ObjectiveTo observe the effect of Yangxin Tongmai Formula （养心通脉方） by midnight-noon ebb-flow administration method for rat models of premature coronary heart disease （PCHD） with blood stasis syndrome， and to explore the possible mechanism of action from the perspective of DNA methylation differential gene expression. MethodsThere were 3 SD rats in each of the blank group， model group and Yangxin Tongmai Formula group， and the rats in the model group and Yangxin Tongmai Formula group were fed with high-fat chow plus vitamin D3 by gavage plus isoproterenol hydrochloride by subcutaneous injection to construct rat models of PCHD with blood stasis syndrome. After successful modelling， rats in Yangxin Tongmai Formula group were gavaged with 18 g/（kg‧d） of Yangxin Tongmai Formula， and rats in blank group and the model group were gavaged with 4 ml/（kg‧d） of 0.9% NaCl solution， and serum samples of rats in each group were collected for DNA methylation sequencing after 3 weeks to screen for the relevant DNA methylation differentiation genes. In addition， rats with successful modelling of PCHD with blood stasis were randomly divided into model group， Yangxin Tongmai Formula with midnight-noon ebb-flow administration method group ［18 g/（kg‧d） of Yangxin Tongmai Formula was gavaged twice in the heart channel period （12：00） and pericardium channel period （20：00）］， the Yangxin Tongmai Formula control group ［18 g/（kg‧d） of Yangxin Tongmai Formula was gavaged twice at 8：00 and 18：00］ and the Atorvastatin Calcium group ［atorvastatin calcium tablets solution 1.8 mg/（kg‧d） at the same intervention time as that in Yangxin Tongmai Formula control group］， and set up a blank group of 8 rats in each group. The model group and blank group were gavaged with 0.9% NaCl solution 4 ml/（kg‧d） for the same time as the Yangxin Tongmai Formula control group. After 3 weeks of gavage， the blood lipids ［including total cholesterol （TC）， low-density lipoprotein （LDL）， high-density lipoprotein （HDL）］ levels of rats in each group were detected； the HE staining of myocardial tissues and thoracic aorta was used to observe the pathomorphological changes； the levels of serum inflammation indexes ［tumour necrosis factor alpha （TNF-alpha）， lipopolysaccharide （LPS）， and interleukin 10 （IL-10）］ were detected； immunoprecipitation-realtime fluorescence quantitative PCR was used to detect the relative expression of cardiac tissue screening differential genes. ResultsThe genes screened for differentially methylated regions were calmodulin 2 （Calm2）， calcium voltage-gated channel subunit α1s （Cacna1s）， and phospholipase Cβ1 （Plcb1）. Compared with the blank group， rats in the model group showed elevated levels of TC， LDL， TNF-α and LPS， and decreased levels of HDL and IL-10 （P＜0.05 or P＜0.01）； HE staining showed obvious swelling of myocardial fibres， accompanied by a large number of inflammatory cell infiltration， and thickening of the inner wall of the aortic vessels with internal wall damage， which was visible as a large number of lipid cholesterol crystals and obvious inflammatory cell infiltration. Compared with the model group， the TC， LDL， TNF-α and LPS contents of rats in the Yangxin Tongmai Formula with midnight-noon ebb-flow administration method group， the Yangxin Tongmai Formula control group， and the atorvastatin calcium group all reduced， and the contents of HDL and IL-10 all elevated （P＜0.05）， with the improvement of myocardial tissue damage and the reduction of inflammatory infiltration， and the improvement of the damage of the inner lining of the thoracic aorta and the reduction of lipid infiltration. Compared with Yangxin Tongmai Formula control group， LDL， TNF-α and LPS contents reduced， and IL-10 contents increased in the midnight-noon ebb-flow administration method group （P＜0.05）. Compared with the model group， the relative expression of Calm2 and Plcb1 genes decreased and the relative expression of Cacna1s gene increased in Yangxin Tongmai Formula control group and the midnight-noon ebb-flow administration method group （P＜0.05）； compared with the Yangxin Tongmai Formula control group， the relative expression of Calm2 gene decreased and the relative expression of Cacna1s gene increased in the midnight-noon ebb-flow administration method group （P＜0.05）. ConclusionThe intervention of Yangxin Tongmai Formula in the heart channel period （12：00） and pericardium channel period （20：00） was more effective in improving the blood lipid level， inhibiting inflammation， and improving myocardial tissue damage in rats of PCHD with blood stasis syndrome， and Calm2 and Cacna1s genes may be the key targets of Yangxin Tongmai Formula in intervening the blood stasis syndrome of PCHD.

