Stigmasterol protects human brain microvessel endothelial cells against ischemia-reperfusion injury through suppressing EPHA2 phosphorylation / 中国天然药物

Stigmasterol is a plant sterol with anti-apoptotic, anti-oxidative and anti-inflammatory effect through multiple mechanisms. In this study, we further assessed whether it exerts protective effect on human brain microvessel endothelial cells (HBMECs) against ischemia-reperfusion injury and explored the underlying mechanisms. HBMECs were used to establish an in vitro oxygen and glucose deprivation/reperfusion (OGD/R) model, while a middle cerebral artery occlusion (MCAO) model of rats were constructed. The interaction between stigmasterol and EPHA2 was detected by surface plasmon resonance (SPR) and cellular thermal shift assay (CETSA). The results showed that 10 μmol·L-1 stigmasterol significantly protected cell viability, alleviated the loss of tight junction proteins and attenuated the blood-brain barrier (BBB) damage induced by OGD/R in thein vitro model. Subsequent molecular docking showed that stigmasterol might interact with EPHA2 at multiple sites, including T692, a critical gatekeep residue of this receptor. Exogenous ephrin-A1 (an EPHA2 ligand) exacerbated OGD/R-induced EPHA2 phosphorylation at S897, facilitated ZO-1/claudin-5 loss, and promoted BBB leakage in vitro, which were significantly attenuated after stigmasterol treatment. The rat MCAO model confirmed these protective effects in vivo. In summary, these findings suggest that stigmasterol protects HBMECs against ischemia-reperfusion injury by maintaining cell viability, reducing the loss of tight junction proteins, and attenuating the BBB damage. These protective effects are at least meditated by its interaction with EPHA2 and inhibitory effect on EPHA2 phosphorylation.

Pharmacodynamic mechanism study on Modified sanhuang ointment in the treatment of anal ulcers and swelling in rats / 中国药房

OBJECTIVE To explore the improvement mechanism of Modified sanhuang ointment on anal ulcers and swelling model rats. METHODS The anal ulcer model of the rat was induced by using glacial acetic acid； the effects of Modified sanhuang ointment low-dose， medium-dose and high-dose groups （185， 370， 740 mg/kg）， western medicine positive control group （Compound carraghenates cream， 1 g/kg） and TCM positive control group （Mayinglong shexiang zhichuang ointments， 1 g/kg） on body weight， area of anal ulcer， grade of anal ulcer were investigated. The other groups of rats were used to induce rectal swelling models with croton oil； the effects of Modified sanhuang ointment low-dose， medium-dose and high-dose groups （185， 370， 740 mg/kg）， western medicine positive control group （Compound carraghenates cream， 1 g/kg） and TCM positive control group （Mayinglong shexiang zhichuang ointments， 1 g/kg） on the rate of rectal and anal swelling， serum contents of inflammatory factors [interleukin 2 （IL-2）， IL-4， tumor necrosis factor （TNF-α）]， pathological morphology of rectal tissue， the expression of transient receptor potential channel V1 （TRPV1） and substance P in rectal tissue and rectal vascular permeability were investigated. RESULTS In Modified sanhuang ointment， the increase in body weight was enhanced， and the area of anal ulcers， as well as the grade of anal ulcers in rats with anal ulcer models， were reduced to varying degrees； rectal tissue damage in rectal swelling model rats was improved； the rate of rectal and anal swelling， the serum contents of inflammatory factors， the expressions of TRPV1 and substance P in rectal tissue， and rectal vascular permeability were all decreased （P＜0.05 or P＜0.01）. The effect of Modified sanhuang ointment was better than that of western medicine positive control and TCM positive control.Modified sanhuang ointment can improve anal ulcers and swelling in rats by reducing the release of inflammatory factors， inhibiting the expression of TRPV1 and substance P.

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

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

Effects of in vivo targeted carboxylesterase 1f gene knockdown on the Kupffer cells polarization activity in mice with acute liver failure / 中华肝脏病杂志

