Bone protective effect of sinomenine against monosodium iodoacetate induced knee and hip injury in rat model: an inflammatory pathway

Purpose: Osteoarthritis (OA) is a degenerative joint disease which is categorized via destruction of joint cartilage and it also affects the various joints, especially knees and hips. Sinomenine active phytoconstituents isolated from the stem of Sinomenium acutum and already proof anti-inflammatory effect against the arthritis model of rodent. In this experimental protocol, we scrutinized the anti-osteoarthritis effect of sinomenine against monosodium iodoacetate (MIA) induced OA in rats. Methods: MIA (3 mg/50 µL) was used for inducing the OA in the rats, and rats received the oral administration of sinomenine (2.5, 5 and 7.5 mg/kg body weight) up to the end of the experimental study (four weeks). The body and organs weight were estimated. Aggrecan, C-terminal cross-linked telopeptide of type II collagen (CTX-II), glycosaminoglycans (GCGs), monocyte chemoattractant protein-1 (MCP-1), Interferon gamma (IFN-γ), antioxidant, inflammatory cytokines, inflammatory mediators and matrix metalloproteinases (MMP) were analyzed. Results: Sinomenine significantly (P < 0.001) boosted the body weight and reduced the heart weight, but the weight of spleen and kidney remain unchanged. Sinomenine significantly (P < 0.001) reduced the level of nitric oxide, MCP-1 and improved the level of aggrecan, IFN-γ and GCGs. Sinomenine remarkably upregulated the level of glutathione, superoxide dismutase and suppressed the level of malonaldehyde. It effectually modulated the level of inflammatory cytokines and inflammatory mediators and significantly (P < 0.001) reduced the level of MMPs, like MMP-1, 2, 3, 9 and 13. Conclusions: Sinomenine is a beneficial active agent for the treatment of OA disease.

Clinical features of acute macular neuroretinopathy associated with COVID-19 / 中华眼底病杂志

Objective:To evaluate the clinical features of acute macular neuroretinopathy (AMN) associated with COVID-19.Methods:A retrospective case series studies. A total of 12 eyes of 8 patients diagnosed of AMN associated with COVID-19 at Peking University People’s Hospital from December 5, 2022 to January 5, 2023 were included. Of the 8 patients, 2 were male (4 eyes) and 6 were female (8 eyes), with an average age of (29.38±8.60) years. All patients underwent best-corrected visual acuity (BCVA), spectral-domain optical coherence tomography (OCT), and infra-red fundus photography (IR). After definite diagnosis, the patients were given symptomatic treatment such as local vasodilation, anti-inflammatory and systemic circulation improvement and nutritional nerve. Follow-up time was 21-30 days weeks. Clinical manifestations, OCT and IR image characteristics, and treatment outcomes were retrospectively analyzed.Results:The time from diagnosis of COVID-19 to the onset of ocular symptoms was (3.00±0.93) days. Among 12 eyes, 6 had complaints of paracentral scotoma, with 2 of them accompanied by visual acuity loss; and 6 had complaints of dark shadows in the vision, with 2 of them accompanied by visual acuity loss. At the initial examination, 2 eyes had a BCVA of less than 0.05, 2 eyes had a BCVA between 0.4 and 0.6, and 8 eyes had a BCVA between 0.8 and 1.0. At the last follow-up, visual symptoms improved in 7 eyes and remained unchanged in 5 eyes. Fundus color photography showed reddish-brown lesions in the macular area. Spectral-domain OCT revealed localized thickening and strong reflection of the outer plexiform layer (OPL) in the macular area, patchy strong reflections in the outer nuclear layer (ONL), and varying degrees of local discontinuity in the adjacent external limiting membrane, ellipsoid zone/interdigitation zone (EZ/IZ), with reduced local reflection in the adjacent retinal pigment epithelium layer in 2 eyes. The strong reflection area of the ONL on corresponding structural OCT was observed more clearly as a lesion range with strong reflection on en-face OCT. The incomplete structure of the EZ/IZ band was observed more clearly as a lesion range with weak reflection on en-face OCT. IR showed several clear-bordered and weakly reflecting lesions at the center of the macula, with the tip pointing to the fovea.Conclusions:AMN associated with COVID-19 tends to occur in young females. The OCT findings of AMN are characterized by strong reflections in the OPL and ONL, and lesion ranges can be observed more clearly at different levels using en-face OCT. The lesions on IR appear as weak reflections.

Effect of intraoperative cell salvage on the number and viability of cancer cells in salvaged autologous blood from patients undergoing liver cancer surgery / 中华麻醉学杂志

