ObjectiveTo compare the differences in fungal community composition between lesional and non-lesional scalp areas in patients suffering from severe alopecia areata （AA）， and compare these with healthy scalp areas in control subjects. Additionally， to preliminarily explore the changes in scalp fungal communities in severe AA patients and their potential underlying immunological mechanisms. MethodsA total of 20 severe AA patients and 18 healthy controls were enrolled. Skin swab samples were collected from lesional and non-lesional scalp areas of severe AA patients， as well as from the normal scalp of healthy controls. The fungal internal transcribed spacer （ITS） region was amplified and analyzed using high-throughput sequencing. ResultsThe lesional scalp areas of severe AA patients exhibited higher α-diversity and species richness in fungal communities. Notably， the relative abundance of Ascomycota， along with genera such as Mycosphaerella， Aspergillus， Penicillium， and Wallemia， significantly increased in the bald regions. In contrast， Acremonium and Schizophyllum were more predominant in the non-lesional areas of severe AA patients. ConclusionDistinct region-specific differences in scalp fungal microbiota in severe AA patients suggests that fungal dysbiosis may play a potential role in the pathogenesis of alopecia areata. These findings provide new insights into the disease characteristics of severe AA from the perspective of scalp microecology.

OBJECTIVE To explore the antidepressant mechanism of Wenyang jieyu granules （WYJYG） via the NOD-like receptor thermal protein domain associated protein 3 （NLRP3）/apoptosis-associated speck-like protein containing a CARD （ASC）/Caspase-1 pathway. METHODS A rat model of depression was established by chronic unpredictable mild stress combined with single-housing for 42 consecutive days.The experiment set up blank group， model group， MCC950 （NLRP3 inflammasome inhibitor） group （10 mg/kg）， fluoxetine group （positive control，2.08 mg/kg），low-dose WYJYG（3.78 g/kg） and high-dose WYJYG group （7.56 g/kg），with 10 rats in each group. From the 22nd day of the experiment， rats in the fluoxetine group， low-dose and high-dose WYJYG groups were intragastrically administered with the corresponding drugs and intraperitoneally injected with an equal volume of normal saline. Rats in the MCC950 group were intraperitoneally injected with MCC950 at the corresponding concentration and intragastrically administered with an equal volume of distilled water. Rats in the blank group and model group were given an equal volume of distilled water by gavage and an equal volume of normal saline by intraperitoneal injection. All interventions were performed once a day for 21 consecutive days. Behavioral tests were conducted once a week. After the last administration， the contents of ASC， ionized calcium binding adaptor molecule 1 （Iba1）， interleukin-1β （IL-1β）， and IL-18 in hippocampal tissues were detected. The protein expressions of NLRP3， cluster of differentiation 68 （CD68）， Caspase-1， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein were determined， and neuronal apoptosis was observed. RESULTS After the last administration， compared with the model group， the open-field activity time was significantly prolonged （ P ＜0.05）， and the latency to feed in a novel environment was significantly shortened （ P ＜0.05） in rats of the high-dose WYJYG group. In hippocampal tissue， the contents of ASC， Iba1， IL-1β， and IL-18， as well as the protein expression levels of NLRP3， Caspase-1， and CD68， and the positive rate of neuronal apoptosis were all significantly decreased/downregulated （ P ＜0.05）. Bcl-2 protein expression was significantly upregulated （ P ＜0.05）， and the density of neuronal apoptosis-positive cells was significantly reduced （ P ＜0.05）. CONCLUSIONS WYJYG play on antidepressant role by inhibiting the NLRP3/ASC/Caspase-1 pathway， reducing microglia-mediated neuroinflammation， and inhibiting hippocampal neurons apoptosis.

