UPLC-MS/MS Method for Detection of Etomidate and Its Metabolite Etomidate Acid Quantity in Blood / 法医学杂志

OBJECTIVES@#To establish a method for the simultaneous quantitative analysis of etomidate and its metabolite etomidate acid in blood, and to discuss its application value in actual cases.@*METHODS@#Acetonitrile precipitate protein method was used, and C18 column was selected. Gradient elution was performed with acetonitrile and 5 mmol/L ammonium acetate within 6 min. Electrospray ionization source in positive ion mode was used. The internal standard etomidate acid-d5 was obtained by etomidate-d5 alkaline hydrolysis reaction. Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for quantitative analysis. The methodological verification was conducted.@*RESULTS@#Etomidate and etomidate acid in blood showed good linear relationship in the quantitative linear range (r>0.999), with the lower limit of quantification was 2.5 ng/mL and 7.5 ng/mL, respectively. The accuracy, precision, recovery rate, and matrix effect of the method met the professional verification standards. The practical application results showed that etomidate and etomidate acid could be detected in the blood of the abusers, and their mass concentrations ranged from 17.24 to 379.93 ng/mL.@*CONCLUSIONS@#The method established in this study can simultaneously quantify etomidate and etomidate acid in blood, which is simple and convenient to operate with accuracy. It can meet the detection needs of actual cases and provide technical support for law enforcement to crack down on etomidate abuse.

Application and value of intravascular ultrasound for excimer laser ablation combined with drug-coated balloon in the treatment of lower limb arteriosclerotic obliterans / 中华外科杂志

Objective: To examine the value of intravascular ultrasound (IVUS) for excimer laser ablation (ELA) combined with drug-coated balloon (DCB) in treating lower limb arteriosclerotic obliterans (ASO). Methods: As a prospective case series study, patients who underwent ELA combined with DCB for lower limb ASO with the guidance of IVUS from September 2021 to March 2022 at Department of Vascular Surgery, Zhongshan Hospital, Fudan University were enrolled prospectively. Lesion characteristics, procedure-related outcomes and complications were collected. The therapy outcomes were compared with baseline data by paired t test. Results: There were 8 males and 2 females, aged (72.0±5.9) years (range: 61 to 81 years). Of all the 11 lesions, there were 8 lesions in superficial femoral artery and 3 in popliteal artery. The lesion length was (7.0±2.4) cm (range: 3.2 to 9.8 cm). There were 4 chronic totally occlusion and 7 severe stenosis. All patients underwent the operation successfully. The technical success rate was 10/11. Bailout stenting was performed in one lesion because of flow-limiting dissection. Four lesions were grade 3 to 4 in peripheral artery calcium score system, and 9 lesions with calcification arc≥180°. Larger diameter drug-coated balloons were selected in 5 lesions after measurement of intravascular ultrasound. The follow-up time was (6.0±1.9) months (range: 3 to 9 months). The ankle-brachial index of the patient was significantly improved immediately after surgery (0.97±0.13 vs. 0.48±0.18, t=-7.60, P<0.01) and at 3 months after surgery (0.95±0.12 vs. 0.48±0.18, t=-7.17, P<0.01). The 3-month primary patency rate was 11/11, the target lesion reintervention was 0 and ulcer healing rate was 3/4. Conclusion: IVUS assisted ELA in the treatment of lower limb artery lesions is safe and effective in early stage.

Platelet RNA enables accurate detection of ovarian cancer: an intercontinental, biomarker identification study

Platelets are reprogrammed by cancer via a process called education, which favors cancer development. The transcriptional profile of tumor-educated platelets (TEPs) is skewed and therefore practicable for cancer detection. This intercontinental, hospital-based, diagnostic study included 761 treatment-naïve inpatients with histologically confirmed adnexal masses and 167 healthy controls from nine medical centers (China, n = 3; Netherlands, n = 5; Poland, n = 1) between September 2016 and May 2019. The main outcomes were the performance of TEPs and their combination with CA125 in two Chinese (VC1 and VC2) and the European (VC3) validation cohorts collectively and independently. Exploratory outcome was the value of TEPs in public pan-cancer platelet transcriptome datasets. The AUCs for TEPs in the combined validation cohort, VC1, VC2, and VC3 were 0.918 (95% CI 0.889-0.948), 0.923 (0.855-0.990), 0.918 (0.872-0.963), and 0.887 (0.813-0.960), respectively. Combination of TEPs and CA125 demonstrated an AUC of 0.922 (0.889-0.955) in the combined validation cohort; 0.955 (0.912-0.997) in VC1; 0.939 (0.901-0.977) in VC2; 0.917 (0.824-1.000) in VC3. For subgroup analysis, TEPs exhibited an AUC of 0.858, 0.859, and 0.920 to detect early-stage, borderline, non-epithelial diseases and 0.899 to discriminate ovarian cancer from endometriosis. TEPs had robustness, compatibility, and universality for preoperative diagnosis of ovarian cancer since it withstood validations in populations of different ethnicities, heterogeneous histological subtypes, and early-stage ovarian cancer. However, these observations warrant prospective validations in a larger population before clinical utilities.

