ObjectiveTo explore the potential mechanism underlying electroacupuncture therapy with the midnight-noon ebb-low method with hour-prescription for chronic obstructive pulmonary disease （COPD） with lung-qi deficiency syndrome from the perspective of brain and muscle basic helix-loop-helix ARNT-like 1 （BMAL1） as well as neutrophil extracellular traps （NETs）. MethodsThe experiment was conducted in two phases. For the therapeutic effectiveness observation， 24 rats were randomly allocated into control group 1， model group 1， midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group， with 6 rats per group. Rat models of COPD with lung-qi deficiency syndrome were established via cigarette smoke exposure combined with intratracheal instillation of lipopolysaccharide （LPS）. After successful modelling， rats in midnight-noon electro-acupuncture group 1 received electro-acupuncture at acupoints "Taiyuan （LU 9）"， "Feishu （BL 13）" and "Zusanli （ST 36）" during 05：00—07：00， 30 minutes per treatment once every other day for 14 consecutive days （7 sessions in total）. Rats in the conventional electro-acupuncture group received identical electroacupuncture manipulation at random daytime hours （08：00—18：00）. Pulmonary function parameters including forced expiratory volume in 0.3 second （FEV_0.3）， forced vital capacity （FVC） and FEV_0.3/FVC ratio were detected. Pulmonary histopathological changes were observed via hematoxylin-eosin （HE） staining. Plasma levels of pro-inflammatory factors interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）， as well as pulmonary reactive oxygen species （ROS） content， BMAL1 protein expression and the levels of neutrophil extracellular traps （NETs） biomarkers such as myeloperoxidase （MPO）， neutrophil elastase （NE）， and citrullinated histone H3 （CitH3） in lung tissue were determined. For the mechanistic verification phase， 36 rats were divided into control group 2， model group 2， midnight-noon electro-acupuncture group 2， overexpression empty group， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group， with 6 rats in each group. At the 5th week of model construction， rats in the overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group received intratracheal instillation of 100 μl adenoviral suspension carrying overexpressed Bmal1 gene. Rats in the overexpression empty group were injected with an equal titer and equal volume of blank adenovirus without the Bmal1 coding sequence into the lung at the identical time point. Midnight-noon electro-acupuncture group 2 and overexpression with midnight-noon electro-acupuncture group were consistent with the aforementioned protocol. Finally， pulmonary levels of MPO， NE and CitH3 were measured in all groups. ResultsCompared with the control group 1， the model group 1 exhibited decreased FEV_0.3， FVC and FEV_0.3/FVC ratio， elevated plasma IL-1β， TNF-α and pulmonary ROS levels， downregulated lung BMAL1 protein expression， and increased contents of MPO， NE and CitH3 （all P＜0.05）； typical NETs-like structures with sparse granular substances attached to fibrous networks were observed under scanning electron microscopy； HE staining revealed damaged alveolar architecture accompanied by inflammatory cell infiltration. Compared with model group 1， both the midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group achieved obvious improvements in all above indicators， with superior therapeutic effects in midnight-noon electro-acupuncture group 1 （P＜0.05）. Consistently， midnight-noon electro-acupuncture group 1 showed more prominent alleviation of NETs-like structure formation and lung pathological injury compared to the conventional electro-acupuncture group. Mechanistic validation results demonstrated that compared with model group 2， the average optical density of MPO as well as the protein levels of MPO， NE and CitH3 were markedly reduced in midnight-noon electro-acupuncture group 2， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group （P＜0.05）； furthermore， these parameters were significantly lower in the overexpression with midnight-noon electro-acupuncture group than in midnight-noon electro-acupuncture group 2 （P＜0.05）. ConclusionElectro-acupuncture based on the midnight-noon ebb-low method with hour-prescription may alleviate pulmonary inflammation in COPD by upregulating the expression of clock protein BMAL1 and inhibiting excessive accumulation of NETs in lung tissue.