Objective: To investigate the effect of targeted carboxylesterase 1f (Ces1f) gene knockdown on the polarization activity of Kupffer cells (KC) induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN) in mice with acute liver failure. Methods: The complex siRNA-EndoPorter formed by combining the small RNA (siRNA) carrying the Ces1f-targeting interference sequence and the polypeptide transport carrier (Endoporter) was wrapped in β-1, 3-D glucan shell to form complex particles (GeRPs). Thirty male C57BL/6 mice were randomly divided into a normal control group, a model group (LPS/D-GalN), a pretreatment group (GeRPs), a pretreatment model group (GeRPs+LPS/D-GalN), and an empty vector group (EndoPorter). Real-time fluorescent quantitative PCR and western blot were used to detect Ces1f mRNA and protein expression levels in the liver tissues of each mouse group. Real-time PCR was used to detect the expression levels of KC M1 polarization phenotypic differentiation cluster 86(CD86) mRNA and KC M2 polarization phenotypic differentiation cluster 163 (CD163) mRNA in each group. Immunofluorescence double staining technique was used to detect the expression of Ces1f protein and M1/M2 polarization phenotype CD86/CD163 protein in KC. Hematoxylin-eosin staining was used to observe the pathological damage to liver tissue. A one-way analysis of variance was used to compare the means among multiple groups, or an independent sample nonparametric rank sum test was used when the variances were uneven. Results: The relative expression levels of Ces1f mRNA/protein in liver tissue of the normal control group, model group, pretreatment group, and pretreatment model group were 1.00 ± 0.00, 0.80 ± 0.03/0.80 ± 0.14, 0.56 ± 0.08/0.52 ± 0.13, and 0.26 ± 0.05/0.29 ± 0.13, respectively, and the differences among the groups were statistically significant (F = 9.171/3.957, 20.740/9.315, 34.530/13.830, P < 0.01). The percentages of Ces1f-positive Kupffer cells in the normal control group, model group, pretreatment group, and pretreatment model group were 91.42%, ± 3.79%, 73.85% ± 7.03%, 48.70% ± 5.30%, and 25.68% ± 4.55%, respectively, and the differences between the groups were statistically significant (F = 6.333, 15.400, 23.700, P < 0.01). The relative expression levels of CD86 mRNA in the normal control group, model group, and pretreatment model group were 1.00 ± 0.00, 2.01 ± 0.04, and 4.17 ± 0.14, respectively, and the differences between the groups were statistically significant (F = 33.800, 106.500, P < 0.01). The relative expression levels of CD163 mRNA in the normal control group, the model group, and the pretreatment model group were 1.00 ± 0.00, 0.85 ± 0.01, and 0.65 ± 0.01, respectively, and the differences between the groups were statistically significant (F = 23.360, 55.350, P < 0.01). The percentages of (F4/80(+)CD86(+)) and (F4/80(+)CD163(+)) in the normal control group and model group and pretreatment model group were 10.67% ± 0.91% and 12.60% ± 1.67%, 20.02% ± 1.29% and 8.04% ± 0.76%, and 43.67% ± 2.71% and 5.43% ± 0.47%, respectively, and the differences among the groups were statistically significant (F = 11.130/8.379, 39.250/13.190, P < 0.01). The liver injury scores of the normal control group, the model group, and the pretreatment model group were 0.22 ± 0.08, 1.32 ± 0.36, and 2.17 ± 0.26, respectively, and the differences among the groups were statistically significant (F = 12.520 and 22.190, P < 0.01). Conclusion: Ces1f may be a hepatic inflammatory inhibitory molecule, and its inhibitory effect production may come from the molecule's maintenance of KC polarization phenotypic homeostasis.

Traditional Chinese medicine health care from "long term taking" drugs in Shen Nong's Classic of Materia Medica / 中国中药杂志

"Taking drugs for a long term" is a qualitative expression of medication method based on the efficacy and safety of Chinese medicine, and the study on it is conducive to the full utilization of the efficacy and rational use of drugs. There are 148 drugs that can be taken for a long time recorded in Shen Nong's Classic of Materia Medica, accounting for 41% of the total drugs. This paper analyzed three-grade classification, natural qualities, four properties and five flavors, and efficacy features of the "long-term taking" drugs(LTTD), thus exploring the herbal source of traditional Chinese medicine health care and the rationality of effect accumulation by long-term taking. It was found that there were more than 110 top-grade LTTD in Shen Nong's Classic of Materia Medica, most of which were herbs, with sweet flavor, flat property, and no toxicity. The efficacies were mainly making body feel light and agile(Qingshen) and prolonging life. Eighty-three LTTD were included in the Chinese Pharmacopoeia(2020 edition). In the modern classification, tonic LTTD accounted for the most, followed by damp-draining diuretic LTTD and exterior-releasing LTTD. Twenty LTTD were included in the "List of Medicinal and Edible Products" and 21 were in the "List of Products Used for Health-care Food", involving in various modern health care effects, such as enhancing immunity, assisting in reducing blood lipids, and anti-oxidation. Shen Nong's Classic of Materia Medica is the classic source of traditional Chinese medicine health care, and its medication thought of taking drugs for a long term to accumulate effects has guiding significance for the regulation of sub-health and chronic diseases nowadays. The efficacy and safety of LTTD have been examined in practice for a long time, and some of the drugs are edible, which is unique in the whole cycle of health-care service, especially in line with the health-care needs in the aging society under the concept of Big Health. However, some records in the book are limited by the understanding of the times, which should be scientifically studied according to the Chinese Pharmacopoeia and the related regulations and technical requirements, under the attitude of eliminating falsifications and preserving the truth and keeping the right essence, so as to achieve further improvement, innovation, and development.