Objective To observe the protective and anti-inflammatory effects of Emodin and Geniposide compatibility on the intestinal mucosal barrier function damage in rats with systemic inflammatory response syndrome(SIRS).Methods Male Sprague-Dawley(SD)rats aged 6-8 weeks were randomly divided into normal control group,model group,Emodin group,Geniposide group,Emodin and Geniposide compatibility group(compatibility group),and Ulinastatin group.The SIRS model in rats was established by abdominal injection of yeast polysaccharide.The Emodin,Geniposide,Ulinastatin,and compatibility groups received administration immediately and 12 hours after the injection of yeast polysaccharide.After 24 hours of modeling,the contents of D-lactate(D-LA),diamine oxidase(DAO),endotoxin(ET),tumor necrosis factor-α(TNF-α),interleukins(IL-1β,IL-6,IL-10)in the serum of each group were measured;The small intestine was taken for histopathological examination,and the positive protein expression levels of nuclear factor-κB p65(NF-κB p65)and Toll-like receptor 4(TLR-4)in the small intestine tissue were determined by immuno histochemistry.Results Compared with the normal control group,the levels of D-LA,DAO,ET,TNF-α,IL-1β,IL-6 and IL-10 in the serum of the model group were significantly increased[D-LA(μmol/L):99.11±11.93 vs.36.94±1.92,DAO(U/L):5 018.80±759.00 vs.2 253.23±372.40,ET(μg/L):0.36±0.04 vs.0.15±0.02,TNF-α(ng/L):66.61±20.88 vs.9.47±0.78,IL-1β(ng/L):63.73±7.64 vs.25.86±5.90,IL-6(ng/L):392.00±56.47 vs.111.17±36.22,IL-10(ng/L):41.90±8.12 vs.19.75±1.54,all P<0.05],histopathological observations showed that the small intestine mucosa was significantly swollen,the arrangement of mucosal epithelial cells was disordered,and there was cell shedding,increased infiltration of inflammatory cells in the intestinal mucosa,decreased goblet cells,loose and congested lamina propria;the positive protein expression of TLR-4 and NF-κB in the small intestine tissue was enhanced[TLR-4 positive protein expression(A value):0.59±0.08 vs.0.27±0.04,NF-κB positive protein expression(A value):0.65±0.07 vs.0.30±0.06,both P<0.05].Compared with the model group,the levels of D-LA,DAO,and ET in the serum of the Emodin group,Geniposide group,Ulinastatin group,and compatibility group were significantly decreased[D-LA(μmol/L):67.49±8.32,69.08±6.76,69.17±5.63,58.16±7.12 vs.99.11±11.93,DAO(U/L):3 659.38±563.90,3 713.29±354.70,3 575.30±444.4,3 087.01±227.50 vs.5 018.80±759.0,ET(μg/L):0.27±0.04,0.24±0.03,0.23±0.03,0.20±0.02 vs.0.36±0.04,all P<0.05],and the contents of pro-inflammatory factors TNF-α,IL-1β,and IL-6 were significantly decreased[TNF-α(ng/L):44.34±10.63,39.23±11.74,35.80±11.49,28.74±9.56 vs.66.61±20.88,IL-1β(ng/L):50.30±8.22,46.74±5.10,48.25±5.16,40.84±5.02 vs.63.73±7.64,IL-6(ng/L):299.27±50.65,263.98±37.62,281.84±63.24,216.72±38.90 vs.392.00±56.47,all P<0.05].The levels of serum anti-inflammatory factor IL-10 were significantly increased(ng/L:92.63±32.83,87.34±30.79,71.66±16.82,133.70±39.40 vs.41.90±8.12,all P<0.05).The pathological changes in the intestinal tissue of the Emodin group,Geniposide group,compatibility group,and Ulinastatin group were reduced,and the positive expressions of NF-κB p65 and TLR-4 proteins in the small intestine tissue were significantly decreased[TLR-4 positive protein expression(A value):0.49±0.03,0.47±0.08,0.36±0.08,0.42±0.06 vs.0.59±0.08,NF-κB p65 positive protein expression(A value):0.50±0.06,0.49±0.07,0.42±0.06,0.46±0.09 vs.0.65±0.07,all P<0.05].Compared with the Emodin group and Geniposide group,the serum D-LA,DAO,ET,TNF-α,IL-1β,and IL-6 in the compatibility group were significantly decreased,the serum IL-10 level was significantly increased,the pathological changes in the intestinal tissue were reduced,and the expression levels of NF-κB p65 and TLR-4 in the small intestine tissue were significantly decreased(all P<0.05).Conclusions Emodin and Geniposide can help relieve SIRS induced by yeast polysaccharide,and their effect is related to protecting the intestinal mucosal barrier and inhibiting the inflammatory response.When combined,they exhibit a synergistic effect.