AIM:To explore the mechanism of Chonghe paste in regulating PTEN-induced kinase 1(PINK1)/parkin pathway-mediated mitophagy on diabetic ulcer wound healing.METHODS:Thirty-six rats were used to construct a diabetic ulcer model by high-sugar and high-fat feed,intraperitoneal injection of streptozotocin and skin defect method,and were randomly divided into model group,Chonghe paste group and growth factor group,with 12 rats in each group.Another 12 rats were taken to construct normal wound model as normal group.The rats in normal group and model group were treated with normal saline for debridement,while those in Chonghe paste group and growth factor group were treated with Chonghe paste and growth factors for wound coating,respectively.The changes in fasting blood glucose and wound healing were observed and recorded,and samples were collected on the 7th and 14th days after treatment.The expression levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),P62 and collagen type Ⅲ(COL3)in granulation tissues were detected by immunohistochemistry.The mitochondrial ultrastructure was observed under an electron microscope,and the number of autophagosomes was recorded.The expression of PINK1,parkin and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)in wound tissues was detected by immunofluorescence,Western blot and RT-qPCR.RE-SULTS:Compared with normal group,the blood glucose levels of the rats in model group,Chonghe paste group and growth factor group were higher than 16.67 mmol/L on the 7th,14th and 28th days,indicating successful modeling of dia-betes.Compared with normal group,the rats in model group had a lower wound healing rate,the mean absorbance of IL-6,TNF-α and P62 was increased,the mean absorbance of COL3 was decreased,the number of autophagosomes per unit area was increased,the mean absorbance of PINK1,parkin and LC3-Ⅱ was decreased,and the mRNA and protein expres-sion levels of PINK1,parkin and LC3-Ⅱ were decreased.Compared with model group,the wound healing rate of the rats in Chonghe paste group and growth factor group was increased,the mean absorbance of IL-6,TNF-α and P62 was de-creased,the mean absorbance of COL3 was increased,the number of autophagosomes per unit area was decreased,the mean absorbance of PINK1,parkin and LC3-Ⅱ was increased,and the mRNA and protein expression levels of PINK1,parkin and LC3-Ⅱ were elevated.CONCLUSIONS:Chonghe paste can alleviate local inflammatory response in diabetic ulcer wounds,reduce the levels of inflammatory factors IL-6 and TNF-α,promote COL3 expression,activate the PINK1/parkin signaling pathway,and promote mitophagy in wounds,thus accelerating wound healing.

AIM:To explore the mechanism of Chonghe paste in regulating PTEN-induced kinase 1(PINK1)/parkin pathway-mediated mitophagy on diabetic ulcer wound healing.METHODS:Thirty-six rats were used to construct a diabetic ulcer model by high-sugar and high-fat feed,intraperitoneal injection of streptozotocin and skin defect method,and were randomly divided into model group,Chonghe paste group and growth factor group,with 12 rats in each group.Another 12 rats were taken to construct normal wound model as normal group.The rats in normal group and model group were treated with normal saline for debridement,while those in Chonghe paste group and growth factor group were treated with Chonghe paste and growth factors for wound coating,respectively.The changes in fasting blood glucose and wound healing were observed and recorded,and samples were collected on the 7th and 14th days after treatment.The expression levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),P62 and collagen type Ⅲ(COL3)in granulation tissues were detected by immunohistochemistry.The mitochondrial ultrastructure was observed under an electron microscope,and the number of autophagosomes was recorded.The expression of PINK1,parkin and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)in wound tissues was detected by immunofluorescence,Western blot and RT-qPCR.RE-SULTS:Compared with normal group,the blood glucose levels of the rats in model group,Chonghe paste group and growth factor group were higher than 16.67 mmol/L on the 7th,14th and 28th days,indicating successful modeling of dia-betes.Compared with normal group,the rats in model group had a lower wound healing rate,the mean absorbance of IL-6,TNF-α and P62 was increased,the mean absorbance of COL3 was decreased,the number of autophagosomes per unit area was increased,the mean absorbance of PINK1,parkin and LC3-Ⅱ was decreased,and the mRNA and protein expres-sion levels of PINK1,parkin and LC3-Ⅱ were decreased.Compared with model group,the wound healing rate of the rats in Chonghe paste group and growth factor group was increased,the mean absorbance of IL-6,TNF-α and P62 was de-creased,the mean absorbance of COL3 was increased,the number of autophagosomes per unit area was decreased,the mean absorbance of PINK1,parkin and LC3-Ⅱ was increased,and the mRNA and protein expression levels of PINK1,parkin and LC3-Ⅱ were elevated.CONCLUSIONS:Chonghe paste can alleviate local inflammatory response in diabetic ulcer wounds,reduce the levels of inflammatory factors IL-6 and TNF-α,promote COL3 expression,activate the PINK1/parkin signaling pathway,and promote mitophagy in wounds,thus accelerating wound healing.