Hepatic COX1 loss leads to impaired autophagic flux and exacerbates nonalcoholic steatohepatitis

The mechanisms underlying autophagic defects in nonalcoholic steatohepatitis (NASH) remain largely unknown. We aimed to elucidate the roles of hepatic cyclooxygenase 1 (COX1) in autophagy and the pathogenesis of diet-induced steatohepatitis in mice. Human nonalcoholic fatty liver disease (NAFLD) liver samples were used to examine the protein expression of COX1 and the level of autophagy. Cox1Δhepa mice and their wildtype littermates were generated and fed with 3 different NASH models. We found that hepatic COX1 expression was increased in patients with NASH and diet-induced NASH mice models accompanied by impaired autophagy. COX1 was required for basal autophagy in hepatocytes and liver specific COX1 deletion exacerbated steatohepatitis by inhibiting autophagy. Mechanistically, COX1 directly interacted with WD repeat domain, phosphoinositide interacting 2 (WIPI2), which was crucial for autophagosome maturation. Adeno-associated virus (AAV)-mediated rescue of WIPI2 reversed the impaired autophagic flux and improved NASH phenotypes in Cox1Δhepa mice, indicating that COX1 deletion-mediated steatohepatitis was partially dependent on WIPI2-mediated autophagy. In conclusion, we demonstrated a novel role of COX1 in hepatic autophagy that protected against NASH by interacting with WIPI2. Targeting the COX1-WIPI2 axis may be a novel therapeutic strategy for NASH.

Effects of Pearl Hydrolysate on Hepatic Sinusoidal Endothelial Cell Viability and Capillarization in Liver Fibrosis / 中国医学科学院学报

Objective To study the effect and mechanism of pearl hydrolysate on hepatic sinusoidal capillarization in liver fibrosis. Methods Hepatic sinusoidal endothelial cells (HSEC) and hepatic stellate cells (HSC-LX2) were incubated with Hepu pearl hydrolysate.The proliferation of HSEC and HSC-LX2 was examined by MTT colorimetry.The cell cycle and apoptosis of HSC-LX2 were measured by flow cytometry.The changes of the microstructures such as fenestra and basement membrane of HSEC were observed by transmission electron microscopy. Results The intervention with leptin increased the viability of HSC-LX2 (P=0.041),decreased the viability of HSEC (P=0.004),and caused capillarization signs such as decreased number and diameter of fenestrae and formation of continuous basement membrane.The treatment with pearl hydrolysate at different doses increased and expanded the fenestrae of HSEC (low dose:P=0.020;medium dose:P=0.028;high dose:P=0.032),disintegrated the extracellular basement membrane of HSEC (low dose:P=0.020;medium dose:P=0.028;high dose:P=0.032),decreased the viability of HSC-LX2 (low dose:P=0.018;medium dose:P=0.013;high dose:P=0.009),and induced the apoptosis of HSC-LX2 (low dose:P=0.012;medium dose:P=0.006;high dose:P=0.005).Pearl hydrolysate exerted therapeutic effect on capillarization in a dose-dependent manner (low dose:P=0.020;medium dose:P=0.028;high dose:P=0.032).Moreover,high-dose pearl hydrolysate showed stronger effect on capillarization of hepatic sinuses than colchicine (P=0.034) and salvianolic acid B (P=0.038). Conclusion Hepu pearl hydrolysate can increase the viability of HSEC,restore the area of fenestrae,disintegrate the basement membrane,and decrease the viability and induce the apoptosis of HSC-LX2,demonstrating significant pharmacological effects on the capillarization of HSEC and HSC-LX2.

Efficacy of nutritional intervention and exercise rehabilitation therapy on sarcopenia in elderly patients with type 2 diabetes mellitus / 中华健康管理学杂志

Objective:To observe the effect of nutrition intervention and exercise rehabilitation treatment on the sarcopenia in elderly patients with type 2 diabetes.Methods:It was a cross-sectional study. From March 2019 to September 2020, 101 elderly patients with type 2 diabetes complicated with sarcopenia treated in the Department of Endocrinology of Tianjin Third Central Hospital were enrolled in this study. The patients were divided into four groups with propensity score matching method: sarcopenia education group without nutrition and resistance training group (group A, n=22), simple nutrition intervention group (group B, n=28), nutritional intervention combined with resistance training group (group C, n=27) and nutritional intervention combined with aerobic and resistance training group (group D, n=24). All the patients were intervened for 24 weeks, two patients in groups C and D dropped due to their own reasons. The 25-dihydroxy vitamin D3 (25(OH)D 3), grip strength, muscle mass of the limbs and short physical performance battery (SPPB) scores were measured before and 24 weeks after the intervention in all the participants. Results:After the intervention, the 25(OH)D 3 levels in the B, C, D groups was (33.45±4.05), (33.68±4.69), (34.28±5.58) μg/L, respectively, all were higher than those before the treatment (all P<0.01), and there was no significant differences among the three groups ( P>0.05). The muscle mass in the B, C, D groups after intervention was 5.650 (5.102, 6.658), 6.601 (6.007, 7.156) and 6.520 (6.017, 7.302) kg/m 2, respectively, all were significantly higher than those before the treatment (all P<0.01); the muscle mass in group C and D increased more significantly than that in group B ( P<0.01), but there was no significant differences between group C and D ( P>0.05). After the intervention, the muscle strength in the C and D groups was 20.60 (19.20, 24.55) kg and 21.15 (19.43, 26.63) kg, and the SPPB scores was 8.00 (7.00, 9.00) points and 8.00 (8.00, 9.00) points, respectively, all were higher than those before the intervention (all P<0.01), but there was no significant differences between the two groups(both P>0.05). The SPPB function score in group D was better than that in group C, the difference was statistically significant ( P<0.05). Conclusions:Nutritional intervention can improve the muscle mass in elderly type 2 diabetes patients with sarcopenia. Combined with rehabilitation training, the muscle strength and muscle function of these patients could also be improved, and the improvement of muscle mass is better than that in patients receiving nutritional intervention only. In terms of increasing muscle strength, the two kinds of rehabilitation training are equivalent. If combined with aerobic exercise, it can also improve the muscle function of these patients.

