ObjectiveTo make clear exercise combined with Shenghui Tang interferes in acetylcholine receptor （M1AChR） to improve mitochondrial autophagy and enhance cognition of Alzheimer's disease （AD） model rats through the adenylate activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsForty-eight male SD rats of SPF grade were randomly divided into a blank group， a model group， a Shenghui Tang group （9.3 g·kg^-1）， an exercise group， an exercise + Shenghui Tang group （9.3 g·kg^-1）， and a rapamycin group （1.5 mg·kg^-1）. Except for the blank group， the rat model of AD was constructed by injecting amyloid beta （Aβ_1-42） into hippocampus stereotaxically. The exercise group received treadmill exercise for 4 weeks， while the Shenghui Tang group received intragastric administration for 4 weeks， and the exercise + Shenghui Tang group received treadmill exercise and intragastric administration of Shenghui Tang for 4 weeks simultaneously. After the intervention， the Morris water maze test was used to detect the learning and memory ability. Spontaneous behavior was observed in the open field test. The pathological structure of hippocampal neurons was observed by NISSl staining. The expression level of M1AChR in hippocampus was detected by immunohistochemistry （IHC）. The autophagy ultrastructure of hippocampal neurons was observed by transmission electron microscopy. The apoptosis rate was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL）. The expression of Beclin1 and microtubule-associated protein light chain 3β （LC3β） was detected by immunofluorescence （IF）. The protein expression of M1AChR， AMPK， p-AMPK， mTOR， Beclin1， LC3β， and chelate 1 （SQSTM1/p62） in hippocampus was detected by Western blot. ResultsCompared with the blank group， the model group exhibited significantly increased platform escape latency on the fifth day （P<0.01） and significantly decreased activity distance in the target quadrant and times of crossing the platform （P<0.01）. The total movement distance in the open field， the time of movement in the central area， and the average speed obviously decreased （P<0.05）. The arrangement of nerve cells in hippocampus CA1 region was dispersed， and the numbers of Nissl bodies and M1AChR positive cells significantly decreased （P<0.01）. The expression of TUNEL positive cells was significantly increased （P<0.01）. The typical autophagic lysosomal structure decreased. The protein expression of M1AChR， p-AMPK/AMPK， p-mTOR/mTOR， Beclin1， LC3Ⅱ/Ⅰ in hippocampus was significantly decreased （P<0.01）， and the protein expression of p62 was significantly increased （P<0.01）. Compared with the model group， the exercise + Shenghui Tang group exhibited obviously improved space exploration and positioning navigation ability （P<0.05， P<0.01）. The total movement distance in the open field， the time of movement in the central area， and the average speed of movement significantly increased （P<0.01）. The number of Nissl bodies significantly increased （P<0.01）. The number of M1AChR positive cells in hippocampus was significantly increased （P<0.01）. The expression of TUNEL positive cells was significantly decreased （P<0.01）. The protein expression of M1AChR， p-AMPK/AMPK， p-mTOR/mTOR， Beclin1， LC3Ⅱ/Ⅰ in hippocampus was significantly increased （P<0.01）， while the protein expression of p62 was significantly decreased （P<0.01）. Compared with the exercise + Shenghui Tang group， the Shenghui Tang group and the exercise group showed significantly increased platform escape latency on the fifth day （P<0.01） and obviously decreased activity distance in the target quadrant and times of crossing the platform （P<0.05， P<0.01）. The total movement distance in the open field， the time of movement in the central area， and the average speed of movement significantly decreased （P<0.01）. The number of Nissl bodies and the number of M1AChR positive cells significantly decreased （P<0.01）. The expression of TUNEL positive cells was obviously increased （P<0.05）. Ultrastructure of the hippocampal region showed decreased autophagy level. The protein expression of M1AChR， p-AMPK/AMPK， p-mTOR/mTOR， LC3Ⅱ/Ⅰ in the hippocampus was obviously decreased in the Shenghui Tang group （P<0.05， P<0.01）， while the protein expression of p62 was significantly increased （P<0.01）. In the exercise group， the protein expression of M1AChR， p-AMPK/AMPK， Beclin1， and LC3Ⅱ/Ⅰ was obviously decreased （P<0.05， P<0.01）， while the protein expression of p-mTOR/mTOR and p62 was significantly increased （P<0.01）. ConclusionExercise combined with traditional Chinese medicine can enhance the expression of M1AChR in the hippocampus of AD model rats， induce autophagy through the AMPK/mTOR signaling pathway， and improve the learning and memory ability of AD rats.