Objective:To evaluate the effect of intraoperative cell salvage (ICS) on the number and viability of cancer cells in salvaged autologous blood from the patients undergoing liver cancer surgery.Methods:Twenty patients undergoing open radical primary hepatocellular carcinoma were selected, and blood from the operative field was collected after exposing the liver and treated with ICS. Blood specimens 20 ml from the surgical field (S 1), blood specimens 20 ml before ICS treatment-leukocyte depletion filter (LDF) filtration (S 2) and blood specimens 20 ml after LDF filtration (S 3) were collected and enriched, of which the blood sample 10 ml was used for cancer cell identification and count by immunofluorescence staining, and the remaining blood sample 10 ml was continuously cultured for 3 weeks, and then cell viability was observed by immunofluorescence method. Results:Hepatocellular carcinoma(HCC) cells were identified in 19 S 1 specimens, 18 S 2 specimens, and 16 S 3 specimens, but there was no significant difference in the detection rate among the three specimens ( P>0.05). Compared with S 1 specimens, HCC cell count was significantly reduced in S 2 and S 3 specimens ( P<0.05). There was no significant difference in the HCC cell count between S 3 specimens and S 2 specimens ( P>0.05). After 3 weeks of culture, the results of light microscopy showed that: hepatocellular carcinoma cell clusters were found in S1 specimens, and no hepatocellular carcinoma cell cluster was found in S 2 and S 3 specimens; the results of fluorescence microscopy showed that: 400 and 14 mixed epithelial-mesenchymal HCC cells and 100 and 21 mesenchymal HCC cells were found in S 1 and S 2 specimens, respectively, while no HCC cells were identified in S 3 specimens, among which HCC cells mainly presented as clusters of hepatocellular carcinoma cells in S 1 specimen, while no clusters of hepatocellular carcinoma cells were found in S 2 and S 3 specimens. Conclusions:After treatment with ICS or ICS-LDF, the number and viability of hepatocellular carcinoma cells in salvaged autologous blood are significantly reduced, and hepatocellular carcinoma cells exist as single cells and fail to develop clusters of hepatocellular carcinoma cells; LDF can reduce the risk of hepatocellular carcinoma cell autotransfusion to a certain extent, although it can not effectively filter out hepatocellular carcinoma cells continuously.

Diagnostic Value of 18F-labeled PSMA PET/CT for Regional Lymph Node Metastasis in Prostate Cancer: A Meta-analysis / 肿瘤防治研究

Objective To evaluate the diagnostic value of 18F-labeled PSMA PET/CT for regional lymph node metastasis in prostate cancer. Methods We searched PubMed, Embase, Cochrane Library, Web of Science, CNKI, VIP and Wanfang database from January 1, 2000 to May 31, 2021 for the studies about the diagnosis of 18F-labeled PSMA PET/CT for regional lymph node metastasis and staging in prostate cancer. Two investigators screened literature, extracted relevant data and assessed the quality of the literature independently. The meta-analysis was performed using Meta-disc 1.4 and Stata 16.0 software. Results A total of 8 studies were finally included for the analysis, consisting of 754 prostate cancer patients and 2101 lymph nodes. The results of this meta-analysis showed pooled sensitivity, pooled specificity, pooled positive likelihood ratio, pooled negative likelihood ratio and diagnostic ratio were 0.82 (95%CI: 0.61-0.93), 0.98 (95%CI: 0.91-1.00), 45.7 (95%CI: 9.0-231.3), 0.18 (95%CI: 0.07-0.45) and 251 (95%CI: 34-1851), respectively. The area under the SROC curve was 0.97 (95%CI: 0.95-0.98). Conclusion 18F-labeled PSMA PET/CT has a high diagnostic value for regional lymph node metastasis in prostate cancer.

Strategies and skills of reoperation for recurrent carcinoma at anastomotic stoma after total gastrectomy / 中华普通外科杂志

Objective:To evaluate the strategy and skills of reoperation for recurrent carcinoma at the stoma of esophagojejunostomy after total gastrectomy.Methods:The management experience in 18 patients at the Department of General Surgery, Tumor Hospital of Zhengzhou University from Oct 2016 to Jul 2021 were analyzed retrospectively.Results:Thirteen patients underwent left thoracoabdominal combined incision to completely remove the tumor.One patient developed anastomotic leakage, two patients developed pulmonary infection, one patient developed costochondritis, all were cured and discharged after conservative treatment; Nine patients had no postoperative complications. Two patients abandoned surgery due to high position of the tumor, 3 patients due to poor cardiopulmonary function, and were given radiotherapy and chemotherapy.The postoperative follow-up period was 4.6 to 42.9 months. Four patients died of tumor recurrence, and the remaining patients survived until the end of the follow-up. The overall 1-and 3-year recurrence-free survival rates were 100% and 20%, respectively.Conclusion:Surgery on recurrent carcinoma at esophagojejunostomy stoma after total gastrectomy, although difficult and challenging,could still benefit most patients when at the hands of expertise.

Effects of body mass index on lung function in patients with chronic obstructive pulmonary disease / 中华健康管理学杂志