Objective:To analyze clinical phenotypes and pathogenic mutations of a patient with classic tuberous sclerosis complex.Methods:Clinical data was collected from a patient with classic tuberous sclerosis complex. Next-generation sequencing was performed to screen pathogenic gene variants, and Sanger sequencing to verify the mutations. Minigene plasmids were constructed and transfected into the human renal epithelial cell line 293T, and RNA was extracted for transcriptional analysis.Results:The patient clinically presented with recurrent epileptic seizures, facial angiofibroma, periungual fibroma, pulmonary lymphangioleiomyomatosis, renal angiomyolipoma and multiple osteosclerosis. Next-generation sequencing revealed a suspected pathogenic variant in the TSC2 gene in the patient. Sanger sequencing identified a heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene in the patient, but not in his parents or 100 unrelated healthy controls. Moreover, this mutation had not been previously reported. The minigene experiment showed changed mRNA sequence of the TSC2 gene in this patient with loss of the authentic splice site in exon 4 and insertion of a 74-bp intron, which shifted the splice site 90 bp downstream (r.336delins336+16_336+90) .Conclusion:The novel heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene can lead to aberrant splicing, and may contribute to tuberous sclerosis complex in this patient.

Humans ; Autistic Disorder ; China/epidemiology* ; Cohort Studies ; Autism Spectrum Disorder/genetics* ; Asian People

Objective:To investigate the clinical effect of the anterolateral thigh perforator chimeric flap in the treatment of the wound of diabetic foot ulcer (DFU) .Methods:From January, 2018 to December, 2019, 14 cases wound of DFU of type II diabetic were treated by anterolateral thigh chimeric perforator flap. The patients were 10 males and 4 females, at 49 to 58 years old. Of the 14 patients, 10 with simple peripheral neuropathy, 4 with peripheral neuropathy complicated with vascular disease, and none with single vascular disease. With strict control of patients' blood glucose, antibiotics blended bone cement was applied or filled onto grade 2 or higher grade Wagner's DFU after debridement. In addition, the anterolateral thigh chimeric perforator flap was transferred 2 to 3 weeks later. The size of flap was 8 cm×3 cm-27 cm×7 cm. Regular followed-up were made after surgery.Results:Thirteen flaps survived in one stage after surgery. The other 1 flap had venous vascular crisis, and survived completely after active exploration. The patients were followed-up for 6-12 months. All the flaps survived well in good shape and texture. The donor and recipient areas healed well. The functional recoveries of the DF were satisfactory.Conclusion:Application of anterolateral thigh perforator chimeric flap in repair of the refractory wound of DF achieves a good clinical outcome and effectively improves the life quality of patients.

