells increased (P<0.05). Western blot showed that extracellular S100A7 was detected in culture supernatant of cervical cancer cells. HeLa cells of the experimental group passing through transwell membrane in migration and invasion assays were increased significantly (192.60±24.41 vs 98.80±47.24, P<0.05; 105.40±27.38 vs 84.50±13.51, P<0.05) when the conditional medium was added into the lower compartment of Transwell. Exosomes from C33A cell culture supernatant were extracted successfully, and S100A7 expression was positive. The number of transmembrane C33A cells incubated with exosomes extracted from cells of the experimental group was increased significantly (251.00±49.82 vs 143.00±30.85, P<0.05; 524.60±52.74 vs 389.00±63.23, P<0.05). Conclusion: S100A7 may promote the migration and invasion of cervical cancer cells by epithelial-mesenchymal transition and exosome secretion.

Effect of Liangxue Huoxue decoction on intestinal flora and NLRP3/caspase-1/GSDMD signaling pathway in mice model of sepsis-induced acute kidney injury / 中华危重病急救医学

Objective:To investigate the effect of Liangxue Huoxue decoction on intestinal flora, intestinal barrier and NOD-like receptor protein 3 (NLRP3)/caspase-1/gasdermin D (GSDMD) pyroptosis signaling pathway in mice model of sepsis-induced acute kidney injury (AKI).Methods:The model of AKI was established by cecal ligation and perforation (CLP). Thirty male C57BL/6 mice were randomly divided into sham operation group (Sham group), sepsis group (CLP group) and sepsis+Liangxue Huoxue decoction (CLP+LXHX group), with 10 mice in each group. Mice in Sham group only underwent laparotomy. Two hours before model establishment, mice in CLP+LXHX group were treated with Liangxue Huoxue decoction (6 g/kg) by gavage; mice in Sham group and CLP group were given equal volume of normal saline by gavages. After 24 hours of modeling, all mice were sacrificed under anesthesia, and the colon and kidney tissues and fresh feces in the colon were taken. The pathological changes of kidney and colon were observed by hematoxylin-eosin (HE) staining under light microscope. Real-time polymerase chain reaction (RT-PCR) was used to detect inflammatory factors (interleukins, IL-1β and IL-18) in renal tissue. The expressions of NLRP3, caspase-1 and GSDMD were detected by Western blotting. The changes of intestinal flora in mice were detected by 16S rDNA high-throughput sequencing.Results:Compared with the Sham group, the inflammatory cell infiltration of the kidney tissue was increased and the kidney became vacuolated in CLP group, the mRNA expressions of IL-1β, IL-18, and the protein expressions of NLRP3, caspase-1 and GSDMD were significantly increased in CLP group, the species richness of intestinal microflora decreased significantly, the relative abundance of Enterococcus and Escherichia-Shigella increased significantly, and the relative abundance of Ileibacterium, Alloprevotella, Lachnospiraceae, Klebsiella and Parasutterella increased significantly in CLP group. Compared with CLP group, Liangxue Huoxue decoction can significantly reduce the pathological changes of kidney and colon tissue, reduce the pathological score (1.75±0.43 vs. 3.50±0.50 for kidney tissue, 1.25±0.43 vs. 4.50±0.50 for colon tissue, both P < 0.05), improve the composition of intestinal flora, reduce the relative abundance of Enterococcus and Escherichia-Shigella, and significantly increase the relative abundance of Lactobacillus and Akkermansia. In addition, Liangxue Huoxue decoction can significantly reduce mRNA expressions of IL-1β and IL-18 in kidney tissue [IL-1β mRNA (2 -ΔΔCt): 1.59±0.05 vs. 4.61±0.88, IL-18 mRNA (2 -ΔΔCt): 1.69±0.17 vs. 2.86±0.63, both P < 0.05] and the protein expressions of NLRP3, caspase-1 and GSDMD (NLRP3/GAPDH: 0.71±0.04 vs. 0.89±0.01, caspase-1/GAPDH: 1.04±0.04 vs. 1.48±0.04, GSDMD/GAPDH: 0.90±0.01 vs. 1.41±0.02, all P < 0.05). Conclusions:Liangxue Huoxue decoction has obvious protective effect on AKI induced by sepsis. It can improve intestinal barrier by regulating intestinal flora, thereby inhibiting the activation of NLRP3/caspase-1/GSDMD signaling pathway in kidney tissue and reducing the expression of proptosis-related inflammatory factors.