Objective:To investigate the effects of body mass index (BMI) on lung function in patients with chronic obstructive pulmonary disease (COPD).Methods:A total of 3 312 patients with COPD were selected from outpatients and inpatients in Department of Gerontal Respiratory Medicine of the First Hospital of Lanzhou University from August 2016 to August 2020, including 1 103 patients in stable period and 2 209 patients in acute exacerbation period. According to body mass index (BMI), these COPD patients were divided into four groups: low weight (56 cases, 131 cases), normal weight (448 cases, 945 cases), overweight (424 cases, 773 cases) and obesity groups (175 cases, 360 cases) respectively in stable stage and in acute exacerbation stage. The lung function of inspiratory capacity (IC), vital capacity (VC), residual volume (RV)/total lung capacity (TLC), forced expiratory volume in 1 second (FEV 1), forced vital capacity (FVC), FEV 1/FVC, maximal mid-expiratory flow (MMEF), diffusing capacity of the lung for carbon monoxide (DLCO), DLCO normalized per liter alveolar volume (DLCO/VA), respiratory impedance (Zrs), respiratory resistance at 5 Hz (R5), respiratory resistance at 20 Hz (R20) and respiratory reactance at 5 Hz (X5) were measured using MasterScreen PFT in all patients, and the influence of BMI on lung function was analyzed respectively. The risk factors of impaired lung function were analyzed by ordered logistic regression with lung function grade as dependent variable and age, gender, smoking history, smoking index and BMI as independent variables (“%pred”represents the percentage of predicted value). Results:The proportion of patients with lung function grade Ⅲ/Ⅳ in acute exacerbation period (64.9%, 37%, 27.4%, 24.4%) was higher than that in stable stage (42.9%, 25.9%, 13.7%, 9.1%), while the proportion of patients with lung function grade Ⅰ in stable stage (21.4%, 34.2%, 38.2%, 40.0%) was higher than that in acute exacerbation period (7.6%, 20.0%, 25.4%, 22.8%) (all P<0.05). The IC%pred, VC%pred, FEV 1%pred, FVC%pred, FEV 1/FVC, MMEF%pred, DLCO%pred, DLCO/VA%pred and R20 in low weight group were significantly lower than other groups both in stable period and acute exacerbation period (all P<0.05). The RV/TLC was higher in low weight group than that of normal weight and overweight groups in both periods (all P<0.05). The IC%pred, FEV 1%pred, FEV 1/FVC, DLCO%pred, DLCO/VA%pred, R5 and R20 in overweight and obesity groups were higher than those of normal weight group (all P<0.05). The RV/TLC, FEV 1/FVC, DLCO%pred, DLCO/VA%pred, Zrs, R5 and R20 in obesity group were higher than those of overweight group (all P<0.05). The ordered logistic regression analysis showed that low weight was independent risk factor for impaired lung function of COPD both in stable period [ OR (95% CI) 2.316 (1.206-3.554)] and acute exacerbation period [ OR (95% CI): 2.457 (1.647-3.669)]. Conclusion:Lower BMI has an adverse effect on lung function, and it is an independent risk factor for impaired lung function in COPD patients.

Comparison of long-term macular morphology after laser retinal photocoagulation and anti-VEGF therapy and their effects on vision prognosis for type 1 ROP / 中华实验眼科杂志

Objective:To evaluate and compare the foveal microvascular morphology and central foveal thickness (CFT) after laser retinal photocoagulation and intravitreal injection of anti-vascular endothelial growth factor (VEGF) in patients with retinopathy of prematurity (ROP), and to explore the factors affecting the prognosis of vision.Methods:A cohort study was conducted.Forty children (40 eyes) aged 4-6 years, who had been treated in Peking University People's Hospital for type 1 ROP from January 2019 to December 2020, were enrolled.Optical coherence tomography angiography (OCTA), best corrected visual acuity (BCVA) and refractive status of the patients were examined.The patients were divided into laser retinal photocoagulation group and anti-VEGF group according to they received a single laser retinal photocoagulation therapy or a single intravitreal injection of anti-VEGF drugs (conbercept or ranibizumab 0.25 mg/0.025 mL) after birth.Twenty age-matched full-term healthy children (20 eyes) were enrolled as the normal control group.The FAZ area, superficial and deep foveal vessel density (VD) and CFT of the affected eyes were measured by OCTA at 4-6 years after treatment to investigate the influence of gestational age, birth weight, morphological characteristics of foveal microvessels and CFT on the prognosis of BCVA.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Peking University People's Hospital (No.2017PHB179-01). Written informed consent was obtained from the guardians prior to any medical examination.Results:There were statistically significant differences in FAZ area, superficial foveal VD and deep foveal VD among the three groups ( F=12.321, 8.436, 5.497; all at P<0.05). The FAZ area was smaller, and the superficial and deep foveal VD of the laser photocoagulation group and the anti-VEGF group were greater than those in the normal control group, and the difference was statistically significant (all at P<0.05). The CFT of the laser photocoagulation group was (267.6±11.8)μm, greater than (259.5±12.9)μm of the anti-VEGF group and (242.4±12.3)μm of normal control group, and the CFT value of the anti-VEGF group was greater than that of the normal control group, and the differences were statistically significant (all at P<0.05). There was a strong negative correlation between the superficial foveal VD and FAZ area ( r=-0.713, P<0.05), a moderate negative correlation between the deep foveal VD and FAZ area ( r=-0.565, P<0.05), and a moderate positive correlation between gestational age and FAZ area ( r=0.485, P<0.05). Pearson correlation analysis results showed that gestational age, FAZ, superficial foveal VD, deep foveal VD, CFT were all correlated with BCVA (all at P<0.05). The effects of gestational age and FAZ on BCVA were both statistically significant ( R2=0.615, both at P<0.05). Conclusions:The morphology of microvessels in macular fovea and the prognosis of BCVA in the affected eye is similar at 4-6 years after laser retinal photocoagulation and intravitreal injection of anti-VEGF drugs for type 1 ROP.The CFT of the affected eye after anti-VEGF drug therapy is better than those after laser retinal photocoagulation.Gestational age and FAZ are the influencing factors of visual acuity after treatment in children with type 1 ROP.