Cangxi Tongbi Capsules promote chondrocyte autophagy by regulating circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit development of knee osteoarthritis / 中国中药杂志

To investigate the mechanism by which Cangxi Tongbi Capsules promote chondrocyte autophagy to inhibit knee osteoarthritis(KOA) progression by regulating the circRNA＿0008365/miR-1271/p38 mitogen-activated protein kinase(MAPK) pathway. The cell and animal models of KOA were established and intervened with Cangxi Tongbi Capsules, si-circRNA＿0008365, si-NC, and Cangxi Tongbi Capsules combined with si-circRNA＿0008365. Flow cytometry and transmission electron microscopy were employed to determine the level of apoptosis and observe autophagosomes, respectively. Western blot was employed to reveal the changes in the protein levels of microtubule-associated protein light chain 3(LC3)Ⅱ/Ⅰ, Beclin-1, selective autophagy junction protein p62/sequestosome 1, collagen Ⅱ, a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS-5), and p38 MAPK. The mRNA levels of circRNA＿0008365, miR-1271, collagen Ⅱ, and ADAMTS-5 were determined by qRT-PCR. Hematoxylin-eosin staining was employed to reveal the pathological changes of the cartilage tissue of the knee, and enzyme-linked immunosorbent assay to measure the levels of interleukin-1β(IL-1β) and tumor necrosis factor-alpha(TNF-α). The chondrocytes treated with IL-1β showed down-regulated expression of circRNA＿0008365, up-regulated expression of miR-1271 and p38 MAPK, lowered autophagy level, increased apoptosis rate, and accelerated catabolism of extracellular matrix. The intervention with Cangxi Tongbi Capsules up-regulated the expression of circRNA＿0008365, down-regulated the expression of miR-1271 and p38 MAPK, increased the autophagy level, decreased the apoptosis rate, and weakened the catabolism of extracellular matrix. However, the effect of Cangxi Tongbi Capsules was suppressed after interfering with circRNA＿0008365. The in vivo experiments showed that Cangxi Tongbi Capsules dose-dependently inhibited the p38 MAPK pathway, enhanced chondrocyte autophagy, and mitigated articular cartilage damage and inflammatory response, thereby inhibiting the progression of KOA in rats. This study indicated that Cangxi Tongbi Capsules promoted chondrocyte autophagy by regulating the circRNA＿0008365/miR-1271/p38 MAPK pathway to inhibit the development of KOA.

Time-dependent injury of mouse cerebral cortex and hippocampus by acute hypoxia / 生理学报

The aim of this study was to investigate the harmful effects of acute hypoxia on mouse cerebral cortex and hippocampus and the underlying mechanism. Mouse model of acute hypoxia was constructed by using a sealed glass jar. Laser speckle contrast imaging was used to detect the changes of cerebral blood flow after different time duration of hypoxia. Total superoxide dismutase (T-SOD) and malondialdehyde (MDA) assay kits were used to detect oxidative stress in cerebral cortex and hippocampus. Immunofluorescent staining was used to detect neuroinflammatory response of microglia in the cerebral cortex and hippocampus. One-step TUNEL method was used to detect neuronal apoptosis. The results showed that, compared with non-hypoxia (0 min hypoxia) group, 30 min hypoxia group exhibited decreased cerebral blood flow, higher percentage of CD68+/Iba1+ microglia, and increased neural apoptosis in the cerebral cortex and hippocampus. Compared with 30 min group, 60 min hypoxia group showed significantly decreased cerebral blood flow, increased MDA content in the cortex, as well as greater percentage of CD68+/Iba1+ microglia and neuronal apoptosis in the cerebral cortex and hippocampus. These results suggest that acute hypoxia damages brain tissue in a time-dependent manner and the oxidative stress and neuroinflammation are important mechanisms.

A prospective randomized controlled study on the effects of bicycle ergometer rehabilitation training on quadriceps and walking ability of patients with lower limb dysfunction caused by extensive burns / 中华烧伤杂志

Objective: To explore the effects of bicycle ergometer rehabilitation training on quadriceps and walking ability of patients with lower limb dysfunction caused by extensive burns. Methods: A prospective randomized controlled study was conducted. A total of 40 patients with extensive burns who met the inclusion criteria and were admitted to Tongren Hospital of Wuhan University＆Wuhan Third Hospital from December 2017 to December 2020 were selected. According to the random number table, the patients were divided into conventional training group (16 males, 4 females, aged (45±10) years) and combined training group (13 males, 7 females, aged (39±8) years). Patients in conventional training group were given conventional rehabilitation therapy such as joint loosening, lower limb strength training, walking training, and pressure therapy, while patients in combined training group were given additional bicycle ergometer rehabilitation training on the basis of conventional rehabilitation. For patients in the 2 groups before and after a 2-month's treatment, the thickness of quadriceps was measured by ultrasonic diagnostic instrument, the muscle strength of quadriceps was measured by portable muscle strength tester, the walking ability was tested with a 6-min and a 10-meter walk tests, and the patients' satisfaction for treatment effects was assessed using the modified Likert scale. Data were statistically analyzed with independent or paired sample t test, Mann-Whitney U test, Wilcoxon signed rank test, or chi-square test. Results: After 2-month's treatment, the quadriceps thickness of patients in combined training group was (3.76±0.39) cm, which was significantly thicker than (3.45±0.35) cm in conventional training group (t=2.67, P<0.05); quadriceps thickness of patients in conventional training group and combined training group after 2-month's treatment was significantly thicker than that before treatment (with t values of 5.99 and 8.62, respectively, P<0.01). After 2-month's treatment, the quadriceps muscle strength of patients in combined training group was significantly greater than that in conventional training group (Z=2.69, P<0.01); quadriceps muscle strength of patients in conventional training group and combined training group after 2-month's treatment was significantly greater than that before treatment (with Z values of 3.92 and 3.92, respectively, P<0.01). After 2-month's treatment, the 6-min walking distance of patients in combined training group was (488±39) m, which was significantly longer than (429±25) m in conventional training group (t=5.66, P<0.01); the 6-min walking distance of patients after 2-month's treatment in conventional training group and combined training group was significantly longer than that before treatment (with t values of 13.16 and 17.92, respectively, P<0.01). After 2-month's treatment, the 10-meter walking time of patients in combined training group was significantly shorter than that in conventional training group (t=3.20, P<0.01); and the 10-meter walking time in conventional training group and combined training group was significantly shorter than that before treatment (with t values of 7.21 and 13.13, respectively, P<0.01). The patients' satisfaction score for treatment effects in combined training group was significantly higher than that in conventional training group (Z=3.14, P<0.01), and the patients' satisfaction scores for treatment effects in conventional training group and combined training group after 2-month's treatment were significantly greater than those before treatment (with Z values of 3.98 and 4.04, respectively, P<0.01). Conclusions: Bicycle ergometer rehabilitation training can be used to improve quadriceps thickness, muscle strength, and walking ability of patients with lower limb dysfunction caused by extensive burns. It can also improve the satisfaction of patients with the treatment outcome, and therefore is worthy of promotion.