Reevaluation of systematic evaluation of Tripterygium glycosides in the treatment of diabetic kidney disease / 中国药房

OBJECTIVE To systematically reevaluate （umbrella review） the systematic review/meta-analysis of Tripterygium glycosides （TG） in the treatment of diabetic kidney disease （DKD）， in order to provide a higher quality evidence-based reference for TG in the treatment of DKD. METHODS The systematic reviews/meta-analysis of TG in the treatment of DKD were searched from CNKI， Wanfang， VIP， CBM， PubMed， Cochrane Library and Embase. The PRISMA 2020 statement， the AMSTAR 2 scale and the GRADE tool were used to evaluate the quality of the report， the quality of the methodology， and the quality of the evidence， respectively. The quantitative results of the included systematic review/meta-analysis were analyzed comprehensively. RESULTS A total of 18 systematic reviews/meta-analyses were included. PRISMA 2020 stated that 3 reports were complete， 13 reports had partial information defects， and 2 reports had serious information defects. The results of the AMSTAR 2 scale evaluation showed that 4 literature had low methodological quality， and 14 literature had very low methodological quality. GRADE tool evaluation results showed that there were 106 outcome indicators， including 34 intermediate-quality evidence accounted for 32.1%， 51 poor-quality evidence accounted for 48.1%， 21 very poor-quality evidence accounted for 19.8%， and there was no high- quality evidence. Comprehensive analysis of quantitative results of various outcome indicators showed that TG had definite improvement effects on the total effective rate of DKD， 24-hour urinary protein quantity and serum albumin， and the adverse drug reactions were different in every study. CONCLUSIONS The efficacy of TG in the treatment of DKD is relatively accurate， safety still needs to be paid attention to， and future studies with larger sample size need to be verified.

Mid term results of fenestrated branch stent technique for the treatment of thoracoabdominal aortic aneurysms / 中华普通外科杂志

Objective:To evaluate the mid-term results of fenestrated/branched endovascular aortic repair (f/b EVAR) for the treatment of thoracoabdominal aortic aneurysms. M ethods The clinical data of 105 thoracoabdominal aortic aneurysm patients treated with f/b EVAR at the Department of Vascular Surgery of Nanjing Drum Tower Hospital from 2018 to 2019 were retrospectively analyzed. Results:There were 43 cases of thoracoabdominal aortic aneurysm and 62 cases of thoracoabdominal aortic aissection.A total of 336 branch arteries were reconstructed,and technical success rate was 94.3%. 100 cases (95.2%) were followed-up, 6 cases (5.7%) received reoperation interventions, and 11 cases (10.5%) died. During the follow-up period, 69 cases had complete imaging data. Based on the recent CT date of the thoracoabdominal aorta, 58 patients hael positive aortic remodeling and 11 patients hael negative and indeterminate remodeling; there were 31 cases (29.5%) of endoleaks, including 7 cases (6.7%) of type Ⅰb endoleaks, 8 cases (7.6%) of type Ⅱ, 1 case (0.95%) of type Ⅲa, 13 cases (12.4%) of type Ⅲc endoleaks and 2 cases (1.9%) of type Ⅳ. Conclusions:The mid-term follow-up results were satisfactory for TAAA treated with f/b EVAR. Internal leakage remains key point for f/b EVAR.

Analysis of clinical characteristics of 36 patients with pyoderma gangrenosum and comparison between the PARACELSUS score and Delphi criteria for their diagnosis / 中华皮肤科杂志

Objective:To analyze clinical characteristics, treatment, and outcomes of 36 patients with pyoderma gangrenosum, and to compare and evaluate the applicability and consistency between the PARACELSUS score and Delphi criteria.Methods:From January 2000 to January 2022, clinical data were collected from 36 patients who were diagnosed with pyoderma gangrenosum in the Hospital of Dermatology, Chinese Academy of Medical Sciences. The PARACELSUS score and Delphi criteria were applied to their diagnosis, and the kappa test was used to evaluate the consistency between the two diagnostic criteria.Results:Among the 36 patients, 6 (16.67%) had definite precipitating factors before the onset, and 31 (86.11%) exhibited lesions with different degrees of pain. Ulcerative lesions predominatd in 31 (86.11%) patients, which mostly involved the lower extremities, while 16 (44.44%) presented with multiple lesions all over the body. Four (11.11%) patients were complicated by inflammatory bowel disease, and 3 (8.33%) with inflammatory arthritis. Glucocorticoids, Tripterygium wilfordii, and cyclosporin were the main systemic treatment options, and tumor necrosis factor-α antagonist was used in 8 (22.22%) patients. Twenty-two (64.71%) and 17 (50.00%) patients were diagnosed with pyoderma gangrenosum using the PARACELSUS score and Delphi criteria, respectively. The kappa test showed moderate agreement between the two diagnostic criteria (kappa value = 0.47, 95% CI: 0.19 - 0.76, P = 0.004) . Conclusions:Classic ulcerative subtype was the major subtype in the patients with pyoderma gangrenosum, who were usually complicated by inflammatory bowel disease and inflammatory arthritis. Glucocorticoids and immunosuppressants were the main therapeutic drugs. The PARACELSUS score and Delphi criteria focus on different aspects of pyoderma gangrenosum, and the PARACELSUS score is recommended in the context of absence of typical histopathological manifestations.