Mori Folium Improves Glucose and Lipid Metabolism Disorders in Rats with Type 2 Diabetes Mellitus by Regulating PI3K/Akt/PPARα/CPT-1 Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect and mechanism of Mori Folium extract on the glucose and lipid metabolism disorders in the liver of rats with type 2 diabetes mellitus （T2DM） through the phosphatidylinositol 3-kinase/protein kinase B/peroxisome proliferation-activated receptor α/carnitine palmitoyl transferase-1 （PI3K/Akt/PPARα/CPT-1） signaling pathway. MethodThe T2DM model was induced by the high-fat diet combined with the intraperitoneal injection of streptozotocin （STZ）. The model rats were randomly divided into a model group， a metformin （0.2 g·kg-1） group， and a Mori Folium water extract （4.0 g·kg-1） group according to blood glucose and body weight. In the 8-week administration， fasting blood glucose was measured at the same time every week. The histomorphological and fat changes in the rat liver were observed by hematoxylin-eosin （HE） staining and oil red O staining. The levels of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） in the serum were measured by biochemical methods. Western blot （WB） was used to quantitatively detect the protein expression of p-PI3K，PI3K，p-Akt，Akt，PPARα，and CPT-1 in the rat liver. ResultAfter 8-week administration， the blood glucose of rats was higher in the model group than that in the control group （P<0.01）， and lower in the Mori Folium water extract group than that in the model group （P<0.01）. The results of HE staining showed that the liver tissue structure of the control group was complete， and the hepatocytes were arranged radially around the central vein， while the hepatocyte injury in the model group was obvious. Compared with the model group， the Mori Folium water extract group showed improved vacuolar degeneration and no lesions such as small bile duct hyperplasia. Oil red O staining showed that there was no obvious steatosis and necrosis in the hepatocytes of rats in the control group， and no lipid droplets in the hepatocytes were observed， while the model group showed increased lipid droplets. Mori Folium significantly reduced the lipid droplets in the liver. Biochemical analysis showed that the levels of TC， TG， LDL-C， AST， and ALT in the model group were significantly higher than those in control group （P<0.01）. The levels of TC， TG， LDL-C， AST， and ALT in the Mori Folium water extract group were significantly lower than those in the model group （P<0.05，P<0.01）. WB showed that the protein expression of p-PI3K/PI3K， p-Akt/Akt， PPARα， and CPT-1 in the model group were lower than those in the control group （P<0.01）. Mori Folium water extract could increase the protein expression of p-PI3K/PI3K， p-Akt/Akt， PPARα， and CPT-1 （P<0.05 or P<0.01）. ConclusionThe hypoglycemic mechanism of Mori Folium water extract may be related to the regulation of the PI3K/Akt/PPARα/CPT-1 signaling pathway.

Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis / 中国天然药物

Phenylalanine ammonia-lyase (PAL), which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid, is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes. Schisandra chinensis, a woody vine plant belonging to the family of Magnoliaceae, is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity. However, the functional role of PAL in the biosynthesis of lignan is relatively limited, compared with those in lignin and flavonoids biosynthesis. Therefore, it is essential to clone and characterize the PAL genes from this valuable medicinal plant. In this study, molecular cloning and characterization of three PAL genes (ScPAL1-3) from S. chinensis was carried out. ScPALs were cloned using RACE PCR. The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis. In order to determine their catalytic activity, recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli (BL21-DE3), followed by Ni-affinity purification. The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds. The optimal temperature, pH value and effects of different metal ions were determined. Vmax, Kcat and Km values were determined under the optimal conditions. The expression of three ScPALs in different tissues was also determined. Our work provided essential information for the function of ScPALs.

A new norsesquiterpenoid from Schisandra chinensis / 中国中药杂志

This study investigated the chemical components from the leaves and stems of Schisandra chinensis. Three norsesquiterpenoids were isolated from S. chinensis by various column chromatographies(silica gel, Sephadex LH-20, and MCI), reversed-phase medium-pressure preparative, and semi-preparative high-performance liquid chromatography(HPLC). Their structures were identified based on physicochemical properties, mass spectrometry(MS), nuclear magnetic resonance(NMR), ultraviolet(UV), and electro-nic circular dichroism(ECD) as(3R,4R,5R,6S,7E)-3,4,5,6-tetrahydroxy-7-megastigmen-9-one(1),(3S,5R,6R,7E)-3,5,6-trihydroxy-7-megastigmen-9-one(2), and(3S,4R,9R)-3,4,9-trihydroxymegastigman-5-ene(3). Compound 1 was a new compound, and its absolute configuration was determined by ECD. Compounds 2 and 3 were isolated from the Schisandra plant for the first time.

Propagation and phenotypic analysis of mutant rabbits with MSTN homozygous mutation / 生物工程学报

Myostatin gene (MSTN) encodes a negative regulator for controlling skeletal muscle growth in animals. In this study, MSTN-/- homozygous mutants with "double muscle" phenotypic traits and stable inheritance were bred on the basis of MSTN gene editing rabbits, with the aim to establish a method for breeding homozygous progeny from primary MSTN biallelic mutant rabbits. MSTN-/- primary mutant rabbits were generated by CRISPR/Cas9 gene editing technology. The primary mutant rabbits were mated with wild type rabbits to produce F1 rabbits, whereas the F2 generation homozygous rabbits were bred by half-sibling mating or backcrossing with F1 generation rabbits of the same mutant strain. Sequence analysis of PCR products and its T vector cloning were used to screen homozygous rabbits. The MSTN mutant rabbits with 14-19 week-old were weighed and the difference of gluteus maximus tissue sections and muscle fiber cross-sectional area were calculated and analyzed. Five primary rabbits with MSTN gene mutation were obtained, among which three were used for homozygous breeding. A total of 15 homozygous rabbits (5 types of mutants) were obtained (M2-a: 3; M2-b: 2; M3-a: 2; M7-a: 6; M7-b: 2). The body weight of MSTN-/- homozygous mutant rabbits aged 14-19 weeks were significantly higher than that of MSTN+/+ wild-type rabbits of the same age ((2 718±120) g vs. (1 969±53) g, P < 0.01, a 38.0% increase). The mean cross sections of gluteus maximus muscle fiber in homozygous mutant rabbits were not only significantly higher than that of wild type rabbits ((3 512.2±439.2) μm2 vs. (1 274.8±327.3) μm2, P < 0.01), but also significantly higher than that of MSTN+/- hemizygous rabbits ((3 512.2±439.2) μm2 vs. (2 610.4±604.4) μm2, P < 0.05). In summary, five homozygous mutants rabbits of MSTN-/- gene were successfully bred, which showed a clear lean phenotype. The results showed that the primary breeds were non-chimeric mutant rabbits, and the mutant traits could be inherited from the offspring. MSTN-/- homozygous mutant rabbits of F2 generation could be obtained from F1 hemizygous rabbits by inbreeding or backcrossing. The progenies of the primary biallelic mutant rabbits were separated into two single-allelic mutants, both of which showed a "double-muscle" phenotype. Thus, this study has made progress in breeding high-quality livestock breeds with gene editing technology.