Objective:To investigate the selenium levels in hair, grain, and soil samples collected from residents in typical Keshan disease (KD) endemic and non-endemic areas of Yunnan Province and Inner Mongolia Autonomous Region (referred to as Inner Mongolia), and to provide basic data for evaluating the effectiveness of KD prevention and control strategies.Methods:In 2018, one county with a history of severe KD and one non-endemic county with a similar environment were selected from each of Yunnan Province and Inner Mongolia. Within each county, one natural village was selected as the survey site. Ten soil samples were collected from each village. Ten households were stratified based on their annual per capita income levels of low, medium, and high (3, 4, 3 households, respectively). Three samples of hair and three samples of grain were collected from each household, and selenium level was measured using hydride atomic fluorescence spectrometry.Results:A total of 118 hair samples, 125 grain samples, and 39 soil samples were collected. The selenium levels in the hair of residents from KD endemic areas were significantly lower than those from non-endemic areas within the same province ( P < 0.05), and there was no statistically significant difference in selenium levels in grain and soil ( P > 0.05). The selenium level in the hair of residents in Yunnan Province [0.36 (0.31, 0.41) mg/kg] was higher than that in Inner Mongolia [0.31 (0.25, 0.35) mg/kg, Z = - 3.87, P < 0.001]. The proportion of sufficient hair selenium levels (≥0.30 mg/kg) was 86.7% (52/60) in Yunnan Province and 55.2% (32/58) in Inner Mongolia, with a statistically significant difference (χ 2 = 14.26, P < 0.001). Selenium levels in grain and soil in Yunnan Province were lower than those in Inner Mongolia ( P < 0.001). The proportion of sufficient grain selenium levels (> 0.02 mg/kg) was 5.7% (4/70) in Yunnan Province and 12.7% (7/55) in Inner Mongolia, with a significant difference (χ 2 = 11.88, P = 0.003). The selenium levels in soil in both provinces reached the sufficient level (≥0.20 mg/kg). Conclusions:The selenium levels in the population of KD endemic areas are lower than those in non-endemic regions, and the selenium deficiency among residents in Inner Mongolia is more prominent than in Yunnan Province. The selenium levels in Inner Mongolia's grain and soil are higher than those in Yunnan Province. In the future prevention and control of KD attention should be paid to monitoring the selenium nutrition level of the population in KD areas, especially in Inner Mongolia areas located in the north.

BACKGROUND:Although aromatase inhibitors significantly improve the clinical benefit of patients with hormone receptor-positive breast cancer,its associated adverse event-osteoporosis seriously affects the quality of life of patients.Huangqi Bushen Huoxue Decoction can effectively prevent the occurrence of aromatase inhibitor-induced osteoporosis,but its mechanism of action is unclear.OBJECTIVE:To investigate the effects of Huangqi Bushen Huoxue Decoction on osteoclast activity in a mouse model of osteoporosis induced by aromatase inhibitors and relevant mechanisms.METHODS:Sixty 8-week-old female C57BL/6J mice were randomly divided into sham operation group,model group,high-,medium-and low-dose Huangqi Bushen Huoxue Decoction,and positive control group,with 10 mice in each group.Bilateral ovaries were removed to establish postmenopausal animal models in all the groups except for the sham operation group.After 1 week of recovery,letrozole was injected subcutaneously to establish postmenopausal osteoporosis models via subcutaneous injection of letrozole(an aromatase inhibitor).The high-,medium-and low-dose Huangqi Bushen Huoxue Decoction groups were intragastrically given 19.24,9.62 and 4.81 g/kg/d Huangqi Bushen Huoxue Decoction(once a day),respectively.The positive control group was given alendronate 5mg/kg once a week.After 3 months of administration,Micro-CT was used to detect tibial bone mineral density and bone microstructure.Hematoxylin-eosin staining and tartrate-resistant acid phosphatase staining of the femur were performed.Immunohistochemistry was used to detect the protein expression of receptor activator of nuclear factor-κB ligand and osteoprotectin in the femur.ELISA was used to detect the serum levels of carboxyterminal cross-linked telopeptides of type I collagen and tartrate-resistant acid phosphatase 5b.RESULTS AND CONCLUSION:(1)Compared with the sham operation group,the model group showed a significant decrease in bone mineral density,sparse and fractured trabecular morphology,and a significant increase in serum levels of carboxyterminal cross-linked telopeptides of type I collagen and tartrate-resistant acid phosphatase 5b,indicating that the model of aromatase inhibitor-induced osteoporosis was successfully constructed.(2)Compared with the model group,the high-,medium-,and low-dose Huangqi Bushen Huoxue Decoction groups showed significant improvement in bone mineral density and bone microstructure,thickening and densification of trabecular morphology,significantly decreased serum levels of carboxyterminal cross-linked telopeptides of type I collagen and tartrate-resistant acid phosphatase 5b,a decrease in the number of osteoclasts and the expression of receptor activator of nuclear factor-κB ligand proteins,and an increase in the expression of osteoprotegerin.To conclude,Huangqi Bushen Huoxue Decoction may regulate the receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factor-κB/osteoprotegerin signaling pathway,inhibit osteoclast activity,improve trabecular morphology and bone microstructure,and increase bone mineral density,thus preventing the occurrence and development of aromatase inhibitor-induced osteoporosis.