A comparison of the acute physiology and chronic health evaluation Ⅱ score and the trauma-injury severity score for outcome assessment in intensive care unit trauma patients: a meta-analysis / 中华危重病急救医学

Objective:To assess the ability of the acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) and trauma-injury severity score (TRISS) in predicting mortality in intensive care unit (ICU) trauma patients.Methods:Databases of PubMed, Cochrane Library, SinoMed, CNKI were retrieved from January 1980 to December 2020. The ability of the APACHE Ⅱ and the TRISS to predict mortality in the ICU trauma patients was compared in the retrieval literatures. The relevant literatures were screened by two researchers independently. The data of the included literatures were extracted, and the quality of the included literatures was evaluated. MetaDiSc 1.4 software was used to test the heterogeneity among studies. Meta-analysis was performed on diagnostic accuracy indicators and the summary receiver operator characteristics curve (SROC curve) was fitted. The area under SROC curve (AUC) of the two scores was compared. Deek test was used to analyze literature publication bias.Results:Six studies were selected with 4 054 patients involved with medium and high quality. Meta-analysis results showed that APACHE Ⅱ and TRISS had low sensitivity [the pooled sensitivity and 95% confidence interval (95% CI) was 0.48 (0.41-0.55) and 0.51 (0.41-0.62)], high specificity [the pooled specificity and 95% CI was 0.96 (0.93-0.97) and 0.98 (0.95-0.99)], the pooled diagnostic odds ratio ( DOR) and 95% CI was 20 (14-28) and 46 (18-120), and overall good performance in terms of AUC [the AUC and 95% CI was 0.79 (0.75-0.82) and 0.80 (0.76-0.83)] in predicting the prognosis of ICU trauma patients. There was no statistical difference in AUC between the two scores ( Z = 1.542, P > 0.05). Deek funnel plot showed little publication bias. Conclusion:Both APACHE Ⅱ and TRISS scores could accurately predict mortality in ICU trauma patients.

Application of magnetic resonance quantitative score on the assessment of brain maturity in preterm infants with white matter injury / 中国新生儿科杂志

Objective:To study the relationship between white matter injury (WMI) and brain maturity in preterm infants at full-term corrected gestational age (cGA).Methods:A retrospective study was performed in preterm infants [GA≤32 weeks or birth weight (BW) ≤1 500 g] admitted to the neonatal intensive care unit of the First Affiliated Hospital of Xi'an Jiaotong University from January 2017 to August 2018 and the Northwest Women and Children's Hospital from January 2017 to June 2017. The infants received conventional magnetic resonance imaging (MRI) at cGA 37~42 weeks. The infants were assigned into the WMI group and the control group according to the WMI scoring system, including the total maturation scores (TMS) and four sub-item scores.Results:A total of 118 premature infants were enrolled in this study (17 cases in the WMI group and 101 cases in the control group). The GA was (30.3±1.7) weeks, and BW was (1 356±268) g. The proportion of delayed TMS in the WMI group was significantly higher than the control group [58.8%(10/17) vs. 31.7%(32/101), P<0.05]. The TMS of the WMI group were significantly lower than the control group [(10.7±1.8) vs. (11.8±1.5), P<0.05]. The sub-item scores of TMS showed that the myelination [(2.8±0.6) vs. (3.1±0.4), P<0.05] and glial cell migration bands of the WMI group [(1.6±0.4) vs. (2.1±0.6), P=0.004] were significantly lower than the control group and no significant differences existed in cortical folding and involution of germinal matrix tissue scores between the two groups. Conclusions:The brain maturity of preterm infants with WMI is substantially delayed than those without WMI, including delayed myelination and delayed disappearance of glial cell migration bands.

Changes in circadian gene cryptochrome 2 expression in mouse models of psoriasis and HaCaT cells and their underlying mechanisms / 中华皮肤科杂志