Clinical analysis of 31 cases of fetal umbilical artery thrombosis / 中华妇产科杂志

Objective: To analyze the ultrasonic manifestations, clinical features, high risk factors and key points of pregnancy management in prenatal diagnosis of umbilical artery thrombosis (UAT). Methods: The data of 31 pregnant women of UAT diagnosed by prenatal ultrasonography and confirmed after birth from July 2017 to July 2022 at the Women's Hospital, Zhejiang University School of Medicine were retrospectively analyzed, including the maternal characteristics, pregnancy outcomes and fetal complications. In addition, the baseline data and pregnancy outcomes were compared in 21 patients who continued pregnancy after diagnosis of UAT. Of the 21 UAT cases that continued pregnancy, 10 cases were treated with low molecular weight heparin (LMWH; LMWH treatment group), while the other 11 patients had expectant treatment(expectant treatment group). Results: The age of the 31 pregnant women was (30.2±4.7) years, of which 5 cases (16%,5/31) were advanced age pregnant women. The gestational age at diagnosis was (32.9±4.0) weeks, and the gestational age at termination of pregnancy was (35.6±2.9) weeks. In 31 fetuses with UAT, 15 cases (48%) had fetal distress, 11 cases (35%) had fetal growth restriction, and 3 cases (10%) had intrauterine stillbirth. There were 28 cases of live births, including 26 cases by cesarean section and 2 cases by vaginal delivery. There were also 3 stillbirths, all delivered vaginally. Four neonates had mild asphyxia and two newborns had severe asphyxia. Among the 31 cases, 10 cases were terminated immediately after diagnosis, the gestational age at diagnosis was (35.9±2.9) weeks. Another 21 pregnancies continued, and their gestational age at diagnosis was (31.4±3.7) weeks. The median prolonged gestational age in LMWH treatment group was 7.9 weeks (4.6-9.4 weeks), and all were live births. The median prolonged gestational age in the expectant treatment group was 0.6 weeks (0.0-1.0 weeks), and 2 cases were stillbirths. There was a statistically significant difference in prolonged gestational age (P=0.002). Conclusions: Ultrasound is the preferred method for prenatal detection of UAT. Clinicians need to be vigilant for UAT when a newly identified single umbilical artery is detected by ultrasound in the second or third trimesters. The decision to continue or terminate the pregnancy depends on the gestational age and the condition of fetus. Attention should be paid to fetal movements as the pregnancy continues. The treatment of LMWH as soon as possible after diagnosis of UAT may improve the pregnancy outcome.

Zuogui Jiangtang Qinggan Prescription promotes recovery of intestinal mucosal barrier in mice with type 2 diabetes mellitus and nonalcoholic fatty liver disease by improving intestinal flora homeostasis / 中国中药杂志

This study aimed to investigate the recovery effect of Zuogui Jiangtang Qinggan Prescription on intestinal flora homeostasis control and intestinal mucosal barrier in type 2 diabetes mellitus(T2DM) with nonalcoholic fatty liver disease(NAFLD) induced by a high-fat diet. NAFLD was established in MKR transgenic mice(T2DM mice) by a high-fat diet(HFD), and subsequently treated for 8 weeks with Zuogui Jiangtang Qinggan Prescription(7.5, 15 g·kg~(-1)) and metformin(0.067 g·kg~(-1)). Triglyceride and liver function were assessed using serum. The hematoxylin-eosin(HE) staining and Masson staining were used to stain the liver tissue, while HE staining and AB-PAS staining were used to stain the intestine tissue. 16S rRNA sequencing was utilized to track the changes in the intestinal flora of the mice in each group. Polymerase chain reaction(PCR) and immunofluorescence were used to determine the protein and mRNA expression levels of ZO-1, Occludin, and Claudin-1. The results demonstrated that Zuogui Jiangtang Qinggan Prescription increased the body mass of T2DM mice with NAFLD and decreased the hepatic index. It down-regulated the serum biomarkers of liver function and dyslipidemia such as alanine aminotransferase(ALT), aspartate transaminase(AST), and triglycerides(TG), increased insulin sensitivity, and improved glucose tolerance. According to the results of 16S rRNA sequencing, the Zuogui Jiangtang Qinggan Prescription altered the composition and abundance of the intestinal flora, increasing the relative abundances of Muribaculaceae, Lactobacillaceae, Lactobacillus, Akkermansia, and Bacteroidota and decreasing the relative abundances of Lachnospiraceae, Firmicutes, Deslfobacteria, Proteobacteria, and Desulfovibrionaceae. According to the pathological examination of the intestinal mucosa, Zuogui Jiangtang Qinggan Prescritpion increased the expression levels of the tight junction proteins ZO-1, Occludin, and Claudin-1, promoted intestinal mucosa repair, protected intestinal villi, and increased the height of intestinal mucosa villi and the number of goblet cells. By enhancing intestinal mucosal barrier repair and controlling intestinal microbiota homeostasis, Zuogui Jiangtang Qinggan Prescription reduces intestinal mucosal damage induced by T2DM and NAFLD.