ObjectiveThis paper aims to investigate the effects of icariin on spermatogenesis in mice with exercise-induced fatigue and explore the underlying mechanisms. MethodsICR male mice were screened by swimming and randomly divided into normal group， model group， vitamin C group， icariin groups with low， medium， and high doses， and medium-dose icariin+N-nitro-L-arginine methyl ester （L-NAME） group， with 10 mice per group. Except for the normal group， all the other groups underwent weighted swimming training to establish an exercise-induced fatigue model. No gavage was administered during the first two weeks of the weighted training. From week three to four， the icariin groups with low， medium， and high doses received 0.03， 0.06， and 0.12 g·kg^-1 icariin via gavage， respectively. The vitamin C group received 0.2 g·kg^-1 vitamin C. The L-NAME group received 0.06 g·kg^-1 icariin and 0.01 g·kg^-1 L-NAME via intraperitoneal injection. The normal and model groups received equivalent physiological saline. After the experiment， body weight and the last exhaustive swimming time were recorded. Blood urea nitrogen （BUN）， lactate （LA）， lactate dehydrogenase （LDH）， malondialdehyde （MDA）， testicular testosterone （T）， testicular Ca²⁺/Mg²⁺-adenosine triphosphatase （ATPase） （micro-assay）， and the levels of testicular cyclic guanosine monophosphate （cGMP） were measured by using kits. Sperm CD46 levels were detected by flow cytometry. Testicular seminiferous tubules were observed via hematoxylin-eosin （HE） staining， and the testicular morphometric score （TMS） was used to evaluate the spermatogenic function. Protein expression of regucalcin （RGN， SMP30）， cGMP-dependent protein kinase 1 （PKG）， and cGMP-dependent protein kinase anchoring protein （GKAP1） was detected by Western blot. Testicular regucalcin expression was examined by immunofluorescence （IF）. The epididymal sperm quality of mice was observed under a microscope. Fluorescence-stained sections of stimulated by retinoic acid gene 8 （STRA8）， synaptonemal complex protein 3 （SCP3）， and transition protein 1（TNP1） in testicular seminiferous tubules were assessed by immunohistochemistry （IHC）. ResultsCompared with the normal group， the model group showed decreased body weight and exhaustive swimming time （P<0.01）， significantly increased fatigue markers （LA， LDH， and BUN） and lipid peroxidation product MDA （P<0.01）， reduced testicular RGN， PKG， GKAP1， testosterone， Ca²⁺/Mg²⁺-ATPase， and cGMP levels （P<0.01）， decreased sperm motility， sperm count， and TMS scores， and downregulated the expression of STRA8， SCP3， and TNP1. Compared with the model group， the icariin group with high dose exhibited increased exhaustive swimming time （P<0.01）， reduced LA， LDH， BUN， and MDA levels （P<0.01）， elevated superoxide dismutase （SOD） （P<0.01）， upregulated testicular RGN， PKG， GKAP1， testosterone， Ca²⁺/Mg²⁺-ATPase， and cGMP levels （P<0.01）， improved sperm motility， sperm count， and TMS scores， and enhanced STRA8， SCP3， and TNP1 expression. Compared with the L-NAME group， the icariin group with medium dose showed increased expression of STRA8， SCP3， and TNP1 in the testicular tissue （P<0.01） and elevated cGMP and GKAP1 levels （P<0.01）. ConclusionExercise-induced fatigue reduces the expression of RGN and cGMP/PKG/GKAP1 in mice， thereby causing abnormal spermatogenesis and impairing reproductive function in mice. Icariin ameliorates spermatogenic dysfunction in exercise-induced fatigue mice by promoting the expression of RGN and cGMP/PKG/GKAP1， thereby mitigating the damage of exercise-induced fatigue to the reproductive system.