Objective:To investigate changes in circadian gene cryptochrome 2 (CRY2) expression in mouse models of psoriasis and HaCaT cells, and to explore underlying mechanisms.Methods:Imiquimod-induced mouse model experiment: 12 C57BL/6 female mice were randomly and equally divided into imiquimod group receiving topical imiquimod treatment for 5 consecutive days and control group receiving no treatment; these mice were sacrificed on day 6, skin tissues were resected from the back of mice, and immunofluorescence staining was performed to determine the CRY2 expression in the epidermis. HaCaT cell transfection experiment: HaCaT cells with small interfering RNA (siRNA) -mediated knockdown of CRY2 served as siRNA-CRY2 group, and siRNA-NC group as control group; 5-ethynyl-2′-deoxyuridine (EdU) staining was performed to evaluate the proliferative activity of the HaCaT cells, real-time fluorescence-based quantitative PCR (qPCR) to determine the mRNA expression of chemokines in the HaCaT cells, and Western blot analysis to determine phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2) . Tumor necrosis factor-α (TNF-α) -stimulated animal and cell experiments: 12 C57BL/6 female mice were randomly and equally divided into TNF-α group subcutaneously injected with TNF-α solution in the ear for 6 days, and phosphate buffered saline (PBS) group subcutaneously injected with the same amount of PBS; the mice were sacrificed on day 7, skin tissues were resected from the ear of mice, and immunofluorescence staining was conducted to determine the CRY2 expression in the epidermis; CRY2-knockdown HaCaT cells stimulated with 50 ng/ml TNF-α for 12 hours served as siRNA-CRY2 + TNF-α group, and siRNA-NC + TNF-α group as control group; qPCR was performed to determine the mRNA expression of chemokines in HaCaT cells in the above groups. Statistical analysis was carried out by using two-independent-sample t test. Results:Immunofluorescence staining showed that the CRY2 protein expression was significantly lower in the mouse dorsal epidermis in the imiquimod group (0.94 ± 0.23) than in the control group (2.30 ± 0.25, t = 3.99, P = 0.016) . Compared with the siRNA-NC group, the siRNA-CRY2 group showed significantly increased proportions of EdU-positive cells (48.13% ± 10.97% vs. 38.23% ± 0.81%, t = 5.00, P = 0.007) , mRNA expression levels of chemokines CXCL1 and CXCL8, as well as significantly increased phosphorylated (p) -ERK1/2 protein expression levels (all P < 0.05) , while there were no significant differences in the CCL20 mRNA expression or ERK1/2 protein expression between the two groups (both P > 0.05) . Immunofluorescence staining showed significantly decreased CRY2 protein expression level in the mouse ear epidermis in the TNF-α group (0.37 ± 0.34) compared with the PBS group (2.04 ± 0.17, t = 4.38, P = 0.012) ; the relative mRNA expression levels of chemokines CXCL1, CXCL8, and CCL20 in HaCaT cells were significantly higher in the siRNA-CRY2 + TNF-α group than in the siRNA-NC + TNF-α group (all P < 0.05) . Conclusion:CRY2 was markedly underexpressed in psoriasis, which might promote the proliferation of keratinocytes and expression of chemokines CXCL1, CXCL8 and CCL20, and TNF-α might be an upstream cytokine that could downregulate CRY2 expression.

Tumor necrosis factor α-mediated low expression of fatty acid desaturase 2 in psoriasis / 中华皮肤科杂志

Objective:To investigate the expression of fatty acid desaturase 2 (FADS2) in psoriatic skin lesions, as well as its regulatory factors.Methods:FADS2 expression in psoriatic skin lesions was analyzed by using the dataset GDS4602 in Gene Expression Omnibus (GEO) database. Skin tissues were obtained from the back of 5 C57BL/6 mouse models of imiquimod-induced psoriasis, normal skin of 4 patients without psoriasis or other immune skin diseases, lesions of 4 patients with psoriasis before and after 10-week treatment with infliximab, as well as lesions of 3 patients with psoriasis before and after 12-week treatment with secukinumab in Shanghai Skin Disease Hospital. FADS2 expression was determined by both immunohistochemical staining and Western blot analysis in the epidermis of mouse skin tissues, and by immunohistochemical staining in that of human skin tissues. In vitro cultured human immortalized keratinocytes (HaCaT) were divided into several groups to be treated with 50 ng/ml tumor necrosis factor-α (TNF-α) alone for 0, 6, 12 and 24 hours respectively, 200 ng/ml interleukin-17A (IL-17A) alone for 0, 6 and 12 hours respectively, or treated with 50 ng/ml TNF-α and 5 μmol/L BAY 11-7082 (a nuclear factor-κB pathway inhibitor) for 6 hours (TNF-α+ BAY 11-7082 6 h group) , and the cells receiving normal culture served as the control group. After the above treatment, real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were conducted to determine the mRNA and protein expression of FADS2 respectively. Statistical analysis was carried out by using one-way analysis of variance and t test. Results:Analysis of the dataset GDS4602 showed that the FADS2 mRNA expression was significantly lower in the lesional and non-lesional skin tissues from the patients with psoriasis (0.656 ± 0.475, 1.503 ± 1.062, respectively) than in the normal skin tissues (2.035 ± 1.226; F = 55.17, 3.07, P < 0.001, = 0.012, respectively) , and was significantly lower in the lesional skin tissues than in the non-lesional skin tissues from the patients with psoriasis ( F = 26.27, P < 0.001) . Western blot analysis and immunohistochemical staining both showed significantly decreased FADS2 protein expression in the mouse skin tissues in the imiquimod group (gray-value ratio: 0.463 ± 0.172; fluorescence intensity: 21.840 ± 3.125) compared with the normal control group (gray-value ratio: 1.000, t = 7.00, P = 0.002; fluorescence intensity: 30.720 ± 6.850, t = 3.15, P = 0.035) . Compared with the skin lesions before treatment, the FADS2 protein expression significantly increased in the skin lesions from the patients with psoriasis after 10-week treatment with infliximab (43.775± 3.342 vs. 27.950 ±1.218, t = -6.95, P = 0.006) , but was not significantly changed in the skin lesions from the patients with psoriasis after 12-week treatment with secukinumab (28.667 ± 3.402 vs. 31.933 ± 2.987, t = 2.72, P = 0.113) . qPCR revealed that the FADS2 mRNA expression significantly decreased in HaCaT cells in the TNF-α 6 h group and TNF-α 12 h group compared with the TNF-α 0 h group ( P = 0.002, 0.003, respectively) , while there was no significant change in the FADS2 mRNA expression in the IL-17A 6 h group and IL-17A 12 h group compared with the IL-17A 0 h group ( P = 0.849, 0.961, respectively) . The FADS2 mRNA expression significantly decreased in HaCaT cells in the TNF-α 6 h group (0.682 ± 0.132) compared with the control group (1.000, t = 4.82, P = 0.017) , but significantly increased in the TNF-α + BAY 11-7082 6 h group (1.541 ± 0.525) compared with the TNF-α 6 h group ( t = -3.58, P = 0.037) . Western blot analysis revealed significantly decreased FADS2 protein expression in HaCaT cells in the TNF-α 24 h group compared with the TNF-α 0 h group ( F = 6.24, P = 0.013) . Conclusion:FADS2 expression was downregulated in psoriatic lesions, which may be related to TNF-α.