Analysis of the association between different treatments and high-risk histopathologic features of bilateral retinoblastomas / 中华眼底病杂志

Objective:To evaluate the pathological features of bilateral retinoblastoma (RB) and the relationship between different treatments and high-risk histopathologic features (HHF).Methods:Retrospective series of case studies. From 1999 to 2018, 73 patients with binocular RB diagnosed by pathological examination in Department of Ophthalmology, Peking University People's Hospital were included in the study. Among them, 50 patients were male (68.5%, 50/73), 23 patients were females (31.5%, 23/73); 11 patitents had a family history of RB. The mean age at the first diagnosis was 14.8±15.6 months. The average time between first diagnosis and first intervention was 3.97±4.74 months. According to the international classification standard of intraocular RB staging, among the 73 eyes, C, D and E stages were 2 (2.7%, 2/73), 15 (20.5%, 15/73), and 56 (76.7%, 56/73) eyes, respectively. Ocular images for each patient were obtained using a wide-angle contact fundus camera during examination under general anaesthesia. The treatment protocol (globe salvaging or enucleation) depended on the result of several clinical features. Globe salvaging treatment included chemotherapy combined with local therapy such as intra-arterial chemotherapy (IAC), intravitreal chemotherapeutics injection, cryotherapy, laser, transpupillary thermotherapy and radiotherapy. If globe salvaging failed, enucleation was offered and histopathologic analysis was conducted of the enucleated eye, the ophthalmic pathologist read and evaluated the presence of HHF. Independent samples t-test was performed to compare the continuous variables. The pathological features and the relationship between different treatments and HHF were analyzed. Group difference was calculated with chi-square. Results:Among the 73 eyes, the first treatment was enucleation in 21 eyes (28.8%, 21/73); 52 eyes (71.2%, 52/73) were treated with eye protection. After enucleation, 9 cases (12.3%, 9/73) had recurrence and metastasis, and 7 cases (9.6%, 7/73) died. The intervention time of patients with recurrence and metastasis and those without recurrence and metastasis were 7.4±7.3 and 3.5±4.1 months respectively; the first intervention time of patients with recurrence and metastasis was significantly later than that of patients without recurrence and metastasis, but the difference was not statistically significant ( t=-1.561, P=0.154). The pathological examination results showed that there were 26 eyes (35.6%, 26/73) with HHF, 4 (26.7%, 15/26) and 22 (39.3%, 22/56) eyes were in stage D and E, respectively. Those who received other treatments before enucleation had lower HHF percentages after enucleation than those who did not receive corresponding treatments, but the difference was not statistically significant ( χ2=1.852, 0.074, 0.000, 1.007, 0.007, 2.729; P>0.05). Among the 26 eyes, 5 (83.3%, 5/6) and 21 (31.3%, 21/67) eyes were treated with systemic chemotherapy combined with and without IAC, respectively, and there was a significant difference in the percentage of HHF ( χ2=4.422, P=0.035). Conclusions:IAC eye-preserving therapy before enucleation has a significant effect on HHF.

Clinical analysis of 40 newborns with head and neck occupying lesions / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the clinical features, diagnoses and treatments of head and neck occupying lesions in newborns. Methods: All newborns with head and neck occupying lesions admitted to Neonatel Intensive Care Unit of the First Affiliated Hospital of Zhengzhou University form January 2014 to November 2019 were included. There were 23 males and 17 females, admission age was from 2 d-28 d, and the clinical manifestations, examinations, treatments and outcomes were evaluated. Results: Among 40 newborns with head and neck occupying lesions, 22 cases were admitted with dyspnea, 15 cases with masses in oral cavity or head and neck, 2 cases with fever as the first symptom, and 1 case with hoarseness as the first symptom. There were 5 cases with local infection. All cases were examined with local ultrasound and CT or MRI. Nine cases with severe dyspnea were treated with invasive ventilationm, of them 6 cases underwent invasive ventilation for more than 48 hours, 4 cases received tracheal intubation and artificial nose. Diagnostic punctures were performed in 2 cases. Seven cases received conservative treatments. Surgeries were performed in 31 cases, and 25 cases obtained pathologic diagnoses, including 3 cases of soft palate mature teratomas, 1 case of hard palate teratoma, 1 case of granulosa cell tumor, 1 case of lobulated spindle cell tumor in tongue base, 1 case of polyp in right glottis, 1 case of polyp at esophageal entrance, 4 cases of lingual root cysts, 1 case of laryngeal cyst, 2 cases of thyroglossal duct cysts, 2 cases of lymphangiomas, 1 case of lymphangioma with hibernoma, 1 case of tracheal cyst, 1 case of esophageal cyst, 3 cases of left neck abscesses, 1 case of occipital hemangioma, and 1 case of left temporoparietal abscess. Conclusions: The head and neck occupying lesions in the newborn is prone to upper airway obstruction. Imaging examination can assist the diagnosis. Different treatments can be selected according to the natures of occupying lesions.