Retrospective analysis of 35 burn patients in different stages of pregnancy / 中华烧伤杂志

Objective: To summarize the clinical outcomes of burn patients in different stages of pregnancy and explore a rational therapeutic scheme for burns during pregnancy. Methods: A retrospective observational study was conducted. From June 2010 to June 2020, 21 patients who met the inclusion criteria were admitted to the Department of Burns of Wuhan Third Hospital and 14 patients who met the inclusion criteria were admitted to the Department of Burns of the First Affiliated Hospital of Nanchang University. Based on the pregnancy period when patients suffered burns, the 35 patients were divided into early pregnancy group with 18 patients (aged (26±4) years, with 8 (4, 11) weeks of gestation), middle pregnancy group with 10 patients (aged (26±3) years, with 21 (14, 27) weeks of gestation), and late pregnancy group with 7 patients (aged (30±5) years, with 32 (29, 35) weeks of gestation). All the patients received treatment including fluid resuscitation, anti-infection, wound treatment, and multidisciplinary comprehensive managements. The burn-related complications during the treatment, maternal outcomes, fetal outcomes, fetal delivery mode, gestational weeks at delivery, and newborn weight of patients in the 3 groups were recorded. Data were statistically analyzed with one-way analysis of variance, Kruskal-Wallis test, and Fisher's exact probability test. Results: During the treatment, there were 4, 4, and 2 patients who suffered wound infections and 1, 3, and 2 patients who developed shock symptoms, respectively, in early pregnancy group, middle pregnancy group, and late pregnancy group. There were no statistically significant differences in them among the 3 groups (P>0.05). One patient in late pregnancy group developed into multiple organ dysfunction syndrome after debridement. At last, all the pregnant women survived, and no statistically significant difference existed among the 3 groups (P>0.05). In early pregnancy group, middle pregnancy group, and late pregnancy group, the survived fetus cases were 9, 8, and 6, respectively, and the differences between them were not statistically significant (P>0.05). Variables including stillbirth and full-term birth were close in patients in the 3 groups (P>0.05), while the preterm birth and miscarriage in patients in the 3 groups were statistically different (P<0.05 or P<0.01), with the early pregnancy group having the most miscarriage cases and the fewest preterm birth cases. There were no statistically significant differences in fetal delivery mode, gestational weeks at delivery, and newborn weight among the patients with survived fetus in 3 groups (P>0.05). Conclusions: For patients suffering burns during early, middle, and late pregnancy, superior rates of maternal and fetal survival can be achieved after timely and adequate treatments including fluid resuscitation, anti-infection, wound treatment, and multidisciplinary comprehensive managements.

Clinical effects of free transplantation of expanded ilioinguinal flaps in the reconstruction of severe scar contracture deformity after extensive burns / 中华烧伤杂志

Objective: To investigate the clinical effects of free transplantation of expanded ilioinguinal flaps in the reconstruction of severe scar contracture after extensive burns. Methods: A retrospective observational study was conducted. From August 2017 to October 2021, 7 patients with severe scar contracture deformity caused by extensive burns were hospitalized in Tongren Hospital of Wuhan University & Wuhan Third Hospital, including 5 males and 2 females, aged 26-65 years, with scar area of 20 cm×4 cm-34 cm×14 cm. In the first stage, the rectangular skin and soft tissue expander (hereinafter referred to as the expander) with rated capacity of 500-600 mL were embedded above the inguinal ligament, and then normal saline was injected after stitch removal for expansion to meet the needs of repair surgery. In the second stage, the scar was removed by surgical excision to correct the deformity and release the adhesion and contracture; after the removal of the expanders, the expanded ilioinguinal free flaps were harvested. When a larger flap was needed, the paraumbilical perforator flap was harvested at the same time, and the flaps were transplanted to the secondary wound after scar resection. The number of embedded expanders, the total amount of injected normal saline, the expansion time, the complications of skin and soft tissue expansion, the number, area, thickness, and anastomotic vascular pedicles of the expanded ilioinguinal flaps being resected, the type of flaps used, the repair method of flap donor sites, and the survival of flaps after operation were observed and recorded. The long-term repair effect and donor site condition were followed up. At the last follow-up, the patients' satisfaction with the curative effect of each surgical site was investigated according to the grade 5 score of Likert scale. Results: A total of 10 expanders were embedded in 7 patients, of which 4 patients had 1 each and 3 patients had 2 each. The total volume of normal saline injected was 800-1 800 (1 342±385) mL, and the expansion time was 4-24 (11±5) months. One patient had the expander exposed due to infection after the expander being inserted, while the other patients had no complications of skin and soft tissue expansion. Totally 10 expanded ilioinguinal flaps with the area of 22 cm×6 cm-36 cm×16 cm ((326±132) cm2) and the thickness of 0.6-1.1 (0.77±0.16) cm were harvested. Among the 10 expanded ilioinguinal flaps, 5 were pedicled with the superficial circumflex iliac artery, 3 with the superficial abdominal artery with relatively large caliber, 1 with the common trunk of the superficial circumflex iliac artery and the superficial abdominal artery, and 1 flap was anastomosed with the superficial circumflex iliac artery and bridged the superficial abdominal artery for intra-arterial supercharge. Unilateral expanded ilioinguinal flap combined with ipsilateral paraumbilical perforator flap were harvested in 4 cases, bilateral expanded ilioinguinal flaps were harvested in 1 case, and unilateral expanded ilioinguinal flap was harvested in 2 cases. Except for 1 case being transplanted with autologous split-thickness scalp to repair the flap donor site after combined resection of bilateral expanded ilioinguinal flaps, the donor sites of the other patients were sutured directly. All the flaps survived after operation without tip necrosis or wound residue. Follow-up for 3-30 (15±10) months showed that the flap was soft and not bloated, the function and appearance of the recipient area were significantly improved compared with those before operation, and the appearance of the donor sites was good. At the last follow-up, the patients' satisfaction with the treatment effect of the surgical site scored 4-5 (4.5±0.4). Conclusions: The expanded ilioinguinal flap can be obtained in a large area. It has the advantages of rich blood supply, less damage to the donor site, concealed location, and being convenient to be resected and transplanted in combination with the paraumbilical perforator flap. It is suitable for the clinical reconstruction and treatment of severe scar contracture deformity after extensive burns.