Analysis of the effects of low/intermediate dose of coagulation factor Ⅷ on 30 adult patients with severe hemophilia A in a single center / 中华血液学杂志

Objective: To evaluate the clinical effects of low- and intermediate-dose factor Ⅷ (F Ⅷ) prophylaxis in Chinese adult patients with severe hemophilia A. Methods: Thirty adult patients with severe hemophilia A who received low- (n=20) /intermediate-dose (n=10) F Ⅷ prophylaxis at Nanjing Drum Tower Hospital affiliated with Nanjing University Medical College were included in the study. The annual bleeding rate (ABR), annual joint bleeding rate (AJBR), number of target joints, functional independence score of hemophilia (FISH), quality of life score, and health status score (SF-36) before and after preventive treatment were retrospectively analyzed and compared. Results: The median follow-up was 48 months. Compared with on-demand treatment, low- and intermediate-dose prophylaxis significantly reduced ABR, AJBR, and the number of target joints (P<0.05) ; the improvement in the intermediate-dose prophylaxis group was better than that in the low-dose prophylaxis group (P<0.05). Compared with on-demand treatment, the FISH score, quality of life score, and SF-36 score significantly improved in both groups (P<0.05), but there was no significant difference between the two groups (P>0.05) . Conclusion: In Chinese adults with severe hemophilia A, low- and intermediate-dose prophylaxis can significantly reduce bleeding frequency, delay the progression of joint lesions, and improve the quality of life of patients as compared with on-demand treatment. The improvement in clinical bleeding was better with intermediate-dose prophylaxis than low-dose prophylaxis.

Shexiang Tongxin Dropping Pill Allieviates Heart Failure via Extracellula Matrix-Receptor Interaction Pathways Based on RNA-Seq Transcriptomics and Experimental Studies / 中国结合医学杂志

OBJECTIVE@#To investigate the protective mechanisms of Chinese medicine Shexiang Tongxin Dropping Pills (STDP) on heart failure (HF).@*METHODS@#Isoproterenol (ISO)-induced HF rat model and angiotensin II (Ang II)-induced neonatal rat cardiac fibroblast (CFs) model were used in the present study. HF rats were treated with and without STDP (3 g/kg). RNA-seq was performed to identify differentially expressed genes (DEGs). Cardiac function was evaluated by echocardiography. Hematoxylin and eosin and Masson's stainings were taken to assess cardiac fibrosis. The levels of collagen I (Col I) and collagen III (Col III) were detected by immunohistochemical staining. CCK8 kit and transwell assay were implemented to test the CFs' proliferative and migratory activity, respectively. The protein expressions of α-smooth muscle actin (α-SMA), matrix metalloproteinase-2 (MMP-2), MMP-9, Col I, and Col III were detected by Western blotting.@*RESULTS@#The results of RNA-seq analysis showed that STDP exerted its pharmacological effects on HF via multiple signaling pathways, such as the extracellular matrix (ECM)-receptor interaction, cell cycle, and B cell receptor interaction. Results from in vivo experiments demonstrated that STDP treatment reversed declines in cardiac function, inhibiting myocardial fibrosis, and reversing increases in Col I and Col III expression levels in the hearts of HF rats. Moreover, STDP (6, 9 mg/mL) inhibited the proliferation and migration of CFs exposed to Ang II in vitro (P<0.05). The activation of collagen synthesis and myofibroblast generation were markedly suppressed by STDP, also the synthesis of MMP-2 and MMP-9, as well as ECM components Col I, Col III, and α-SMA were decreased in Ang II-induced neonatal rats' CFs.@*CONCLUSIONS@#STDP had anti-fibrotic effects in HF, which might be caused by the modulation of ECM-receptor interaction pathways. Through the management of cardiac fibrosis, STDP may be a compelling candidate for improving prognosis of HF.

Study of the effects of long-term outcomes of autologous peripheral blood stem cell reinfusion in patients with decompensated cirrhosis / 中华肝脏病杂志

Objective: Autologous peripheral blood stem cells (PBSC) derived from bone marrow can promote liver regeneration and improve the liver function of patients, but there are few studies on its effect on the long-term outcomes in patients with decompensated cirrhosis. Based on previous work, this study observed the clinical outcomes of PBSC treatment in patients with decompensated cirrhosis for 10 years, in order to provide more data support for the safety and efficacy of stem cells in clinical applications. Methods: Data of patients with decompensated liver cirrhosis who completed PBSC treatment in the Department of Gastroenterology of the First Affiliated Hospital of Air Force Military Medical University from August 2005 to February 2012 were included. The follow-up endpoint was death or liver transplantation, and patients who did not reach the follow-up endpoint were followed-up for at least 10 years. The patients with decompensated liver cirrhosis who met the conditions for PBSC treatment but did not receive PBSC treatment in our hospital during the same period were used as controls. Results: A total of 287 cases with decompensated liver cirrhosis had completed PBSC treatment, and 90 cases were lost to follow-up within 10 years after surgery. A total of 151 cases with complete survival follow-up data were included in the control group. There were no statistically significant differences in baseline information such as gender, age, etiological composition and liver function score between the two groups. The 10-year survival rate was higher in PBSC than control group (37.56% vs. 26.49%, P<0.05). Cholinesterase, albumin, international normalized ratio, Child-Turcotte-Pugh score, model for end-stage liver disease score, and other indicators were gradually recovered within 3 months to 1 year after PBSC treatment, and stabilized at a more desirable level in the long-term after follow-up for up to 10 years. There was no statistically significant difference in the incidence of liver cancer between the two groups (25.22% vs.31.85%, P=0.267). The age of onset of hepatocellular carcinoma was later in PBSC than control group [(56.66±7.21) years vs. (52.69±8.42) years, P<0.05]. Conclusions: This long-term observational follow-up study of more than ten years confirms that PBSC treatment can bring long-term benefits to patients with decompensated cirrhosis, with good long-term safety, thus providing more data support on the safety and efficacy of stem cells for clinical applications.