The value of dual-layer detector spectral CT in evaluation of immunohistochemical expression in breast cancer patients / 中华放射学杂志

Objective:To investigate the value of dual-layer detector spectral CT in preoperative evaluation of immunohistochemical expression levels in breast cancer patients.Methods:The clinical, pathological and imaging data of breast cancer patients who received preoperative spectral CT chest scan in Affiliated Hospital of Southwest Medical University from October 2020 to January 2021 were retrospectively analyzed. A total of 70 patients were divided into positive or negative estrogen receptor (ER) group, positive or negative progesterone receptor (PR) group, positive or negative human epithelial growth factor receptor 2 (HER2) group, and positive or negative Ki-67 group according to the immunohistochemistry. Philips workstation and Special Diagnostic Suite software were used for processing and analysis. The CT value and iodine concentration value (IC) of the lesion and the effective atomic number (Z eff) of the lesion in the arterial and venous phases were measured, and the standardized CT value (NCT), standardized iodine concentration value (NIC), slope (K) of the keV-CT value curve in the arterial and venous phases as well as the CT difference and the IC difference in the arteriovenous phase were calculated. Independent sample t test was used to compare the differences in spectral parameters between patients with positive and negative immunohistochemical parameters, and ROC curve was used to analyze the efficacy of spectral parameters in the differential diagnosis of positive and negative different immunohistochemical parameters. Results:Z eff, NIC, K value, CT value, IC value of arterial and venous phase and NCT value of venous phase in negative ER group were higher than those in positive ER group, the difference was statistically significant ( P<0.05). Z eff, NIC, K value, IC, CT difference in arteriovenous phase and IC difference in arteriovenous phase in positive HER2 group were higher than those in negative HER2 patients, the difference was statistically significant (P<0.05). The arterial K value in positive Ki-67 group was higher than that in negative Ki-67 group, the difference was statistically significant ( P<0.05). Taking arterial phase K value of 1.22 as the diagnostic threshold, it had the highest diagnostic efficiency in differential diagnosis of ER positivity versus negativity (area under curve of 0.791, sensitivity of 84.00%, specificity of 65.00%). Taking the arteriovenous phase IC difference of 0.47 mg/ml as the diagnostic threshold, it had the highest performance in differential diagnosis of HER2 positivity versus negativity (area under curve of 0.736, sensitivity of 76.32%, specificity of 65.62%). Conclusion:Dual-layer detector spectral CT has certain diagnostic value for the immunohistochemical expression of breast cancer patients, which can provide reference for clinical diagnosis and treatment.

Construction of point mutation rabbits using CRISPR/Cas9 / 浙江大学学报·医学版

To establish a rabbit model of proprotein convertase subtilisin/kexin type9 () point mutation with CRISPR/Cas9 gene editing technique. According to the PubMed gene protein data, the PCSK9 protein functional regions of human and rabbit were analyzed by Blast. The 386S (Ser) amino acid functional region of human gene was homologous to the 485S of rabbit gene. Three small guide RNAs and one single-stranded donor oligonucleotide were designed according to the 485S base substitution position and sequence analysis of rabbit gene. The synthetic small guide RNAs, Cas9 mRNA and single-stranded donor oligonucleotide were co-injected into the cytoplasm of rabbit fertilized eggs and the embryos were transferred into the pregnant rabbits. PCR, TA cloning and off-target analysis were performed on the F0 rabbits to identify whether the PCSK9 mutation was successful. Fifteen F0 rabbits were obtained. The sequencing results showed that one of them was PCSK9 point mutation homozygote and two of them were PCSK9 point mutation heterozygotes, and the mutation could be stably inherited. The rabbit model of PCSK9 point mutation was successfully constructed by CRISPR/Cas9 technique, which provides an animal model for exploring the molecular mechanism of impaired PCSK9 function and developing reliable and effective diagnosis and treatment measures.