Hypoxic pretreatment enhancing the angiogenesis of aged human bone marrow mesenchymal stem cells through hypoxia inducible factor-1α / 解剖学报

Objective To investigate the effect of hypoxic pretreatment on the angiogenesis of aged human bone marrow mesenchymal stem cells (hBMSCs), so to provide experimental support for more effective autologous stem cell transplantation therapy in aged patients with ischemic myocardial injury. Methods The aged hBMSCs were cultured in a hypoxic incubator, and then the cell morphology was observed under inverted phase-contrast microscope, the surface markers were detected by flow cytometry, and the differentiation potential was detected by osteogenic and adipogenic differentiation. Subsequently, the conditioned medium (CM) of young hBMSCs under normoxic culture (norCM), the conditioned media of aged hBMSCs under normoxic and hypoxic culture(hypoCM) were collected respectively. They were named as norCM-young, norCM-old and hypoCM-old. The equal volume of medium, which was not treated with stem cells, was set as control group. Human umbilical vein endothelial cells (HUVECs) were treated with 4 conditioned media, the cell survival rate was detected by CCK-8 assay, and the tube formation experiment was used to detect the angiogenesis ability in vitro. The BCA method was used to detect the total protein concentration of the conditioned medium of each group, and the Western blotting was used to detect the expression of hypoxia inducible factor-1α (HIF-1α) in the cells and the conditioned media. Results There was not significant effect of hypoxic pretreatment on the morphology, surface markers and differentiation ability of aged hBMSCs (P > 0. 05. n ≥ 3). Compared with the norCM-old group, hypoCM-old group significantly improved the survival of HUVECs under hypoxia-reoxygenation condition (P < 0. 05, n = 5), and the tube formation ability of it (P<0. 01, n = 5). The total protein concentration of hypoCM-old group was significantly higher than that of norCM-old group (P<0. 05, n = 3). The expression of HIF-1α in hBMSCs of hypo-old group was significantly higher than that of nor-old group (P<0. 05,n = 3), while the content of HIF-1α in conditioned medium of hypoCM-old group was significantly higher than that in norCM-old group (P<0. 01,n = 3). Conclusion The aged hBMSCs pretreated with hypoxia can promote the survival and tube formation of HUVECs through paracrine in vitro, which is HIF-1α-related.

Suitable soil moisture contents for water use efficiency and saponins accumulation in Panax notoginseng / 中草药·英文版

Objective: The moisture content in the soil directly affects the yield and quality of Panax notoginseng, especially at the age of three years old. However, the suitable moisture for the growth of P. notoginseng is unknown. In this study, the effects of different soil moisture on the growth of P. notoginseng were studied. Methods: Four different water treatments (0.45 field capacity (FC), 0.60 FC, 0.70 FC, and 0.85 FC) were set up in Shilin County, Yunnan Province, China. The water consumption and daily dynamic of water consumption were determined daily (from April 21 to October 18, 2012), and the daily dynamic of water consumption under different weather conditions (sunny and rainy) was determined. The transpiration coefficient and water use efficiency were calculated through dry matter accumulation and total water consumption. Accumulation of saponins of roots of P. notoginseng were analyzed by HPLC after treated, and the soil moisture content suitable for the growth of P. notoginseng was estimated by regression fitting of the active ingredient accumulation and the soil moisture content. Results: The water consumption of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.89, 3.68, 3.37 and 2.73 kg/plant per day, respectively. The water consumption of P. notoginseng from June to August was greater than other months. The daily dynamic of water consumption on sunny days and sunny days after rain showed a “double peak” feature, and it showed a “single peak” feature on rainy days. The water uses efficiency (WUE) of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.51, 3.32, 4.59, 3.39 gDW/kg H

Two naturally derived small molecules disrupt the sineoculis homeobox homolog 1-eyes absent homolog 1 (SIX1-EYA1) interaction to inhibit colorectal cancer cell growth / 中华医学杂志(英文版)