AIM:To investigate the involvement of HIF-1α/VEGF regulation in its anti-angiogenesis effects using adjuvant-induced arthritis(AIA)rats.METHODS:AIA rats were orally treated by QLY ex-tract for 24 days.After sacrifice,the joints were subjected to histological examination,while the blood was used in ELISA or biochemical tests.In ad-dition,HUVEC cells were treated by QLY in vitro.MTT,wound-healing and tube-formation experi-ments were then performed.Expression of some relevant proteins in cells were investigated.RE-SULTS:Compared to the healthy controls,obvious synovial invasion and angiogenesis occurred in AIA rats.Blood levels of HIF-1α,VEGF,PDGF,and TGF-β1 were increased,while the ratio of MDA/SOD was decreased a lot.After QLY treatment,all these ab-normalities were attenuated.In vitro experiments,QLY showed notable potentials in inhibiting prolifer-ation,migration and tube-constructing abilities of HUVEC cells.Furthermore,it suppressed the ex-pression of p-MEK/MEK and p-ERK/ERK.CONCLU-SION:QLY can reduce the pathological functions of vascular endothelial cells by inhibiting HIF-1α/VEGF,and it consequently eased AIA-related abnor-mal angiogenesis in AIA rats'joints.

Objective To explore the clinical efficacy of swallowing training combined with Vitalstim neuromuscular electrical stimulation in patients with dysphagia after stroke for providing a scientific basis for improving the swallowing function.Methods A total of 100 patients with post-stroke dysphagia were enrolled and randomly divided into control group(n=50,conventional swallowing training)and observation group(n=50,conventional swallowing training+Vitalstim neuromuscular electrical stimulation),with a total treatment course of 3 weeks.The swallowing functions before and after treatment was compared between two groups.Results After treatment,observation group scored lower on the water swallow test and the Rosenbek Penetration-Aspiration Scale(P<0.05),while higher on the Functional Oral Intake Scale and the Functional Independence Measure as compared with control group(P<0.05).In addition,the score of Nutritional Risk Screening 2002(NRS2002)was higher in observation group than in control group(P<0.05).Conclusion The combination of swallowing training and Vitalstim neuromuscular electrical stimulator can effectively improve the swallowing function of stroke patients and has high clinical application value,worthy of further promotion and research.