Research on the mechanism of bufalin inhibiting migration and invasion of human prostate cancer PC3 cells / 药学学报

In this study, we investigated the inhibitory effects of bufalin on proliferation, migration and invasion of PC3 cells in vitro, and preliminarily explored the molecular mechanism of epithelial-mesenchymal transformation (EMT) inhibited by bufalin. The viability of PC3 cells was evaluated by MTT assay, and the migration and invasion abilities of PC3 cells were detected by wound healing and Transwell assay. Western blot was used to detect the expression of EMT and integrin family proteins. The results showed that the half maximal inhibitory concentration (IC50) value of bufalin against PC3 cells was 0.26 ± 0.03 μmol·L-1. After bufalin treatment, the migration rate of PC3 cells slowed down (P < 0.05), the number of PC3 cells passing through the microporous membrane decreased (P < 0.05), which indicated that bufalin could inhibit the proliferation, migration and invasion of PC3 cells in a concentration-dependent manner. We found that bufalin could affect the expression of EMT-related proteins,including up-regulation of E-cadherin and down-regulation of N-cadherin, β-catenin, matrix metalloproteinase 9 (MMP9), matrix metalloproteinase 2 (MMP2), c-myc and Snail. Bufalin also inhibited the expression of integrin family proteins, including integrin α2 (ITGA2), integrin β1 (ITGB1), integrin β3 (ITGB3), integrin β5 (ITGB5), Yes-associated protein/transcriptional coactivator with a PDZ-binding motif (YAP/TAZ) and integrin-linked kinase (ILK). In addition, bufalin could also inhibit the protein expression level of phospho-focal adhesion kinase (p-FAK)/FAK, phospho-steroid receptor coactivator (p-Src)/Src and phospho-protein kinase B (p-Akt)/Akt. These results suggested that bufalin might inhibit the proliferation, metastasis and invasion of prostate cancer PC3 cells through the FAK/Src/phosphoinositide 3-kinase (PI3K)/Akt pathway. Therefore, bufalin provides reference value for the development of therapeutic drugs for prostate cancer.

Risk factors analysis of central airway stenosis after lung transplantation / 器官移植

Objective To investigate the risk factors of central airway stenosis after lung transplantation. Methods Clinical data of 155 recipients undergoing lung transplantation in Wuxi People's Hospital Affiliated to Nanjing Medical University from July 2016 to December 2017 were retrospectively analyzed. According to the incidence of central airway stenosis following lung transplantation, all recipients were divided into the stenosis group (n=36) and control group (n=119). The incidence of central airway stenosis after lung transplantation was summarized. The risk factors of central airway stenosis after lung transplantation were assessed by univariate and multivariate logistic regression analyses. Results Among 155 lung transplant recipients, 36 cases (23.2%) developed central airway stenosis. The average incidence time was (53±13) d after lung transplantation. Univariate analysis demonstrated that bilateral lung transplantation, grade 3 primary graft dysfunction (PGD), airway fungal infection, long cold ischemia time, long mechanical ventilation time and long intensive care unit (ICU) stay were the risk factors for central airway stenosis after lung transplantation (all P < 0.05). Multivariate analysis showed that airway fungal infection, long cold ischemia time and long mechanical ventilation time were the independent risk factors for central airway stenosis after lung transplantation (all P < 0.05). Conclusions Airway fungal infection after lung transplantation, long cold ischemia time and long mechanical ventilation time probably lead to central airway stenosis after lung transplantation. Active preventive measures and intimate monitoring should be taken to improve the quality of life of the recipients after lung transplantation.

Role of intercellular junctions in the biological behavior of SP cells of multiple myeloma / 药学实践杂志