BACKGROUND@#Emerging evidence indicates that the sineoculis homeobox homolog 1-eyes absent homolog 1 (SIX1-EYA1) transcriptional complex significantly contributes to the pathogenesis of multiple cancers by mediating the expression of genes involved in different biological processes, such as cell-cycle progression and metastasis. However, the roles of the SIX1-EYA1 transcriptional complex and its targets in colorectal cancer (CRC) are still being investigated. This study aimed to investigate the roles of SIX1-EYA1 in the pathogenesis of CRC, to screen inhibitors disrupting the SIX1-EYA1 interaction and to evaluate the efficiency of small molecules in the inhibition of CRC cell growth.@*METHODS@#Real-time quantitative polymerase chain reaction and western blotting were performed to examine gene and protein levels in CRC cells and clinical tissues (collected from CRC patients who underwent surgery in the Department of Integrated Traditional and Western Medicine, West China Hospital of Sichuan University, between 2016 and 2018, n = 24). In vivo immunoprecipitation and in vitro pulldown assays were carried out to determine SIX1-EYA1 interaction. Cell proliferation, cell survival, and cell invasion were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, clonogenic assay, and Boyden chamber assay, respectively. The Amplified Luminescent Proximity Homogeneous Assay Screen (AlphaScreen) method was used to obtain small molecules that specifically disrupted SIX1-EYA1 interaction. CRC cells harboring different levels of SIX1/EYA1 were injected into nude mice to establish tumor xenografts, and small molecules were also injected into mice to evaluate their efficiency to inhibit tumor growth.@*RESULTS@#Both SIX1 and EYA1 were overexpressed in CRC cancerous tissues (for SIX1, 7.47 ± 3.54 vs.1.88 ± 0.35, t = 4.92, P = 0.008; for EYA1, 7.61 ± 2.03 vs. 2.22 ± 0.45, t = 6.73, P = 0.005). The SIX1/EYA1 complex could mediate the expression of two important genes including cyclin A1 (CCNA1) and transforming growth factor beta 1 (TGFB1) by binding to the myocyte enhancer factor 3 consensus. Knockdown of both SIX1 and EYA1 could decrease cell proliferation, cell invasion, tumor growth, and in vivo tumor growth (all P < 0.01). Two small molecules, NSC0191 and NSC0933, were obtained using AlphaScreen and they could significantly inhibit the SIX1-EYA1 interaction with a half-maximal inhibitory concentration (IC50) of 12.60 ± 1.15 μmol/L and 83.43 ± 7.24 μmol/L, respectively. Administration of these two compounds could significantly repress the expression of CCNA1 and TGFB1 and inhibit the growth of CRC cells in vitro and in vivo.@*CONCLUSIONS@#Overexpression of the SIX1/EYA1 complex transactivated the expression of CCNA1 and TGFB1, causing the pathogenesis of CRC. Pharmacological inhibition of the SIX1-EYA1 interaction with NSC0191 and NSC0933 significantly inhibited CRC cell growth by affecting cell-cycle progression and metastasis.

Intervention Mechanism of Yishen Huayu Prescription on Glomerular Podocyte Injury in Diabetic Nephropathy Rats Based on EMT Regulated by Wnt/β-catenin Pathway / 中国实验方剂学杂志

Objective:To investigate the intervention mechanism of Yishen Huayu prescription on glomerular podocyte injury in diabetic nephropathy （DN） rats based on epithelialmesenchymal transition （EMT） regulated by Wnt/<italic>β</italic>-catenin pathway. Method:The 60 SD rats were divided into control group， model group， Wnt-C59 group （0.03 g·kg^-1 Wnt/<italic>β</italic>-catenin pathway inhibitor）， low-dose group （8 g·kg^-1）， medium-dose group （16 g·kg^-1） and high-dose group （32 g·kg^-1）. After 12 weeks， various indexes ， including general signs， serum creatinine （SCr）， blood urea nitrogen （BUN）， renal index， urinary protein， blood glucose， renal pathological changes， podocyte and expressions of glomerular basement membrane injury and podocyte injury related proteins ［nephrin， synaptopodin］， Wnt/<italic>β</italic>-catenin pathway related proteins （Wnt1， <italic>β</italic>-catenin）， podocyte EMT related protein ［<italic>α</italic>-smooth muscle actin （<italic>α</italic>-SMA）， E-cadherin］， were compared between groups. Result:Compared with the control group， the renal tissue in the model group showed significant pathological changes， including diffuse thickening of glomerular mesangial matrix and severe foot process fusion， and a significant increase in SCr， BUN， renal indexes， urinary protein， blood glucose， Wnt1， <italic>β</italic>-catenin， and <italic>α</italic>-SMA expression levels （<italic>P</italic><0.05） as well as a significant decrease in nephrin， synaptopodin and E-cadherin expression levels（<italic>P</italic><0.05）. Compared with model group， SCr， BUN， renal index， urinary protein， blood glucose， Wnt1， <italic>β</italic>-catenin， and <italic>α</italic>-SMA expression levels in each intervention group significantly decreased （<italic>P</italic><0.05）， while the expression levels of nephrin， synaptopodin and E-cadherin significantly increased （<italic>P</italic><0.05）. Among intervention groups， the improvement of above indexes in high-dose Yishen Huayu prescription group was the most obvious （<italic>P</italic><0.05）， which was similar to the effect in Wnt-C59 group. Conclusion:Yishen Huayu prescription prevents podocyte EMT by inhibiting Wnt/<italic>β</italic>-catenin pathway， thereby repairing glomerular podocyte injury in rats with diabetic nephropathy.