Breast cancer is the most common malignancy in females,often requiring mastectomy for treatment.While mastectomy im-proves survival time,it imposes physical and psychological challenges.Thus,there is a requirement for breast reconstruction that is not only aesthetically effective but also natural to the touch.Nerve grafting has emerged as a promising technique for restoring sensory function in these patients,and this review explores how nerve grafts are used in breast reconstruction.Multiple nerve grafting techniques in breast re-construction and some of the problems associated with them were identified,which provides significant guidance for future studies.

Objective To construct a Type 1 diabetes model in miniature pigs and explore postoperative care strategies for effectively prolonging the survival time of the model pigs.Methods Seven Banna miniature pigs were selected for pancreatectomy.Glucose,vitamins,and antibiotics were administered for 3-5 days after surgery to aid recovery.Blood glucose and urine glucose levels were measured twice a day in the morning and evening to adjust insulin supplementation accordingly.The model pigs were observed daily and records were kept,including orexis,psychosis,weakness,skin ulcer,and feces and urine.Body weight was measured weekly until the death of the model animals.Based on the model pigs'condition,glucose injection and Ringer's lactate solution were administered to supplement nutrition and correct electrolyte imbalances.Results All seven Banna miniature pigs showed typical symptoms of diabetes:random blood glucose levels higher than 11.1 mmol/L after pancreatectomy,far exceeding the average blood glucose level of 6.0 mmol/L in normal pigs;positive urine glucose;and progressive weight loss.These features indicated the successful construction of Type 1 diabetes model.Additionally,Type 1 diabetic pigs that survived more than 8 weeks showed progressive hair loss and skin ulceration.Euthanasia was performed on model pigs when they were unable to stand or even eat independently,and pathological examination and HE staining were conducted on tissues collected from affected organs such as the liver,kidneys,and skin.Pathological sections revealed liver congestion,massive glycogen accumulation,ballooning degeneration of hepatocytes,and progressive liver fibrosis,along with glomerular congestion,vacuolar degeneration in renal tubular epithelial cells,proteinuria,dermal congestion,thinning of vascular walls,and varying degrees of parakeratosis and dyskeratosis in the liver,kidneys,and skin tissues due to prolonged hyperglycemia.The average survival time of the constructed Banna miniature pig diabetes model was 44 d,with a maximum survival time of 121 d.Conclusion Type 1 diabetes model can be constructed successfully in Banna miniature pigs through pancreatectomy.With meticulous postoperative care,a long-term Type 1 diabetes model with significant complications can be achieved,providing a stable large-animal model for Type 1 diabetes treatment strategies.

Objective To investigate the mechanism of Chonghe soft extract on ulcer wound healing in diabetic rats through protein kinase B(Akt)/mammalian Sirolimus target protein(mTOR)-mediated nucleotides binding oligomeric acid domain-like receptor protein 3(NLRP3)inflammasome inactivation.Methods Thirty six SD rats with diabetic ulcer,which were established by feeding with high glucose and high fat diet and injecting intraperitoneally with streptozocin(STZ)combined with skin defect,were randomly divided into model group,Chonghe soft extract group and growth factor group,with twelve rats in each group.Another twelve SD rats were injected an equal dose of citric acid-sodium citrate buffer solution and used as blank group.The blank group and the model group were not received drug intervention,but the Chonghe soft extract group and the growth factor group were externally applied Chonghe soft extract and growth factor gel,respectively.The wound healing of each group was observed and recorded.After 7 days and 14 days of treatment,the histopathology of wound were observed by HE staining and the number of fibroblasts were counted.The levels of IL-1β,IL-18 and TNF-α in serum were detected by ELISA.The expression of autophagy-related protein Beclin-1 and LC3Ⅱ in granulation tissue was detected by immunohistochemistry.The expression of NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),Caspase1,Pro-Caspase1 and Akt/mTOR autophagy pathway-related proteins Akt,p-Akt,mTOR and p-mTOR were detected by Western Blot.Results Compared with the blank control group,the pathological wound repair of the model group was delayed on the 7th day and 14th day,the number of fibroblasts per unit area was decreased(P<0.01).The levels of IL-1β,IL-18 and TNF-α were increased(P<0.01).The expression levels of ASC,Pro-Caspase1,Caspase1,and NLRP3 were increased in the wound tissues(P<0.01),while the expression levels of Beclin-1,LC3-Ⅱ,mTOR,p-mTOR,Akt and p-Akt were decreased in the wound tissues(P<0.01).Compared with the model group,the pathological injury in Chonghe soft extract group and growth factor group was significantly improved on the 7th day and 14th day.The number of fibroblasts per unit area was significantly increased(P<0.01).The levels of IL-1β,IL-18 and TNF-α were significantly decreased(P<0.01).The expression levels of ASC,Pro-Caspase1,Caspase1,and NLRP3 in the wound tissues were decreased(P<0.01),while the expression levels of Beclin-1,LC3-Ⅱ,mTOR,p-mTOR,Akt and p-Akt were increased(P<0.01,P<0.05).Conclusion Chonghe soft extract can reduce inflammatory reaction,promote the generation of fibro,regulate the Akt/mTOR-mediated NLRP3 inflammasome inactivation,improve the level of autophagy in wound,and promote ulcer wound healing in diabetic rats.