Effect of Guizhitang with Different Proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on Overactive Activation of Cardiac Sympathetic Nervous System in Salt-Sensitive Hypertensive Rats / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of Guizhitang with different proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on overactive cardiac sympathetic nerves in salt-sensitive hypertensive rats. Method:Randomly divide the forty male 6-week-old salt-sensitive hypertensive rats into five groups： the normal control group， the model group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group，and the Cinnamomi Ramulus and Paeoniae Alba Radix 2∶1 group， each group has 8 animals， the normal control group was fed with low-salt feed， and the remaining four groups were fed with 8% high-salt feed. After 4 weeks of feeding， gastric feeding was started. Give both the normal control group and model group saline and the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the 1∶2 group，and the 2∶1 group， were given Guizhitang aqueous solution at 4.0， 5.5 and 5.5 g·kg-1， respectively. Continuous gavage intervention was held for 4 weeks. IITC multi-channel non-invasive sphygmomanometer was used to detect changes of systolic blood pressure before and after treatment in rats. Left ventricular anterior wall end-diastolic thickness （LVAWd） and interventricular septal diastolic thickness （IVSd） were detected by echocardiography. Hematoxylin-eosin（HE）staining and Masson staining were used to observe the myocardial morphological changes of rats in each group， Western blot was used to detect the expression of nerve growth factor （NGF）， growth-associated protein 43 （GAP43） and tyrosine hydroxylase （TH） protein. Result:After 4 weeks of intervention with Guizhitang， compared with the normal control group， the blood pressure， LVAWd and IVSd of the model group were significantly increased， and the expressions of NGF， TH and GAP43 protein were significantly increased （P<0.01）. HE and Masson staining results showed that the model group had myocardial cell edema， a large number of inflammatory cell infiltration， myocardial fiber hyperplasia and disordered arrangement， and a large amount of collagen deposition could be seen in the intercellular substance. Compared with model group， the systolic blood pressure of rats in each Guizhitang group increased slowly， and the expression of NGF， TH and GAP43 protein decreased （P<0.05，P<0.01），the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group has the best effect. The results of echocardiography shows that the 1∶1 Guishao group could reduce LVAWd and IVSd levels （P<0.05，P<0.01）， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group could reduce IVSd level （P<0.05）， there was no statistical difference in LVAWd， there was no statistical difference in LVAWd and IVSd in 2∶1 group. In terms of myocardial morphology， each group of Guizhitang can reduce cell edema and inflammatory cell infiltration， reduce myocardial fiber hyperplasia and collagen deposition， and improve the disorder of myocardial fiber arrangement. Among them， the 1∶1 group has the best effect. Conclusion:Guizhitang can inhibit the overactive activation of cardiac sympathetic nervous system，reduce the extent of myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy， and protect salt-sensitive hypertension rats， whose mechanism may be related to regulating the expression of heart NGF.Among them， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group is better than the 1∶2 and 2∶1 group in reducing myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy.

Clinical Efficacy and Mechanism of Xiao Qinglongtang in Treating Chronic Heart Failure with Cold Phlegm in Lung / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of Xiao Qinglongtang on chronic heart failure with cold phlegm in lung， and explore its mechanism of action. Method:A total of 87 patients with definite chronic heart failure were divided into observation group and control group by random number table method．The two groups received routine western medicine at the same time. Forty-two cases in observation group were treated with Xiao Qinglongtang based on western medicine， and 45 cases in control group received the same dose of placebo. Both groups were treated for 4 weeks， and then their cardiac function， traditional Chinese medicine （TCM） syndrome score and efficacy were compared before and after treatment. Left ventricular ejection fraction （LVEF） and N-terminal pro-brain natriuretic peptide （NT-proBNP） were measured in both groups before and after treatment. The changes of standard deviation of NN intervals （SDNN）， square root of the mean squared differences of successive NN intervals （RMSSD）， high frequency （HF） and low frequency （LF） that reflect autonomic nerve function indexes in heart rate variability （HRV） after treatment were compared between two groups. The changes of inflammatory indicators such as interleukin-6 （IL-6） and highly sensitive C reaction protein （hs-CRP） were detected. Result:After treatment， the total effective rate for cardiac function in observation group was higher than that in control group （P<0.05）. The TCM symptom scores were improved after treatment in both groups （P<0.05）， and the total effective rate in observation group was higher than that in control group （P<0.01）. After treatment， LVEF levels significantly increased （P<0.01） and NT-proBNP levels significantly decreased （P<0.01） in both groups， and the effect in observation group was more obvious （P<0.01）. After treatment， SDNN， RMSD， HF and LF indicators in HRV were all higher than those before treatment in both groups （P<0.01）， and the improvement in observation group was more significant than that in control group （P<0.01）. The levels of IL-6 and hs-CRP decreased after treatment in both groups （P<0.01）， and the level of observation group was significantly lower than that of control group （P<0.01）. Conclusion:Xiao Qinglongtang has certain clinical efficacy in treating chronic heart failure with cold phlegm in lung as it can improve the clinical symptoms of patients， regulate autonomic nervous balance， and inhibit inflammatory factors， providing new clinical ideas to treat chronic heart failure in TCM.

Effect of Guizhitang with Different Proportions of Cinnamomi Ramulus and Paeoniae Alba Radix in Regulating TGF-β1/Smads Signaling Pathway and Chronic Inflammation and Alleviating Myocardial Fibrosis in Salt-sensitive Hypertensive Rats / 中国实验方剂学杂志