Prognostic Factors Analysis of Children with Hemophagocytic Lymphohistiocytosis / 中国实验血液学杂志

OBJECTIVE@#To analyze the risk factors affecting prognosis of children with hemophagocytic lymphohistiocytosis (HLH).@*METHODS@#The clinical manifestations and laboratory data of 143 HLH children who met the HLH-2004 diagnostic criteria in Shenzhen Children's Hospital from January 2009 to May 2017 were retrospectively analyzed, and the independent factors affecting prognosis were also analyzed.@*RESULTS@#The median age of 143 HLH children was 1.9 (0.1-14.3) years old, and the median follow-up time was 6.7 years (1 day - 11.9 years). The overall survival rate of 1 month, 1 year, and 10 years was (87.4±5.5)%, (81.1±6.5)%, and (81.1±6.5)%, respectively. The deaths occurred within 1 year after onset. Multivariate analysis showed that central nervous system (CNS) involvement (P=0.047), low hemoglobin (P=0.002), prolonged activated partial thromboplastin time (APTT) (P<0.001), high triglyceride (P=0.005) were all the independent risk factors affecting survival of the children. Receiver operating characteristic curve indicated that APTT (AUC=0.753, P<0.001) was more valuable than other risk factors in predicting death of the children. The cut-off value of APTT was 56.6 s, and the sensitivity and specificity of which was 55.6% and 89.7%, respectively.@*CONCLUSION@#Hypohemoglobinemia, prolonged APTT, hypertriglyceridemia, and CNS involvement the risk factors affecting prognosis of HLH, and prolonged APTT shows a strong predictive value for death.

Safety and effectiveness of oxaliplatin combined with capecitabine or oxaliplatin combined with S-1 neoadjuvant chemotherapy in the treatment of advanced gastric cancer / 中华胃肠外科杂志

Objective: To explore the safety and efficacy of oxaliplatin plus capecitabine (CapeOX) or oxaliplatin plus S-1 (SOX) regimen neoadjuvant chemotherapy in the treatment of advanced gastric cancer. Methods: A retrospective cohort study was performed. Clinical data of patients diagnosed as advanced gastric cancer undergoing CapeOX/SOX neoadjuvant chemotherapy and standard laparoscopic radical operation for gastric cancer in Ruijin Hospital of Shanghai Jiaotong University School of Medicine from April 2016 to April 2019 were retrospectively collected. Inclusion criteria were as follows: (1) age≥18 years; (2) gastric adenocarcinoma was confirmed by histopathology and the clinical stage was T3-4aN+M0; (3) tumor could be resectable; (4) preoperative neoadjuvant chemotherapy was CapeOX or SOX regimen without radiotherapy or other regimen chemotherapy; (5) no other concurrent malignant tumor; (6) the Eastern Cooperative Oncology Group (ECOG) score ≤ 1; (7) no bone marrow suppression; (8) normal liver and kidney function. Exclusion criteria were as follows: (1) patients with recurrent gastric cancer; (2) patients receiving emergency surgery due to tumor perforation, bleeding, obstruction, etc.; (3) allergy to oxaliplatin, S-1, capecitabine or any drug excipients; (4) diagnosed with coronary heart disease, cardiomyopathy, or the New York Heart Association class III or IV; (5) pregnant or lactating women. A total of 118 patients were enrolled as the neoadjuvant chemotherapy group, and 379 patients with locally advanced gastric cancer who received surgery combined with postoperative adjuvant chemotherapy over the same period simultaneously were included as the adjuvant chemotherapy group. After propensity score matching was performed including gender, age, ECOG score, tumor site, clinical stage, chemotherapy regimen and other factors by 1:1 ratio, there were 40 cases in each group. The differences between the two groups in general conditions, efficacy of neoadjuvant chemotherapy, intraoperative conditions, postoperative conditions, histopathological results, chemotherapy-related adverse events, and survival status were compared and analyzed. Results: Comparison of baseline demographics between the two groups showed no statistically significant difference (all P>0.05). In the neoadjuvant chemotherapy group, 5.0% (2/40) of patients achieved clinical complete response, 57.5% (23/40) achieved partial response, 32.5% (13/40) remained stable disease, and 5.0% (2/40) had disease progression before surgery. Objective response rate was 62.5% (25/40), and disease control rate was 95.0% (38/40). There were no statistically significant differences between neoadjuvant chemotherapy group and adjuvant chemotherapy group in terms of operation time, intraoperative blood loss, number of lymph node harvested, length of postoperative hospital stay, and postoperative mortality and morbidity (all P>0.05). Postoperative complications were well managed with conservative treatment. No Clavien-Dindo IV or V complications were observed in both groups. Pathological results showed that the proportion of patients with pathological stage T1 in the neoadjuvant chemotherapy group was significantly higher than that in the adjuvant chemotherapy group [27.5% (11/40) vs. 5.0% (2/40)], while the proportion of patients with pathological stage T3 was significantly lower than that in the adjuvant chemotherapy group [20.0% (8/40) vs. 45.0% (18/40)], with statistically significant difference (χ(2)=15.432, P=0.001). In the neoadjuvant chemotherapy group, there were 4 cases of tumor regression grade 0, 8 cases of grade 1, 16 cases of grade 2, and 12 cases of grade 3. The pathological complete response rate was 10% (4/40), the overall pathological response rate was 70.0% (28/40). There was no statistically significant difference in the incidence of chemotherapy-related adverse events between neoadjuvant chemotherapy group and adjuvant chemotherapy group [40% (16/40) vs. 37.5% (15/40), P>0.05). There were no statistically significant differences in OS (43 months vs. 40 months) and 3-year OS rate (66.1% vs. 59.8%) between neoadjuvant chemotherapy group and adjuvant chemotherapy group (P=0.428). The disease-free survival (DFS) and 3-year DFS rates of the neoadjuvant chemotherapy group were significantly superior to those of the adjuvant chemotherapy group (36 months vs. 28 months, 51.4% vs. 35.8%, P=0.048). Conclusion: CapeOX or SOX regimen neoadjuvant chemotherapy is a safe, effective and feasible treatment mode for advanced gastric cancer without increasing surgical risk and can improve the DFS of patients.