The role and mechanism of sodium valproate in alleviating cardiac and cerebral injuries after cardiopulmonary resuscitation in pigs / 中华急诊医学杂志

Objective:To investigate the role and mechanism of sodium valproate (VPA) in cardiac and cerebral injuries after cardiopulmonary resuscitation (CPR) in pigs.Methods:Twenty-five healthy male domestic pigs, weighing (37±3) kg, were randomly divided into the sham group ( n=6), CPR group ( n=10), and CPR+VPA group ( n=9). Cardiac arrest was induced by alternating current delivered via a pacing catheter in the right ventricle and untreated for 9 min, and then CPR was performed for 6 min, in which this procedure was used to establish the animal model of cardiac arrest and CPR. At 5 min after resuscitation, a dose of 150 mg/kg of VPA was infused with a pump via the femoral vein in 1 h in the CPR+VAP group. At 1 h, 2 h, 4 h and 24 h after resuscitation, blood samples were drawn from the femoral vein, and then used to measure the serum concentrations of cardiac troponin I (cTnI), creatine kinase MB (CKMB), neuron specific enolase (NSE), and S100B protein (S100B) by ELISA. At 24 h after resuscitation, the animals were euthanized, and then tissue specimens in the left myocardium and brain cortex were rapidly harvested to detect the expression levels of C/EBP homologous protein (CHOP), caspase 12, and caspase 3 by Western blot, and the rate of apoptotic cells was detected by TUNEL. Continuous variables were compared with one way analysis of variance among the three groups. Results:(1) After resuscitation, cardiac and cerebral injury biomarkers including cTnI, CKMB, NSE, and S100B in serum were significantly increased in the CPR and CPR+VPA groups compared with the Sham group (all P<0.05). The serum concentrations of cTnI and NSE starting 1 h after resuscitation and the serum concentrations of CKMB and S100B starting 2 h after resuscitation were significantly decreased in the CPR+VPA group compared to the CPR group (all P<0.05). (2) Those proteins related to cell apoptosis mediated by endoplasmic reticulum stress, including CHOP, caspase 12, and caspase 3, were significantly increased, and meanwhile apoptosis index was markedly elevated after resuscitation in the CPR and CPR+VPA groups compared with the Sham group (all P<0.05). Nevertheless, the expression levels of CHOP, caspase 12, and caspase 3 were significantly decreased, and cell apoptosis was markedly reduced in the heart and brain after resuscitation in the CPR+VPA group compared to the CPR group (all P<0.05). Conclusions:VPA can alleviate cardiac and cerebral injuries after CPR in pigs, and its mechanism may be possibly related to the inhibition of cell apoptosis mediated by endoplasmic reticulum stress.

Analysis of articles on dental pulp biology research in China published in international journals from 2011 to 2020 / 中华口腔医学杂志

Objective: To analyze the articles on research of dental pulp biology in China and to understand the situation of China in the entire field of dental pulp biology around the world in order to provide references for further in-depth research in dental pulp biology in China. Methods: Based on Web of Science core collection database, the articles published in the international journals in the field of dental pulp biology from 2011 to 2020 were retrieved and identified. Six indicators including research scale, academic influence, high-ranking and the high-influence journals, research areas, international partnership and project funding were statistically analyzed. Results: Totally 1 215 articles were published by Chinese researchers, which is the most in the research field of dental pulp biology. The total number of citations was 18 328, however the average number of citations of above mentioned articles was slightly lower than that of the world average. The number of articles published in Natural Index Journals and Journal of Dental Research is lower than that of the United States. The research areas of dental pulp biology in China were not only in dentistry, oral surgery and medicine, but also in cell biology, experimental medicine, materials science, engineering, molecular biology and applied microbiology. Articles of internationally cooperated researches were scarce. Most of the researches of dental pulp biology field conducted in China were supported by the National Natural Science Foundation of China. Conclusions: Although there were fruitful research outcomes in the field of dental pulp biology from 2011 to 2020, the academic influence of these researches still needed to be improved. It was recommended that great efforts should be made in developing interdisciplinary, inter-unit and international cooperation, focusing on hotspot and major projects, actively applying for and using of project fundings in order to produce more high-quality research outcomes.