Objective    To analyze the results and diagnostic value of postoperative chylous test of pleural effusion and to verify the clinicopathological factors affecting the results of chylous test. Methods    The clinical data of 265 consecutive patients undergoing selective surgery at the Department of Thoracic Surgery, Shangjin Nanfu Hospital between May and August 2020 were retrospectively analyzed, including 106 males and 159 females with an average age of 53.0±12.2 years. According to the results of the chylous test on the operation day and postoperative first and second days, the patients were divided into two subgroups, including a positive group and a negative group, and the clinical data of the two groups were compared. Sensitivity and specificity of the chylous test were calculated. The influencing factors for chylous test were analyzed by multiple logistic regression analysis. Results    The positive rate of chylous test was 91.7%, 95.8% and 87.9% on the operation day and postoperative first and second days, respectively. There was no statistical difference in age, sex, surgical type, surgical approach, surgical site, surgical time, degree of lymph node dissection, treatment of thoracic duct, 24 hours pleural fluid drainage or 24 hours protein and fat food intake between the positive group and the negative group (P>0.05). The diagnostic sensitivity and specificity of the chylous experiment were 100.0%and 4.0%, respectively. Multiple logistic regression analysis showed that the surgical site (left/right chest) might be an influencing factor for the results of the chylous test (P=0.043, OR=0.458, 95%CI 0.216-0.974). Conclusion    The positive rate of chylous test of pleural effusion after thoracic surgery is very high. The chylous test produces a high misdiagnosis rate of chylothorax. The surgical site (left/right chest) may be an influencing factor for chylous test. The positive result of chylous test is not recommended as the direct diagnostic basis for postoperative chylothorax and guidance of the subsequent treatment.

Objective To analyze histopathological and clinical features of dermatofibroma,and to explore the relationship between them.Methods Clinical and histopathological data were collected from 150 patients with histopathologically confirmed dermatofibroma in Department of Pathology,Shanghai Skin Disease Hospital from September 2017 to August 2018,and analyzed retrospectively.Results Among the 150 patients,65 were males,and 85 were females.Their age was 42 ± 13.8 years,and the course of disease ranged from 3 months to 30 years.Some of the patients had concomitant symptoms,mainly manifesting as itching,some had spontaneous pain and mild tenderness,and 18 patients had a history of injury,insect bite or infection at lesion sites.Skin lesions mainly occurred on the extremities (107 cases,71.3％),and most were solitary (105 cases,70％).Before pathological examinations,102 cases were clinically diagnosed as dermatofibroma,16 as epidermoid cyst,13 as pigmented nevus,3 as keloid,12 as skin mass,1 as malignant melanoma,1 as xanthogranuloma,1 as prurigo nodularis,and 1 as neurofibroma.Among 169 hematoxylin and eosin (HE)-stained sections,25 (14.8％) appeared to be consistent with aneurysmal dermatofibroma,66 (39.1％)with cellular dermatofibroma,36 (21.3％) with sclerosing dermatofibroma,and 22 (13.0％)with epithelioid dermatofibroma.Coexistence of two or more subtypes could be seen in 12 sections.There were also a few new variants,such as dermatofibroma with hyperplastic sweat duct (1 case),deep dermatofibroma (3 cases),dermatofibroma with epithelioid cells intermingled with hyperplastic collagen (1 case).The duration of aneurysmal dermatofibroma varied from 7 months to 30 years,and most manifested as skin masses on the lower extremities.A relatively short course of disease was observed in patients with cellular dermatofibroma,who often visited a hospital several months after the onset,and cellular dermatofibroma was commonly observed on the extremities and frequently accompanied with itching and pain.The duration of sclerosing or atrophic dermatofibroma was usually long for years or decades,and it commonly occurred on the upper limbs without concomitant symptoms.Epithelioid dermatofibroma of varied durations had various clinical manifestations,frequently occurred on the lower limbs without concomitant symptoms.Conclusions The clinical and pathological manifestations of dermatofibroma are diverse.Different dermatofibroma lesions can share similar typical histopathological manifestations,and atypical pathological features can interfere with the diagnosis of dermatofibroma.

Oxidative stress and cardiomyocyte apoptosis are involved in the pathogenesis of doxorubicin (DOX)-induced cardiotoxicity. Matrine is well-known for its powerful anti-oxidant and anti-apoptotic capacities. Our present study aimed to investigate the effect of matrine on DOX-induced cardiotoxicity and try to unearth the underlying mechanisms. Mice were exposed with DOX to generate DOX-induced cardiotoxicity or normal saline as control. H9C2 cells were used to verify the effect of matrine . DOX injection triggered increased generation of reactive oxygen species (ROS) and excessive cardiomyocyte apoptosis, which were significantly mitigated by matrine. Mechanistically, we found that matrine ameliorated DOX-induced uncoupling protein 2 (UCP2) downregulation, and UCP2 inhibition by genipin could blunt the protective effect of matrine on DOX-induced oxidative stress and cardiomyocyte apoptosis. Besides, 5'-AMP-activated protein kinase 2 () deficiency inhibited matrine-mediated UCP2 preservation and abolished the beneficial effect of matrine in mice. Besides, we observed that matrine incubation alleviated DOX-induced H9C2 cells apoptosis and oxidative stress level activating AMPK/UCP2, which were blunted by either AMPK or UCP2 inhibition with genetic or pharmacological methods. Matrine attenuated oxidative stress and cardiomyocyte apoptosis in DOX-induced cardiotoxicity maintaining AMPK/UCP2 pathway, and it might be a promising therapeutic agent for the treatment of DOX-induced cardiotoxicity.