Objective To explore the clinical efficacy of swallowing training combined with Vitalstim neuromuscular electrical stimulation in patients with dysphagia after stroke for providing a scientific basis for improving the swallowing function.Methods A total of 100 patients with post-stroke dysphagia were enrolled and randomly divided into control group(n=50,conventional swallowing training)and observation group(n=50,conventional swallowing training+Vitalstim neuromuscular electrical stimulation),with a total treatment course of 3 weeks.The swallowing functions before and after treatment was compared between two groups.Results After treatment,observation group scored lower on the water swallow test and the Rosenbek Penetration-Aspiration Scale(P<0.05),while higher on the Functional Oral Intake Scale and the Functional Independence Measure as compared with control group(P<0.05).In addition,the score of Nutritional Risk Screening 2002(NRS2002)was higher in observation group than in control group(P<0.05).Conclusion The combination of swallowing training and Vitalstim neuromuscular electrical stimulator can effectively improve the swallowing function of stroke patients and has high clinical application value,worthy of further promotion and research.

BACKGROUND:Although aromatase inhibitors significantly improve the clinical benefit of patients with hormone receptor-positive breast cancer,its associated adverse event-osteoporosis seriously affects the quality of life of patients.Huangqi Bushen Huoxue Decoction can effectively prevent the occurrence of aromatase inhibitor-induced osteoporosis,but its mechanism of action is unclear.OBJECTIVE:To investigate the effects of Huangqi Bushen Huoxue Decoction on osteoclast activity in a mouse model of osteoporosis induced by aromatase inhibitors and relevant mechanisms.METHODS:Sixty 8-week-old female C57BL/6J mice were randomly divided into sham operation group,model group,high-,medium-and low-dose Huangqi Bushen Huoxue Decoction,and positive control group,with 10 mice in each group.Bilateral ovaries were removed to establish postmenopausal animal models in all the groups except for the sham operation group.After 1 week of recovery,letrozole was injected subcutaneously to establish postmenopausal osteoporosis models via subcutaneous injection of letrozole(an aromatase inhibitor).The high-,medium-and low-dose Huangqi Bushen Huoxue Decoction groups were intragastrically given 19.24,9.62 and 4.81 g/kg/d Huangqi Bushen Huoxue Decoction(once a day),respectively.The positive control group was given alendronate 5mg/kg once a week.After 3 months of administration,Micro-CT was used to detect tibial bone mineral density and bone microstructure.Hematoxylin-eosin staining and tartrate-resistant acid phosphatase staining of the femur were performed.Immunohistochemistry was used to detect the protein expression of receptor activator of nuclear factor-κB ligand and osteoprotectin in the femur.ELISA was used to detect the serum levels of carboxyterminal cross-linked telopeptides of type I collagen and tartrate-resistant acid phosphatase 5b.RESULTS AND CONCLUSION:(1)Compared with the sham operation group,the model group showed a significant decrease in bone mineral density,sparse and fractured trabecular morphology,and a significant increase in serum levels of carboxyterminal cross-linked telopeptides of type I collagen and tartrate-resistant acid phosphatase 5b,indicating that the model of aromatase inhibitor-induced osteoporosis was successfully constructed.(2)Compared with the model group,the high-,medium-,and low-dose Huangqi Bushen Huoxue Decoction groups showed significant improvement in bone mineral density and bone microstructure,thickening and densification of trabecular morphology,significantly decreased serum levels of carboxyterminal cross-linked telopeptides of type I collagen and tartrate-resistant acid phosphatase 5b,a decrease in the number of osteoclasts and the expression of receptor activator of nuclear factor-κB ligand proteins,and an increase in the expression of osteoprotegerin.To conclude,Huangqi Bushen Huoxue Decoction may regulate the receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factor-κB/osteoprotegerin signaling pathway,inhibit osteoclast activity,improve trabecular morphology and bone microstructure,and increase bone mineral density,thus preventing the occurrence and development of aromatase inhibitor-induced osteoporosis.