Effect of early intervention of electroacupuncture on learning-memory ability and level of hippocampal phosphorylated Tau protein in SAMP8 mice / 中国针灸

OBJECTIVE@#To explore the effect of early intervention electroacupuncture (EA) at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) on the learning-memory ability and the expression of phosphorylated Tau protein in the hippocampus of SAMP8 mice, so as to provide reference for the intervening period of EA for Alzheimer's disease (AD).@*METHODS@#A total of 36 3-month old SAMP8 mice were randomly divided into a model group, a 3-month-old EA group and a 9-month-old EA group, 12 mice in each group. Twelve normal SAMR1 mice with the same age were taken as the control group. The mice in the 3-month-old EA group and 9-month-old EA group were treated with EA at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) separately 3 months old and 9 months old (continuous wave, 2 Hz, 1.5-2 mA), 20 min each time, once a day, 8 days as a course of treatment, with an interval of 2 days between courses, totally 3 courses of treatment were given. The mice sample in each group was collected at the age of 10 months after the learning-memory ability tested by Morris water maze. The expression of phosphorylated Tau protein in the hippocampus was detected by immunohistochemistry and Western blot, and the expression of Tau mRNA was detected by real-time PCR.@*RESULTS@#Compared with the control group, in the model group, the escape latency was significantly increased (<0.01), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were reduced (<0.01), and the expressions of phosphorylated Tau protein and Tau mRNA in hippocampus were increased (<0.01). Compared with the model group, in the 3-month-old EA group and 9-month-old EA group, the escape latency was significantly reduced (<0.05), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were increased (<0.05), and the expressions of phosphorylated Tau protein and Tau mRNA in hippocampus were reduced (<0.05). Compared with the 9-month-old EA group, in the 3-month-old EA group, the escape latency was significantly reduced (<0.05), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were increased (<0.05), and the expressions of phosphorylated Tau protein and Tau mRNA were reduced (<0.01).@*CONCLUSION@#The early EA intervention could more effectively improve the learning-memory ability and inhibit phosphorylation of Tau protein in the hippocampus of SAMP8 mice.

Effects of Virtual Reality Balance Training on Proprioception of Knee after Anterior Cruciate Ligament Reconstruction / 中国康复理论与实践

Objective:To investigate the effect of virtual reality balance training on knee proprioception after anterior cruciate ligament reconstruction (ACLR). Methods:From January, 2016 to December, 2018, 72 patients after ACLR were randomly divided into control group (n = 36) and observation group (n = 36). On the basis of routine rehabilitation, they received routine balance training and balance training with virtual reality, respectively, for eight weeks. Before and after training, the center of pressure (COP) time stability, the COP speed (COPS) of closed eyes, the COPS of one leg, the knee joint reaction time, the center of pressure area (COPA) and the active joint angle recurrence deviation were measured. Results:After training, the COP time stability, the COPS of closed eyes, the COPS of one leg, knee joint reaction time, COPA and active joint angle recurrence deviation improved in both groups (|t|>5.611, P < 0.001); and the COP time stability, the COPS of closed eyes, the COPS of one leg, the knee joint reaction time and active joint angle recurrence deviation were better in the observation group than in the control group (|t| > 3.232, P < 0.01), however, no significant difference was found in COPA between two groups (t = 1.054, P = 0.296). Conclusion:Virtual reality balance training can improve the proprioception of knee joint after ACLR, which is better than routine balance training.

Antibacterial activity of bacillomycin D-like compounds isolated from Bacillus amyloliquefaciens HAB-2 against Burkholderia pseudomallei

Objective: To investigate the inhibitory effect on Burkholderia pseudomallei (B. pseudomallei) strain HNBP001 of a bacillomycin D-like cyclic lipopeptide compound named bacillomycin DC isolated from Bacillus amyloliquefaciens HAB-2. Methods: The antibacterial effect of bacillomycin DC on B. pseudomallei was determined using the disk diffusion method. The minimum inhibitory concentrations were evaluated by microdilution assay. In addition, transmission electron microscopy was performed and quantitative real-time polymerase chain reaction assay was carried out to determine the expression of MexB, OprD2, and qnrS genes. Results: Bacillomycin DC produced an inhibition zone against B. pseudomallei with minimum inhibitory concentration values of 12.5 μg/mL 24 h after treatment and 50 μg/mL at 48 and 72 h. Transmission electron microscopy showed that bacillomycin DC resulted in roughening cell surface and cell membrane damage. Quantitative real-time polymerase chain reaction analysis showed low expression of MexB, OprD2 and qnrS genes. Conclusions: Bacillomycin DC inhibits the growth of B. pseudomallei and can be a new candidate for antimicrobial agents of B. pseudomallei.