ObjectiveThis paper aims to investigate the effects of icariin on spermatogenesis in mice with exercise-induced fatigue and explore the underlying mechanisms. MethodsICR male mice were screened by swimming and randomly divided into normal group， model group， vitamin C group， icariin groups with low， medium， and high doses， and medium-dose icariin+N-nitro-L-arginine methyl ester （L-NAME） group， with 10 mice per group. Except for the normal group， all the other groups underwent weighted swimming training to establish an exercise-induced fatigue model. No gavage was administered during the first two weeks of the weighted training. From week three to four， the icariin groups with low， medium， and high doses received 0.03， 0.06， and 0.12 g·kg^-1 icariin via gavage， respectively. The vitamin C group received 0.2 g·kg^-1 vitamin C. The L-NAME group received 0.06 g·kg^-1 icariin and 0.01 g·kg^-1 L-NAME via intraperitoneal injection. The normal and model groups received equivalent physiological saline. After the experiment， body weight and the last exhaustive swimming time were recorded. Blood urea nitrogen （BUN）， lactate （LA）， lactate dehydrogenase （LDH）， malondialdehyde （MDA）， testicular testosterone （T）， testicular Ca²⁺/Mg²⁺-adenosine triphosphatase （ATPase） （micro-assay）， and the levels of testicular cyclic guanosine monophosphate （cGMP） were measured by using kits. Sperm CD46 levels were detected by flow cytometry. Testicular seminiferous tubules were observed via hematoxylin-eosin （HE） staining， and the testicular morphometric score （TMS） was used to evaluate the spermatogenic function. Protein expression of regucalcin （RGN， SMP30）， cGMP-dependent protein kinase 1 （PKG）， and cGMP-dependent protein kinase anchoring protein （GKAP1） was detected by Western blot. Testicular regucalcin expression was examined by immunofluorescence （IF）. The epididymal sperm quality of mice was observed under a microscope. Fluorescence-stained sections of stimulated by retinoic acid gene 8 （STRA8）， synaptonemal complex protein 3 （SCP3）， and transition protein 1（TNP1） in testicular seminiferous tubules were assessed by immunohistochemistry （IHC）. ResultsCompared with the normal group， the model group showed decreased body weight and exhaustive swimming time （P<0.01）， significantly increased fatigue markers （LA， LDH， and BUN） and lipid peroxidation product MDA （P<0.01）， reduced testicular RGN， PKG， GKAP1， testosterone， Ca²⁺/Mg²⁺-ATPase， and cGMP levels （P<0.01）， decreased sperm motility， sperm count， and TMS scores， and downregulated the expression of STRA8， SCP3， and TNP1. Compared with the model group， the icariin group with high dose exhibited increased exhaustive swimming time （P<0.01）， reduced LA， LDH， BUN， and MDA levels （P<0.01）， elevated superoxide dismutase （SOD） （P<0.01）， upregulated testicular RGN， PKG， GKAP1， testosterone， Ca²⁺/Mg²⁺-ATPase， and cGMP levels （P<0.01）， improved sperm motility， sperm count， and TMS scores， and enhanced STRA8， SCP3， and TNP1 expression. Compared with the L-NAME group， the icariin group with medium dose showed increased expression of STRA8， SCP3， and TNP1 in the testicular tissue （P<0.01） and elevated cGMP and GKAP1 levels （P<0.01）. ConclusionExercise-induced fatigue reduces the expression of RGN and cGMP/PKG/GKAP1 in mice， thereby causing abnormal spermatogenesis and impairing reproductive function in mice. Icariin ameliorates spermatogenic dysfunction in exercise-induced fatigue mice by promoting the expression of RGN and cGMP/PKG/GKAP1， thereby mitigating the damage of exercise-induced fatigue to the reproductive system.

Objective To investigate the effect and mechanism of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence of human bronchial epithelial cells(HBECs).Methods CCK-8 assay was used to evaluate the cell viability in HBECs after treated with 0％,1％,2.5％,5％,7.5％and 10％CSE and/or 0,10,100,250 and 500 ng/mL rhCC16.β-galactosidase staining was used to observe the activity of the enzyme in control,CSE,and CSE+rhCC16 treated HBECs.The mRNA levels of P16 and P21 in above cell groups were detected by real-time fluorescence quantitative PCR(qPCR),and the protein levels of P16,P21,p-P53,P38,p-P38,ERK1/2 and p-ERK1/2 were detected with Western blot in control group,CSE and CSE+rhCC16 groups.Results Compared with the control group,5％CSE stimulation for 72 h or the treatment of 500 ng/mL or lower dose of rhCC16 had no significant effect on cell viability of HBECs.Stimulation 5％CSE for 72 h resulted in significantly higher positive rate to β-galactosidase staining(P＜0.05),elevated mRNA and protein levels of P16 and P21(P＜0.05),and enhanced protein levels of p-P53,p-P38 and p-ERK1/2(P＜0.05)when compared with the control group.Compared with the simple CSE stimulation,250 ng/mL 1hCC16 treatment decreased positive rate to β-galactosidase staining(P＜0.05),reduced mRNA and protein levels of P16 and P21(P＜0.05)and protein levels of p-P53,p-P38 and p-ERK1/2(P＜0.05).Conclusion rhCC16 inhibits CSE-induced cellular senescence of HBECs,which may due to its suppression in P38 MAPK and ERK1/2 signaling pathway.