Objective To observe the effects of the intercellular gap junction (GJIC) composed of connexin 43(Cx43) in mesenchymal stem cells (MSCs) from different sources and their signals on the biological behavior of multiple myeloma (MM) lateral population cells (SP cells), and to explore its possible mechanism. Methods Mesenchymal stem cells (MSCs) from different sources were isolated and cultured. SP cells of MM cell line RPMI 8266 were sorted by flow cytometry. RT-PCR and Western blot were used to detect the expression of Cx43 gene and protein in MSCs, RPMI 8266 and SP cells from different sources. The effects of MSCs from different sources on SP cell cycle, Cx43 protein expression, colony formation ability in vitro, stem cell related gene expression, cytokine secretion and drug resistance were observed. Results There was no significant difference in morphology and phenotype between MM-MSCs and ND-MSCs. Both MM-MSCs and RPMI 8266 cells expressed a higher level of Cx43. Co-culture with MM-MSCs induced more SP cells to enter G0 phase (P<0.001). The expressions of c-myc, Kif4 and Sox2 genes in SP cells were significantly up-regulated, while the expression of Oct-4 gene was down-regulated. After adding α-GA, c-myc, Kif4 and Sox2 were down-regulated in varying degrees, but there was no significant difference. The expression of Cx43 was up-regulated by (31.00±2)% and (39.00±2)%, respectively. The colony formation ability in vitro was up-regulated, and the addition of α-GA could partially inhibit this effect. A small amount of c-myc, Kif4, Sox2 and Oct-4 genes were expressed in RPMI 8266. These genes were significantly up-regulated in SP cell subpopulation. MM-MSCs secreted high levels of interleukin (IL)-6. After co-culture with SP cells, the expressions of IL-6, IL-10 and TGF-β in the supernatant of MM-MSCs were up-regulated (P=0.0072, P=0.037). bFGF and IL-17 had no significant change. After adding α-GA, the levels of IL-6, IL-10 and TGF-β in the supernatant decreased. MM cells were sensitive to bortezomib (BTZ) induced apoptosis, but SP cells were less sensitive. Co-culture with MM-MSCs significantly reduced BTZ-mediated apoptosis. The addition of α-GA partially restored the sensitivity of MM cells to bortezomib. Conclusion MM-MSCs and multiple myeloma SP cells up-regulate the expression of Cx43 protein, form more GJIC, and promote the proliferation and drug resistance of SP cells by changing the cytokine secretion profile of MSCs, which may be one of the reasons for the recurrence of MM.

Changes of viral DNA load in the early stages of antiviral treatment in varicella zoster virus-induced acute retinal necrosis / 中华眼底病杂志

Objective:To observe the changes of varicella zoster virus (VZV)-DNA load in aqueous humour samples in VZV-induced acute retinal necrosis (ARN) in the early stages of antiviral treatment.Methods:A retrospective observational clinical study. From April 2016 to April 2018, 24 patients with 24 eyes of VZV-induced ARN who were diagnosed by Department of Ophthalmology, Eye and ENT Hospital of Fudan University and received complete aqueous humor sampling were included in the study. Among them, there were 13 males with 13 eyes, 11 females with 11 eyes; 12 left eyes and 12 right eyes; the age was 52.0±9.5 years old (39-71 years old). The time from the onset of ocular symptoms to the diagnosis of ARN was 16.6±6.1 days (7-30 days). Best-corrected visual acuity (BCVA) and ultra-wide-field fundus imaging were performed in all affected eyes. The BCVA examination was carried out using the Snellen visual acuity chart, which was converted into the logarithm of the minimum angle of resolution (logMAR) visual acuity. All patients were given intravitreal injection of 40 mg/ml ganciclovir 0.1 ml (including 4 mg of ganciclovir), 2 times a week, until the active necrotizing retinal lesions subsided, at most after the diagnosis 4 weeks, with a maximum of 9 injections. The follow-up period was 12.8±5.6 months. The aqueous humor samples were collected at presentation and 4, 7, 14, 21, 28 days after the initiation of antiviral therapy, and the VZV-DNA load was detected by real-time quantitative polymerase chain reaction. A plateau phase and a logarithmic reduction phase of the DNA load changes were observed after antiviral treatment began. Wilcoxon rank sum test was used to compare and analyze the differences in BCVA between the eyes at baseline and last follow-up.Results:The mean viral load at presentation was 8.6×10 7±1.3×10 8 copies/ml. The initial plateau phase last for an average of 7.4±2.4 days. In the following logarithmic reduction phase, the mean slope of the decline in viral load was -0.13±0.04 log/day, and the expected time for half reduction of the initial viral load was 2.5±0.7 days. After 28 days antiviral treatment, the viral load decreased to 1.7×10 5±1.8×10 5 copies/ml. In the course of the disease, rhegmatogenous retinal detachment occurred in 16 eyes. Before treatment and at the last follow-up, the logMAR BCVA of the affected eye was 1.1±0.6 and 0.8±0.7, respectively. The results of correlation analysis showed that the logMAR BCVA at the last follow-up was correlated with the initial VZV-DNA load ( r=0.467, P=0.033). Conclusion:The VZV-DNA load in the aqueous humor of eyes with VZV-induced ARN is significantly decreased after antiviral treatment, which is closely related to the clinical process of ARN.