This study was performed to investigate the features of HBV-specific CD8+T cell reactivity in patients with chronic hepatitis B(CHB),HBV-induced liver cirrhosis(LC)or hepatocellular carcinoma(HCC).A total of 124 CHB patients,36 LC patients,and 114 HCC patients were enrolled in this study.The reactive HBV-specific CD8+T cells in peripheral blood were enumerated using an innovative ELISPOT system.In addition,19 CHB patients and 20 HCC patients were longitudinally monitored with an interval of 3-5 months.Data showed that the numbers of reactive HBV-specific CD8+T cells in CHB group were not significantly different from that in LC group,but obviously lower than that in HCC group(P=0.009 9),especially HBsAg-,HBpol-and HBe/cAg-specific CD8+T cells.In CHB group,the patients with normal ALT level,AST level,or low HBV-DNA load showed significantly more reactive HBV-specific CD8+T cells than the patients with abnormal ALT level,abnormal AST level,or high HBV-DNA load.Furthermore,the duration of NUCs treatment had an impact on the HBV-specific CD8+T cell reactivity in CHB patients,while different NUCs at the same treatment duration did not bring different reactivity of HBV-specific T cells.In LC group,the HBeAg-positive patients presented much more reactive HBV-specific CD8+T cells than the HBeAg-negative patients did.In HCC group,the numbers of reactive HBV-specific CD8+T cells in the patients with normal AFP level or normal DCP level were significantly higher than that in the patients with abnormal AFP level or abnormal DCP level.Longitudinal monitoring results showed that HBV-specific CD8+T cell reactivity displayed a slow upward trend in the CHB patients undergoing NUCs treatment,and an obvious increasing in the HCC patients undergoing combined treatment of targeted drugs and immunotherapy.Taken together,the features of HBV-specific CD8+T cell reactivity are distinct among the CHB,LC and HCC patients,and are influenced by virological indicators,tumor markers and treatment regimens.Therefore,more attention should be paid to the changes of HBV-specific CD8+T cell reactivity during clinical treatment.

Breast cancer is the most common malignancy in females,often requiring mastectomy for treatment.While mastectomy im-proves survival time,it imposes physical and psychological challenges.Thus,there is a requirement for breast reconstruction that is not only aesthetically effective but also natural to the touch.Nerve grafting has emerged as a promising technique for restoring sensory function in these patients,and this review explores how nerve grafts are used in breast reconstruction.Multiple nerve grafting techniques in breast re-construction and some of the problems associated with them were identified,which provides significant guidance for future studies.

Under the organization of Renji Hospital, Shanghai Jiao Tong University School of Medicine, a specialized disease database of inflammatory bowel disease (IBD) cohort was deployed, and a brief introduction of the database was made in this article. The IBD data set was established by referring to domestic and foreign standards. Through data extraction, cleaning, normalization and other information processing technologies, data from multi‑source heterogeneous platform were arranged to form a specialized major disease database of IBD cohort and the efficiency and quality of data collection in clinical practice, teaching and scientific research were guaranteed. The display and personalized export capacities of the database can promote the researches on IBD and assist the clinical decision‑making. It provides not only efficient, comprehensive and reliable research‑level data support for scientific research, but also a precise guidance for diagnosis and treatment of the disease. Furthermore, it can excavate the potential clinical principles based on medical big data.

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