<b>Objective::To observe the effect of Guizhitang with different proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on the expressions of transforming growth factor-<italic>β</italic>₁(TGF-<italic>β</italic>₁)/Smads signaling pathway and interleukin-10(IL-10), IL-6 and tumour necrosis factor-<italic>α</italic>(TNF-<italic>α</italic>)related inflammatory cytokines in salt-sensitive hypertensive rats, in order to explore the mechanism of Guizhitang in improving myocardial fibrosis in salt-sensitive hypertensive rats. <b>Method::Totally 40 male 6-week-old salt-sensitive rats were randomly divided into 5 groups: the normal control group, the model group, the 1∶1(RC/peony)Guishao group, the 1∶2 Guishao group, and the 2∶1 Guishao group, with 8 in each group. The normal control group was fed with normal salt diet, while the other four groups were fed with high-salt diet. After 4 weeks of feeding, the rats were given intragastric administration, the normal control group and the model group were given the same amount of normal saline, and the 1∶1 Guishao group, the 1∶2 Guishao group and the 2∶1 Guishao group were given 4.0, 5.5, 5.5 g·kg^-1 of Guizhitang by gavage for 4 weeks. Blood pressure was measured once a week, left ventricular end systolic diameter(LVESD), left ventricular end diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening fraction (LVFS) were detected by using echocardiogram. The pathological changes of myocardial morphology were observed by htoxylin eosin(HE)and Masson staining. The expressions of type Ⅰ and Ⅲ collagen in myocardial tissue of each group was detected by immunohistochemistry. The mRNA expression levels of IL-10, IL-6 and TNF-<italic>α</italic> in myocardial tissue of each group were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR). The protein expression levels of TGF-<italic>β</italic>₁, <italic>α</italic>-smooth muscle actin(<italic>α</italic>-SMA), Smad2, Smad3 and Smad7 in myocardial tissue of each group were detected by Western blot. <b>Result::Compared with the normal control group, the blood pressure was increased in the model group at 8-15 weeks, LVESD, LVEDD were increased in the model group, while LVFS, LVEF were decreased in the model group. The collagen volume fraction was increased, immunohistochemistry showed the expression levels of type Ⅰ and Ⅲ collagen were increased, mRNA expression levels of IL-10, IL-6 and TNF-<italic>α</italic> were increased, the protein expression levels of TGF-<italic>β</italic>₁, Smad2, Smad3 and <italic>α</italic>-SMA were increased, whereas the protein expression of Smad7 was decreased (<italic>P</italic><0.01). Compared with the model group, the blood pressure rise of each group of Guizhitang was delayed in 12-15 weeks, LVESD and LVEDD were decreased in Guizhitang group (<italic>P</italic><0.01), LVFS, LVEF were increased in Guizhitang group (<italic>P</italic><0.01), the collagen volume fraction was decreased in Guizhitang group (<italic>P</italic><0.01), and the expressions of type Ⅰ and Ⅲ collagen were decreased in Guizhitang group (<italic>P</italic><0.01). At the same time, the mRNA expression of IL-10 was increased in Guizhitang group (<italic>P</italic><0.05, <italic>P</italic><0.01), the mRNA expressions of IL-6 and TNF-<italic>α</italic> were decreased in Guizhitang group (<italic>P</italic><0.01), and the protein expressions of TGF-<italic>β</italic>₁, Smad2, Smad3 and <italic>α</italic>-SMA were decreased in Guizhitang group (<italic>P</italic><0.05, <italic>P</italic><0.01), and the protein expression of Smad7 was increased in Guizhitang group (<italic>P</italic><0.01). Compared with the 2∶1 Guishao group, the effect of the 1∶1 Guishao group in improving the above indicators was more obvious (<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::Guizhitang with different proportions of Ramulus Cinnamomi and Poeny can alleviate the degree of myocardial fibrosis in salt-sensitive hypertensive rats. The mechanism may be related to the regulation of TGF-<italic>β</italic>₁/Smads signaling pathway and the reduction of inflammatory response. Besides, the 1∶1 Guishao group showed the optimal effect in reducing inflammation and improving myocardial fibrosis.

Relationship between epicardial adipose tissue based on CT quantification and coronary artery disease: Meta-analysis / 中国介入影像与治疗学

Objective: To observe the correlation between CT quantification of epicardial adipose tissue (EAT) and coronary artery disease (CAD) using meta-analysis. Methods: Case-control studies on the relationship between CT quantification of EAT and CAD in CNKI, Wanfang, VIP, PubMed, Web of science and Medline were searched before December 31, 2019. The standardized mean difference (SMD) with 95%CI between CAD group and non-CAD group was analyzed using Stata 12.0 software. Results: Totally 19 articles (5 129 cases in CAD group and 7 434 cases in non-CAD group) were included. Meta-analysis showed that the thickness (SMD=0.46, 95%CI [0.26, 0.66], P<0.01) and volume (SMD=0.65, 95%CI[0.46, 0.86], P<0.01) of EAT in CAD group were larger than those in non-CAD group. Conclusion: CT quantitative EAT was correlated with CAD.

Cloning of AhWRKY33 from Asarum heterotropoides and its genetic transformation in Arabidopsis / 中国中药杂志

The WRKY family genes, which play an important role in plant morphogenesis and stress response, were selected based on the data of the full-length transcriptome of Asarum heterotropoides. Using AtWRKY33, which regulates the synthesis of the camalexin in the model plant Arabidopsis to compare homologous genes in A. heterotropoides, primers were designed to amplify the open reading frame(ORF) fragment of AhWRKY33 gene by RT-PCR using total RNA of A. heterotropoides leaves as template. Real-time PCR results showed that there was a significant difference between the aerial part and the underground part of A. heterotropoides, the toxic aristolochic acid content is highly expressed in the leaves higher than the root. After verification, the WRKY33 gene of A. heterotropoides is ORF long 1 686 bp, encoding 561 amino acids.AhWRKY33 had two conserved WRKYGQK domains. According to the classical classification, it belongs to group Ⅰ WRKY transcription factor. A. heterotropoides WRKY33 had some homology with amino acids of other species. The study successfully constructed the plant eukaryotic expression vector PHG-AhWRKY33 and transformed Arabidopsis thaliana, the transgenic Arabidopsis was obtained by PCR detection and hygromycin resistant plate screening. It found that the germination of transgenic Arabidopsis seeds was accelerated and the stress resistance was increased. It laid a foundation for further analysis of WRKY transcription factor in the growth and development of A. heterotropoides and the synthesis of secondary metabolites.