Therapeutic effect of exposed bone cement in treatment of infectious bone and soft tissue defect / 中华显微外科杂志

Objective:To evaluate the therapeutic effect of exposed bone cement in treatment of infectious bone and soft tissue defect by comparison with routine bone cement therapy.Methods:A retrospective analysis was carried out in 27 patients who had been treated from January 2016 to January 2020. Of the 27 patients, 12 were treated by exposed bone cement filling for bone defect at the first stage, followed by flap and bone grafting at the second stage. Other 15 patients were treated by routine bone cement filling and flap repair at the first stage, followed by bone grafting at the second stage. Regular dressing change was carried out after surgery. All patients entered follow-up by out-patient review for wound condition and X-ray. Infection rate of bone and soft tissue, time of bone union, flap survive rate and complication were compared between the 2 groups.Results:The follow-up lased for 9-24(16.5±3.9 ) months. The bone defects were all healed. The treatment time of bone defects in the group of exposed bone cement was 25-34(28.5±2.8) weeks. The treatment time of bone defect in the routine cement group was 25-36(29.6±3.4 )weeks. There was no statistical difference between the 2 groups( P<0.05). Two cases in each group had failed in infection control after bone cement implantation. With further debridement, cleaning and other treatment, the infection was under control. All flaps in the 2 groups survived. In the group of exposed bone cement, there were 3 flaps splitting, 2 hemorrhage(effusion) and 1 partial flap necrosis, In the routine bone cement group, there were 4 wound dehiscence, 8 hemorrhage(effusion) and 3 partial flaps necrosis. There was no significant difference in flap complications between the 2 groups( P>0.05). The healing of bone defect was evaluated by Samantha X-ray score, the scores of the 2 groups were 5.41±0.67 and 5.40±0.63, respectively, with no statistical significance. The Paley's approach was used to grade the bone healing and the function of adjacent joints. Paley bone defect healing evaluation results weve all excellent. Joint function assessment were as follow: in the group of exposed bone cement, 6 cases were excellent and 3 were good; in routine bone cement group: 6 cases were in excellent and 5 in good. There was no significant statistical difference. Conclusion:Compared with a routine bone cement treatment, the exposed bone cement in the treatment of infectious bone defects saw a fewer flap splitting and fewer hemorrhage/effusion, without an increase in bone and soft tissue infection. This procedure could be considered for further trials in the treatment of a composite defects of infected bone and soft tissue.

Analysis of inflammatory characteristics and changes of small airway function in patients with eosinophil and neutrophil asthma / 中国综合临床

Obsjective To analyze the inflammation characteristics and changes of small airway function in patients with eosinophil and neutrophil asthma, and provide evidence for individualized treatment of asthma. Methods:Using a cross-sectional study, 46 patients with eosinophilic asthma and 42 patients with neutrophilic asthma confirmed by cytology of induced sputum were recruited from July 1, 2017 to June 30, 2019 at the respiratory Department of Respiratory Medicine,Jinshan Branch of the Sixth People's Hospital of Shanghai. Patients were divided by asthma category into eosinophilic asthma group and neutrophilic asthma group.The severity of acute attack, the score of asthma control test (ACT) and the concentration of serum C-reactive protein (CRP) were compared between the two groups The fraction of exhaled nitric oxide (FeNO), related cytokines(interleukin-4(IL-4), interleukin-5(IL-5), interleukin-13(IL-13), interleukin-17(IL-17) and interferon γ(IFN-γ)) in peripheral blood and induced sputum supernatant and lung function indicators (forced exhalation volume in one second (FEV1)% percent predicted (%pred), maximal mid-expiratory flow (MMEF)% pred, forced expiratory flow (FEF) 75% pred, forced expiratory flow at 50% of FVC exhaled (FEF50%) pred were detected. Independent sample t-test was used for the comparison between measurement data groups comforming to normal distritution, rank sum test was used for the comparison between measurement data groups not conforming to normal distribution, and χ 2 test was used for the comparison of counting data. Results:There were no significant differences in the general data and ACT scores between the two groups (all P>0.05). The ratio of severe and critical degree (52.38%(22/42)), uncontrolled and partially controlled patients (59.52%(25/42)), CRP level (24.6(7.1, 35.0) mg/L) in neutrophil asthma group were higher than those in eosinophilic asthma group(30.43% (14/46), 36.96% (17/46), and 8.5 (2.0, 12.0) mg/L, respectively) (χ 2=4.37, χ 2=4.48, Z=4.76; P=0.036, P=0.034, P<0.001). The concentration of FeNO was higher in eosinophilic asthma group (76(54,93) ppb) than that in neutrophil asthma group(27(15,41) ppb),and the differences was statistically significant ( Z=6.52, P<0.001). The values of FEV1% pred ((56.13±21.51)%), MMEF% pred ((62.03±23.97)%), FEF75% pred ((54.42±20.49)%), FEF50% pred ((66.89±26.47)%) in neutrophil asthma group were lower than those in eosinophilic asthma group ((68.53±29.81)%, (72.16±23.05)%, (65.38±25.46)% and (79.86±27.61)%), and the difference between the two groups was statistically significant( t values were 2.25, 2.02, 2.21, 2.24; P values were 0.027, 0.046, 0.030, 0.027). The concentrations of serum IL-4((49.42±24.46) ng/L), IL-5((104.89±43.91) ng/L) and IL-4((44.49±19.12) ng/L), IL-5((95.45±28.58) ng/L) in induced sputum supernatant were higher than neutrophilic asthma group((32.29±14.19), (50.35±22.30), (33.33±15.08), (55.61±26.41) ng/L). The difference between the two groups was statistically significant ( t values were 4.06, 7.44, 3.02, 6.77, P values were <0.001, <0.001, 0.003, <0.001). In eosinophilic asthma group, the concentrations of serum IL-13 ((76.18±20.62) ng/L), IL-17 ((31.32±9.32) ng/L), IFN-γ ((18.27±5.56) ng/L) and IL-13((71.08±20.08) ng/L), IL-17((26.29±6.70) ng/L), and IFN-γ((17.61±5.94) ng/L) in induced sputum supernatant were lower than those in neutrophilic asthma group((153.83±44.53 ) ng/L, (55.27±18.89) ng/L, (26.46±10.08) ng/L, (120.32±28.41) ng/L, (44.99±12.66) ng/L, (23.91±7.66) ng/L). The difference between the two groups was statistically significant ( t values were 10.33, 7.43, 4.66,9.31,8.54,4.33, respectively; all P<0.001). Conclusion:Eosinophilic asthma and neutrophil asthma have different inflammation, small airway function characteristics and different response to treatment. The small airway function changes in early stage of neutrophil asthma are more obvious.