BACKGROUND:At present,chemotherapeutic drugs are mainly used for the treatment of tumors,but there are problems such as drug resistance and adverse reactions.The exosome drug delivery system not only avoids the toxicity of synthetic nanoparticles,but also increases the bioavailability and biocompatibility of the drugs.It can be modified by biological,physical,and chemical methods to form a new type of nano-drug delivery platform.OBJECTIVE:To review the construction strategy of exosome drug delivery system,the application status of exosome drug delivery system in tumor diseases and the current challenges.METHODS:PubMed and CNKI were searched with"exosomal,tumor,microvesicle,extracellular vesicles,engineered,therapeutics,characterization,isolation,drug delivery,targeting,modification strategies,physics,chemistry,biology"as English search terms and"exosomes,drug delivery,tumor"as Chinese search terms.A total of 132 articles were included for in-depth induction and discussion.RESULTS AND CONCLUSION:(1)The technical methods of exosome extraction,including ultracentrifugation,filtration,and kit extraction,can efficiently isolate exosomes,but the process is complicated and time-consuming,and large-scale extraction of exosomes cannot be achieved.(2)Engineered exosomes can be divided into four categories:gene editing engineering,which improves function through genetic modification;endogenous engineering,using inflammatory factors and other pretreatment to enhance drug delivery;exogenously engineered to encapsulate drugs directly in exosomes;hybrid engineering,combining exosomes with lipid nanoparticles to form new particles.Some have entered clinical trials for cancer treatment,but most are at an early stage.In contrast,genetically engineered exosomes are considered as an important direction for future drug delivery due to their high targeting and customization potential.(3)There are still many limitations to realize the clinical transformation of engineered exosomes.At the technical level,large-scale production,purification,and drug loading efficiency are urgent to be solved.In production,high cost and batch stability affect its popularity.In terms of safety,immunogenicity and potential toxicity need to be comprehensively evaluated.Furthermore,the imperfect regulatory policies and the complexity of the approval process also constitute obstacles to its clinical translation.(4)In the future,it is necessary to promote the clinical translation process through technical innovation,cost control,safety improvement,and policy improvement.

Objective:To screen the risk factors for non-complete procedural success of transvenous lead extraction(TLE),and to establish a prediction model based on the results and evaluate its predictive efficacy.Methods:A total of 1 029 patients who underwent TLE in Peking University People's Hospital from January 2014 to December 2020 were enrolled and divided into training set(n=720)and validation set(n=309)using the random number method.There were no statistically significant differences among the variables in the training set and the validation set.The training set was divided into the complete procedural success(CPS)group(n=664)and the non-CPS group(n=56).Univariate analysis was employed to screen the relevant indicators of non-CPS,followed by binary logistic regression analysis to identify the independent risk factors of non-CPS.Subsequently,a predictive model and nomogram were constructed.The receiver operating characteristic(ROC)curve analysis was applied to evaluate the ability of the model to distinguish non-CPS from TLE patients in the training set and validation set.The Hosmer-Lemeshow goodness-of-fit test was used to assess the consistency between the predicted risk and the actual risk of the model.Results:Univariate analysis showed that the relevant variables with P<0.1 including the age at the first implantation of the lead,the number of leads extracted,the oldest dwell time of lead extracted,the presence of abandoned leads,non-manual traction for lead extracted,the number of extracted leads>3,bilateral lead implantation,and the indications for TLE.The binary logistic regression analysis revealed that the presence of abandoned leads(OR=2.252,95%CI:1.111-4.564,P=0.024),the oldest dwell time of the extracted leads(OR=1.009,95%CI:1.005-1.012,P<0.001),and the number of extracted leads>3(OR=3.177,95%CI:1.306-7.733,P=0.011)were independent risk factors for non-CPS of TLE.ROC curve analysis revealed that the area under the ROC curve(AUC)of the training set was 0.80(95%CI:0.75-0.85,P<0.001).The AUC of the validation set was 0.81(95%CI:0.72-0.90,P<0.001).The Hosmer-Lemeshow goodness-of-fit test indicated that the P values of both the training set(P=0.089)and the validation set(P=0.136)were greater than 0.05.Conclusions:The presence of abandoned leads,the oldest dwell time of lead extracted,and the number of extracted leads>3 are independent risk factors for non-CPS in patients undergoing TLE.The nomogram model based on the above factors has satisfactory predictive ability.