The atmospheric particulate matter(PM) is widely regarded as one of major environmentally and unfriendly ambient air pollution, and therein PM2.5 (diameter≤2.5 μm) is most closely related to human health.Because of its smaller diameter with longer suspension duration, PM can absorb many pathogenic microorganisms, and easily enter into the deep of airway and then deposit on the bronchus and alveoli, and it brings damage to the lung tissues and the surfactant proteins.PM can give rise to oxidative stress, inflammation response, cells and DNA damage.Now, this review focuses on the characterization and composition of PM, as well as the impact of PM2.5 on the lung, surfactant proteins and human health, which helps to call for more people to pay attention to this environmental issue in order to better mitigate and prevent the damage caused by PM.

Objective To investigate the relationship between the composition of vaginal microbiota and the course of cervical precancerous lesion.Methods A total of 64 vaginal swabs were collected from 22 healthy women, 18 CINⅠ patients and 24 CINⅡ/Ⅲ patients who visited Obstetrics and Gynecology of the Second Xiangya Hospital of Central South University during July 2014 and July 2015.The Bacterial genomic DNA was extracted and the V3 and V4 hypervariable regions of 16S rRNA were amplified and high-throughput sequenced.The abundance and composition of vaginal microbiota were analyzed by Uparse, Mothur and LefSe statistical software.Results There was no significant difference in Alpha diversity index between CINⅡ/Ⅲ group(Chao:63±32;ACE:72±38;Simpson:0.70±0.27;Shannon:0.70±0.63) and control group ( Chao:48±24;ACE:54±25;Simpson:0.71±0.27;Shannon:0.65±0.58)(W=192,P=0.11;W=189,P=0.10;W=281,P=0.72;W=241,P=0.62).The ACE(85±37) and Chao(66±25) values of CINⅠgroup were significantly different from those of the control group (ACE:54±25;Chao:48±24)(W=99,P=0.006;W=113,P=0.02).At the phylum level, 78.69%(309 020/392 722) of the vaginal microbiota in the control group was Firmicutes, 16%(62 846/392 722) was Actinobacteria.Firmicutes was reduced to 64.86%(208 422/321 318) and Actinobacteria increased to 27.71%(89 040/321 318) in CINⅠgroup.The composition of vaginal microbiotain in CINⅡ/Ⅲ group was similar to those of control group.At the genus level, the composition of vaginal microbiota were similar between CINⅡ/Ⅲ group and control group, with Lactobacillus as predominant genus[71.81%(307 658/418 424)], Gardnerella[12.91%(55 299/428 424)], others such as Prevotella, atopobium were less.In the CINⅠ group, the abundance of Lactobacillus was decreased to 56.26%(180 787/321 318), Gardnerella was increased to 19.62%(63 057/321 318), and Listeria was increased to 7.7%(24 746/321 318).The composition of vaginal microbiota in the most samples was classified as CSTⅢ and CSTⅠ, with Lactobacillus inersand and Lactobacillus crispatus were dominant respectively.There was no significant difference in the composition of vaginal microbiota between the three groups(χ2=2.72, P=0.949).LEfSe analysis showed that the abundance of bacteria in CIN group and control group were varied.At the genus level, there were significant differences in the abundance of Geobacter, Atopobium and Ureaplasma (P<0.05, P<0.05, P<0.01, respectively).At the species level, there was significant difference in the abundance of Ureaplasma urealyticum serotype 9 (P<0.01).Conclusion The diversity and the composition of vaginal microbiota were similar between CIN patients and healthy women, but the abundances of some bacteria were varied, with Ureaplasma increased in patients with CIN.