Data mining of medical rule in Lingnan TCM masters for treating Allergic rhinitis / 国际中医中药杂志

Objective:To analyze the characteristics of prescription about Lingnan Traditional Chinese Medicine (TCM) masters for Biqiu treatment based on data mining, so as to provide reference for the clinical practice and patent drug research and development of rhinorrhea in Lingnan district.Methods:By searching journal documents and medical records in CNKI, VIP, Wanfang, Chinese BioMedical Database (SinoMed) and Duxiu Data Retrieval Platform, we selected the articles and famous TCM medical records that met the criteria on the TCM medical rules of treating Biqiu. We screened database in strict accordance with inclusion and exclusion criteria and extracted prescription information from medical cases. The software Ancient and Modern Consilia Cloud Platform (V 1.5) was used for data mining analysis and medical case standardization, and SPSS Clementine 12.0 software were used to conduct frequency statistics, association analysis and cluster analysis of drugs in medical cases. Results:A total of 31 articles and 42 medical cases including 73 prescriptions were screened. The frequency results showed that there were 40 kinds of high frequency TCMs (frequency more than 5), mainly including drugs for benefiting the qi and strengthening the spleen, medicine to relieve cold and promote nasal passages, which was the same as the frequency statistics of the effects of herbal medicine. Seven drug groups were obtained by cluster analysis. The results of rule analysis showed that 16 association rules for drug pairs and 18 association rules for 3 TCMs pairs were obtained.Conclusions:While treating allergic rhinitis, Lingnan medical doctors focus on tonifying spleen qi, dispersing wind and cold, and promoting the nasal passages, which reflect the treatment theory of cultivating earth and generating gold, and attacking and tonifying. Lingnan medical doctors commonly used Guangdong medicinal herbs, which have distinct regional characteristics. The data mining methods can comprehensively summarize and explore the potential rules of Lingnan prominent practitioners' treatment of allergic rhinitis, and provide reference for guiding the treatment of congested nose and congested nose in TCM.

External fixation combined with Prontosan management for open fractures complicated with multidrug-resistant bacterial infection / 中华创伤骨科杂志

Objective:To access the efficacy of external fixation combined with Prontosan management for open fractures complicated with multidrug-resistant bacterial infection.Methods:A retrospective analysis was conducted of the data of 22 patients with open fracture complicated by multidrug-resistant bacterial infection who had been admitted to Department of Orthopedic Trauma and Microsurgery, Zhongnan Hospital of Wuhan University from January 2015 to January 2020. According to whether the Prontosan management was used or not, the patients were divided into 2 groups.The Protosan group [9 males and 3 females with an age of (44.6±13.1) years] were subjected to external fixation, vacuum sealing drainage(VSD) or conventional dressing changes, and at the same time Protosan management to flush the wound or Prontosan gel to change dressings.The control group[6 males and 4 females with an age of (45.1±11.7) years] were subjected to external fixation plus VSD or conventional dressing changes. Skin flaps or skin grafts were used to repair the wound immediately after the wound infection was controlled in both groups. The time for negative culture of the bacteria on the wound surface, number of debridement, survival of the skinflaps or grafts, and fracture union were recorded and compared between the 2 groups.Results:There was no significant difference in the preoperative general data between the 2 groups which were comparable( P>0.05).All patients were followed up for 10 to 24 months (mean, 14.5 months).In the Prontosan group, the number of debridement was 2.0 (2.0, 3.0) times, significantly fewer than that in the control group [4.0 (3.0, 4.0) times]; the time for negative bacterial culture (8.3±2.2) d, significantly shorter than that in the control group [(14.2±3.1) d]; the fracture union time (5.5±1.1) months, significantly shorter than that in the control group [(6.5 ±1.1) months]; the Samantha X-ray score at 6 months after operation 6.0 (5.0,6.0) points, significantly higher than that in the control group [5.2(4.5,5.5) points] (all P<0.05). Skin flaps or grafts survived in all the patients without any nonunion or chronic osteomyelitis. Conclusion:In the external fixation of open fractures complicated with multidrug-resistant bacterial infection, combination with Prontosan management can effectively control infection and promote granulation, early wound healing and fracture union.