Research progress of small molecule proteasome activators / 药学学报

Proteasome controls the degradation of proteins closely related to life activities and plays a key role in the maintenance of protein homeostasis. Proteasome activities decrease with aging, followed by the overwhelming production of damaged proteins which far exceed the protein consumption. Accumulation of these proteins leads to various diseases including neurodegenerative diseases. Therefore, inducing toxic protein degradation is considered as a promising solution for the treatment of these diseases, while increasing the activity of proteasome is considered as an important strategy. However, the research in this field is still in the preliminary stage, and this review will focus on the discussion of the research progress of various small molecule proteasome activators, including research methods, pharmacological effects, structure-activity relationships and the existing problems.

Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

The use of two inhibitors of Mek1/2 and Gsk3β (2i) promotes the generation of mouse diploid and haploid embryonic stem cells (ESCs) from the inner cell mass of biparental and uniparental blastocysts, respectively. However, a system enabling long-term maintenance of imprints in ESCs has proven challenging. Here, we report that the use of a two-step a2i (alternative two inhibitors of Src and Gsk3β, TSa2i) derivation/culture protocol results in the establishment of androgenetic haploid ESCs (AG-haESCs) with stable DNA methylation at paternal DMRs (differentially DNA methylated regions) up to passage 60 that can efficiently support generating mice upon oocyte injection. We also show coexistence of H3K9me3 marks and ZFP57 bindings with intact DMR methylations. Furthermore, we demonstrate that TSa2i-treated AG-haESCs are a heterogeneous cell population regarding paternal DMR methylation. Strikingly, AG-haESCs with late passages display increased paternal-DMR methylations and improved developmental potential compared to early-passage cells, in part through the enhanced proliferation of H19-DMR hypermethylated cells. Together, we establish AG-haESCs that can long-term maintain paternal imprints.

Clinical observation of orthokeratology with reducing back optic zone diameter in the treatment of myopia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To discuss the efficacy and safety of orthokeratology with reducing back optic zone diameter(5mm-BOZD)compared with conventional back optic zone diameter(6mm-BOZD)in the treatment of adolescent myopia.METHODS: A prospective randomized controlled trial was performed. There were 100 cases with 100 eyes of adolescent myopia(all right eye data were taken)selected from April 2016 to January 2019, the spherical degree was -1.00--5.00D. Then they were randomly divided into the two groups. The experimental group wore 5mm-BOZD orthokeratology, and the control group wore 6mm-BOZD orthokeratology. Their axis length(AL), spherical equivalent(SE), relative peripheral refraction(RPR), best corrected visual acuity(BCVA), uncorrected near visual acuity(NVA), Efron grading was applied to record the anterior segment of the eyes, corneal hysteresis(CH), corneal resistance factor(CRF), corneal-compensated intraocular pressure(IOPcc), average noninvasive Keratograph tear breakup time(NIKBUTav)and higher order aberration(RMSh) were compared between the two groups during the 1a treatment period.RESULTS: After 1a of treatment, the AL in experimental group increased 0.05±0.05mm,while it increased 0.15±0.05mm(t=-8.949, P<0.001)in control group. The SE in experimental group increased -0.18±0.27D,while it increased-0.42±0.35D(t=3.609, P=0.001)in control group. There were statistical differences in RPR changes at N30°, N20° and T30°sites between the two groups(P<0.05). There were no statistical differences in BCVA,NVA,Efron grade,CH,CRF,IOPcc,NIKBUTav and RMSh between the two groups(P>0.05).CONCLUSION:Reducing back optic zone diameter orthokeratology can correct adolescent myopia safely and more effectively during the observation period.

Preparation of magnetic lidocaine microsphere and effects on sciatic nerve block in rabbits / 中华麻醉学杂志