Mechanism of salicylic acid ameliorates salt-induced changes in Andrographis paniculata / 中国中药杂志

In this study, Andrographis paniculata seedlings were used as experimental materials to study the effects of salicylic acid(SA) on the growth and effective component accumulation of A. paniculata under NaCl stress. The results showed that with the increase of NaCl concentration, the growth of A. paniculata seedlings was significantly inhibited, and the content of carotene and carotenoid decreased. The activity of antioxidant enzyme was enhanced. At the same time, the contents of proline, proline and soluble protein were on the rise. The contents of andrographolide, new andrographolide and deoxyandrographolide showed an upward trend, while deoxyandrographolide showed a downward trend. Treatment with 100 mmol·L~(-1) NaCl+5 mg·L~(-1) SA showed a significant increase in antioxidant enzyme activity in A. paniculata leaves. Treatment with 100 mmol·L~(-1) NaCl+10 mg·L~(-1) SA showed significant changes in soluble protein and proline content in A. paniculata leaves, while MDA content in A. paniculata leaves significantly decreased. 10 mg·L~(-1) SA had the best effect on the growth of A. paniculata seedlings under salt stress. Under the treatment of 50 mmol·L~(-1) NaCl+10 mg·L~(-1) SA, fresh weight, dry weight and leaf dry weight of A. paniculata seedlings reached the highest level, which were 1.02, 1.09 and 1.11 times of those in the control group, respectively. The concentrations of NaCl and 10 mg·L~(-1) SA were significantly higher than those of the control group. Four key enzyme genes of A. paniculata diterpene lactone synthesis pathway were selected to explore the molecular mechanism of salicylic acid to alleviate salt stress. With the increase of salt stress, the relative expressions of HMGR, GGPS and ApCPS were up-regulated, indicating that salt stress may enhance the synthesis of A. paniculata diterpene lactone through MVA pathway. SA can effectively promote the growth and development of A. paniculata under salt stress, improve its osmotic regulation and antioxidant capacity, improve its salt tolerance, and alleviate the effects of salt stress on A. paniculata.

Research progress of Rehmanniae Radix in prevention and treatment of osteoporosis / 中国中药杂志

Osteoporosis fracture with high disability and mortality is a difficult problem that seriously affects the life quality of individuals. At present, there is still a lack of anti-osteoporosis drugs with clear target and significant efficacy in the clinical practice. Rehmanniae Radix and its prescriptions have significant clinical effects. In this regard, more and more studies have reported the effects and mechanisms of Rehmanniae Radix and its active components, and the certain research outputs have been achieved. In this article, the PubMed, Web of science, China National Knowledge Infrastructure, and Wanfang database were searched to collect and organize the latest research progress of Rehmanniae Radix treatment of osteoporosis in the recent 10 years. We summarized the research dynamics as well as the function indexes and mechanisms of the raw and processed Rehmanniae Radix, active ingredients such as catalpol, aucubin, acteoside and Rehmanniae Radix polysaccharide, and their formulating prescriptions, and then excavated the potential active ingredients, targets and signaling pathways, including the effect on bone marrow mesenchymal stem cells, promoting the osteoblast proliferation and promoting osteogenesis differentiation(increasing alkaline phosphatase, typeⅠ collagen, osteoprotegerin, and osteocalcin and promoting calcium deposits), increasing the bone density, inhibiting the osteoclast quantity and differentiation, promoting the osteoclast apoptosis, and reducing tartrate resistant acid phosphatase and bone resorption pit area to provide the reference and develop new ideas for developing Rehmanniae Radix prescriptions for treatment of osteoporosis and exploring its mechanism.

Transforaminal percutaneous endoscopic lumbar discectomy combined with anchorage technique of pedicle for the treatment of high prolapse free lumbar disc herniation / 中国骨伤

OBJECTIVE@#To investigate the clinical effects of percutaneous spinal endoscopy (percutaneous endoscopy) in the treatment of high prolapse free lumbar disc herniation.@*METHODS@#From May 2016 to June 2018, 24 patients with highly prolapse free lumbar disc herniation were enrolled in this study, including 11 males and 13 females, ranging in age from 48 to 72 years old, with an average of (59.5±7.2) years old. There were 1 case of L, 5 cases of L, 18 cases of L. The course of disease ranged from 8 to 26 months, with an average of (16.2±6.3) months. All the patients were subjected to local infiltration anesthesia. The clinical outcomes were evaluated by visual analog scale (VAS) for leg pain, low backpain and Oswestry Disability Index (ODI) at preoperative, first day after operation and 6 month after surgery. All data were statisticed by SPSS 22.0.@*RESULTS@#All the patients were followed up, and the duration ranged from 12 to 24 months, with a mean of (17.5±5.3) months. The average operation time was(69.8±14.2) minutes. One patient had cerebrospinal fluid leakage, which improved after supine rest. VAS scores of lower back pain were 6.36±1.27, 3.94±1.03, 1.62±0.87, 0.44±0.27, 0.37±0.29. VAS scores of leg pain were 8.28±1.74, 3.16±1.24, 2.83±1.13, 0.83±0.31, 0.46±0.31, and the differences were statistically significant (<0.05). The ODI were (48.79±9.83)%, (36.51±11.24)%, (21.05±6.35)%, (9.83±4.62)% and (7.24±4.72)% 1 day before and 1 weeks, 3 months, 6 months and 1 year after the operation, respectively. One year after the operation, the modified Macnab evaluation system was used to evaluate the clinical efficacy of the patients, 19 patients got an excellent result, 3 good, 2 fair and 0 poor.@*CONCLUSION@#Percutaneous endoscopic pedicle anchoring technique for the treatment of high prolapse free lumbar disc herniation can effectively improve the clinical symptoms of patients, and has the advantages of less trauma, less bleeding, rapid recovery, complete removal of the nucleus pulposus, and less pain of patients, etc., its clinical efficacy is accurate, operability is strong, it is worth promoting in the clinical use.