Objective:To develop a novel sustained-release local anesthetic microspheres and evaluate the effects on sciatic nerve block in rabbits.Methods:The magnetic lidocaine microspheres were prepared by W 1/O/W 2 compound emulsion method, investigating their external morphology, measuring the magnetic response characteristics by the VSM and draw the hysteresis loop.The encapsulation efficiency and drug-loading rate were calculated, and the cumulative release curves in vitro were drawn.Fifteen healthy rabbits (half male and half female), aged 5-6 months, weighing 3.0-3.5 kg, were selected for sciatic nerve block and divided into 3 groups ( n=5 each) using a random number table method: magnetic response lidocaine microspheres group (PL group), normal saline control group (C group) and lidocaine group (L group). Magnetic response lidocaine microsphere buffer 2 ml, normal saline 2 ml and 2% lidocaine 2 ml were injected around the rabbit sciatic nerve through a catheter in PL, C and L groups, respectively.The applied magnetic field was withdrawn at 60 h after injection.Before injection (T 0) and at 30 min and 2 , 8, 16, 24, 48, 60, 62 and 64 h after injection (T 1-9), the compound action potentials and conduction velocities of bilateral sciatic nerve trunks were measured, and block was assessed using toe reflex score and modified Tarlov score. Results:The magnetic lidocaine microspheres were brown in color and observed as monodisperse, regular spheres with a diameter of (9±3) μm, an encapsulation rate of 46.18%, a drug loading of 6.02%, and a superparamagnetic release rate of 97% in vitro at 60 h. The hysteresis loop passed through the origin and no hysteresis occurred with the absence of an external magnetic field.Compared with C group, the action potentials and conduction velocities of the sciatic nerve, toe reflex score and modified Tarlov score were significantly decreased at T 1-T 8 in PL group ( P<0.05). Compared with L group, the action potentials and conduction velocities of the sciatic nerve were significantly increased at T 1, the action potential was decreased at T 2-T 8, the conduction velocity was decreased at T 3-T 8, the toe reflex score was increased at T 1 and decreased at T 3-T 8, and the modified Tarlov score was increased at T 1 and T 2 and decreased at T 3-T 8 in PL group ( P<0.05). Conclusions:Magnetic response lidocaine microsphere is successfully developed with good magnetic responsiveness and release and can prolong the sciatic nerve block time in rabbits.

Cost accounting for diagnosis items based on virtual standardized clinical chemistry laboratory / 中华医院管理杂志

Objective:Cost accounting for its diagnosis items based on virtual standardized clinical chemistry laboratory.Methods:Relevant data of clinical chemistry laboratories from January to June 2019 were extracted from the laboratory information systems of 10 hospitals in Shanghai, and three health economic experts and the directors of their laboratory departments were interviewed in this regard.On such basis, a virtual standardized clinical chemistry laboratory was constructed. The project cost of the virtual laboratory was calculated from the aspects of supplies exhaust, labor and others. The routine clinical chemistry diagnosis items were clustered according to the principle of laboratory methods, and the cost differences of items in the same cluster were compared using paired t test. Results:The cost of rate method and dry chemical method in testing alanine aminotransferase was 5.12 and 11.63 respectively, and that of immune turbidimetry and immune scattering turbidimetry method in testing immunoglobulin G was 20.00 and 22.26 respectively. Cluster analysis was conducted on 214 routine clinical biochemical diagnostic items, of which 202 items were classified into 42 clusters. The average of clinical chemistry items accounted for 91.7%(4 493/4 900)of the total per day. Based on enzymology, the calculation costs of alanine aminotransferase(rate method), aspartate aminotransferase(rate method), cholesterol(enzyme method)and uric acid(enzyme method)was 5.12, 5.10, 5.24 and 5.14 respectively, presenting no statistical difference( P>0.05). Conclusions:Research on the cost accounting method of clinical chemistry laboratory diagnosis items constructed includes labor cost, reflects the technical labor value of medical staff. Cost accounting based on project clustering can provide references for medical service pricing and financial management of hospitals.

Analysis of Rancidity Mechanism of Armeniacae Semen Amarum During Storage / 中国实验方剂学杂志

Objective:To explore the mechanism of rancidity during storage by researching the changes of water content， relative permeability of cell membrane and rancidity levels of Armeniacae Semen Amarum in deterioration process. Method:Armeniacae Semen Amarum samples under different storage conditions were evaluated and classified by sensory assessors， and samples with different levels of rancidity were obtained. Water content was measured by toluene method， and water activity was obtained by water activity meter. Malondialdehyde （MDA） and relative conductivity were measured using thiobarbituric acid colorimetry and conductivity meter， respectively. The content of fatty oil was obtained by Soxhlet extraction. The acid value and peroxide value were measured in accordance with the general rules 0713 and 2303 of the 2020 edition of <italic>Chinese Pharmacopoeia</italic> （part Ⅳ）， respectively. Based on the above experimental data， chemometric methods （cluster analysis， principal component analysis） were selected to establish classification and discriminant models of Armeniacae Semen Amarum with different rancidity levels， in order to verify the accuracy of the classification results. Result:According to the results of sensory evaluation， Armeniacae Semen Amarum samples were divided into three classes， including no rancidity， slight rancidity and rancidity. Compared with the no rancid samples， the color of surface and cotyledon were deepened in rancid samples， and the oil was appeared on surface with rancid taste. The values of water content， water activity， MDA content and relative conductivity were all significantly increased in deterioration process （<italic>P</italic><0.01）. The content of fatty oil was significantly decreased with the occurrence of rancidness （<italic>P</italic><0.01）， while the acid value and peroxide value increased significantly （<italic>P</italic><0.01）. The results of cluster analysis and principal component analysis showed that the rancid samples could be distinguished from the no rancid and slightly rancid samples. Conclusion:The storage conditions under high temperature and high humidity can accelerate the rancidness of Armeniacae Semen Amarum， which is accompanied by the increase of internal water content， the increase of cell membrane permeability and the occurrence of fatty acid rancidity. It is suggested that Armeniacae Semen Amarum should be stored in low temperature， dry environment， as well as short storage time.