Objective:To investigate the expression and prognostic value of ANO1 in oral squamous cell carcinoma(OSCC)tissues.Methods:Immunohistochemistry(IHC,n=163)and Quantitative real-time PCR(qRT-PCR,n=42)were employed to detect the expression level of ANO1 protein and mRNA in OSCC tissues and paracancerous normal tissues.The relationship between ANO1 ex-pression and clinicopathological features(n=163)and prognosis(n=93)of the patients were analyzed,and the results were compared with those in TCGA database.Results:IHC and qRT-PCR confirmed that ANO1 was highly expressed in OSCC(P＜0.05),which was consistent with the results of the TCGA database.Cox regression analysis showed that ANO1 expression was significantly correlated with T stage,lymph node metastasis and TNM stage and poor prognosis(P＜0.05).By Cox regression analysis,ANO1 overexpression(P=0.002)and differentiation degree(P=0.034)were independent prognostic factors.Conclusion:ANO1 is highly expressed in OSCC and is correlated with poor prognosis,which can be used as a novel biomarker for poor prognosis of OSCC patients.

Objective:To retrieve, appraise and integrate the best evidence of exercise rehabilitation in adult aplastic anemia patients after hematopoietic stem cell transplantation, so as to provide a reference for developing exercise rehabilitation program.Methods:All the evidences of exercise rehabilitation for aplastic anemia patients after hematopoietic stem cell transplantation were searched from the computer decision support systems, guideline websites, association official websites and databases, including guidelines, systematic reviews, evidence summaries, best practice, recommended practice, expert consensuses and randomized controlled trials. The retrieval period was from January 2010 to July 2021. Australian JBI Evidence Based Health Care Centre Evidence Grading and Evidence Recommendation Grading System was used to evaluate literature quality and evidence rating.Results:A total of 14 articles were included, and 35 evidences were summarized from 8 aspects, including exercise effectiveness, exercise principles, exercise evaluation, exercise methods, exercise intensity, exercise time, exercise site and notes.Conclusions:The summary of the best evidence for exercise rehabilitation in adult aplastic anemia patients after hematopoietic stem cell can provide a reference for clinical nursing practice. Medical and nursing staff should provide exercise guidance for patients according to the clinical practice and their own condition.

Objective To observe the clinical characteristics ofretinoblastoma (RB) in Southwest China.Methods A retrospective clinical study.From January 2010 to December 2017,66 RB patients diagnosed in Ophthalmology Department of West China Hospital of Sichuan University were included in the study.All the patients underwent ocular B-ultrasound,orbital CT or MRI examination.Ten patients underwent RetCam examination at the same time.Twenty-nine patients were diagnosed by histopathological examination,and 37 patients were diagnosed by clinical symptoms and imaging examination.According to whether the tumor invaded the orbit and optic nerve,it could be divided into extraocular stage and intraocular stage.Intraocular tumors were divided into A-E stages according to the international intraocular RB classification.Treatments were performed according to different stages.The general information,age at diagnosis,course of diseases (the time between onset symptoms and diagnosis),causes of visiting a doctor,classification,treatment methods and eyeball preservation rate were retrospectively analyzed.Results Patients all came from Southwest China (56 patients from Sichuan Province,2 patients from Yunnan Province,2 patients from Guizhou Province,and 6 patients from Tibet).The permanent residence were identified in 43 patients,including 27 patients (62.8％) from rural areas.There were 38 males (57.6％);50 unilateral tumors (75.8％) and 16 bilateral tumors (24.2％);51 firstvisiting patients (77.3％) and 15 re-visiting patients (22.7％).The average diagnostic age of first-visiting patients was 20.9 ± 14.4 months,with 23.2 ± 14.7 and 11.2 ± 7.6 months for unilateral and bilateral tumors,respectively.There were 41 patients had definite course and causes,of whom the average course was 90.6± 115.2 days.The most common cause was leucocoria in 32 patients (62.7％),followed by redness and swelling in 4 patients (9.8％),and other causes in 5 patients (12.2％).Among the 15 re-visiting patients,the average diagnostic age was 63.6± 46.8 months,the average course was 32.8 ± 45.5 months.Recurrence was occurred in 5 patients (33.3％),leucocoria in 4 patients (26.7％),postoperative complication in 3 patients (20.0％),protrusion in 2 patients (13.3％) and redness in 1 (6.7％) patient,respectively.Fifty out of 82 eyes were admitted to hospital,including 37 eyes of first-visiting patients and 13 eyes of re-visiting patients.Among 37 first-visiting eyes,there were 5 eyes (13.5％) in stage A-C,26 eyes (70.3％) in stage D-E,6 eyes (16.2％) in extraocular stage.Five eyes in stage A-C were treated with laser photocoagulation and (or) cryotherapy combined with systemic chemotherapy.Four eyes in stage D were treated with intraocular arterial chemotherapy.Nineteen eyes (51.3％) were performed with enucleation,2 eyes (5.4％) with evisceration and 7 eyes (18.9％) abandoned treatment.Among 13 re-visiting eyes,6 eyes (46.2％,with 5 eyes of recurrence) had been enucleated before,4 eyes (30.8％) were in extraocular stage and 3 eyes (23.1％) in stage D-E.Five eyes (38.5％) were treated with evisceration,4 eyes (30.8％) with enucleation,1 eye with oculoplastic surgery and 3 eyes (23.1％) abandoned treatment.The rate of eye preservation was 18.0％,29.0％ for intraocular stage and 0％ for extraocular stage,respectively.Conclusion RB patients in Southwest China have a longer course between onset symptoms and diagnosis,more advanced classification and lower rate of eye preservation.

Objective:To study the relations of human papillomavirus (HPV) infection, vaginal micro-environmental disorder with cervical lesion.Methods:A total of 1 019 women including 623 with normal cervical (NC), 303 with low-grade cervical lesion (CIN Ⅰ) and 93 with high-grade cervical lesion (CIN Ⅱ/Ⅲ) were enrolled in this study from the communities in Shanxi province, China. Case-control method was adopted, with NC as the control group and CIN as the case group. Related information was collected including demographic characteristics and relevant factors related to cervical lesions. HPV genotypes were detected by flow-through hybridization technology. Vaginal pH was detected by the pH test paper. Vaginal H 2O 2 was detected by the combined detection kit of aerobic vaginitis and bacterial vaginosis. Vaginal cleanliness was detected by smear method. Results:Data from the unconditional logistic regression analysis showed that HPV infection (CINⅠ: a OR=1.39, 95 %CI: 1.01-1.90; CINⅡ/Ⅲ: a OR=11.74, 95 %CI: 6.96-19.80), H 2O 2 (CINⅠ: a OR=2.09, 95 %CI: 1.47-2.98; CINⅡ/Ⅲ: a OR=4.12, 95 %CI: 2.01-8.43), cleanliness (CIN Ⅱ/Ⅲ: a OR=2.62, 95 %CI: 1.65-4.14), and composite indicators (CINⅠ: a OR=1.67, 95 %CI: 1.24-2.25; CINⅡ/Ⅲ: a OR=4.24, 95 %CI: 2.30-7.81) all had increased the risk of cervical lesion and the trend on the severity ( P<0.001) of cervical lesions. Additionally, we observed a synergic effect between HPV infection and vaginal micro-environmental composite indicator in CINⅡ/Ⅲ. With or without HPV infection, the ORs value of CINⅠ caused by vaginal micro-environment disorder remained close. Conclusions:Results from our study revealed that vaginal micro-environmental composite indicator could increase the risk for cervical lesion, in particular with the high-grade ones which all posed stronger risks when combined with HPV infection. However, the role of vaginal micro-environment disorder in the occurrence of CIN Ⅰ should not be ignored.

Objective : To evaluate the immune effects of inactivated H7N9 influenza vaccine. Methods : We searched several common databases（The Cochrane Library,PubMed,China Biology Medicine disc,China National Knowledge Infrastructure,etc.）for research articles about immune effects of H7N9 influenza vaccine published from the time the database built to July 10th of 2018,using H7N9 and vaccine as keywords. After screening the articles according to the inclusion and exclusion criteria,we assessed the quality of the studies and then employed seroconversion rate（SCR）as an outcome indicator to analyze the immune effects of different doses and adjuvants. Results : We recruited 5 articles on inactivated H7N9 influenza vaccine from 1 679 articles. The sample size was 2 579. The results of the meta-analysis showed that the rate difference（RD）values of SCR in each dose group after the first dose ranged from 1% to 10%,which indicated a poor protective effect;after the second dose of immunization,the RD values of SCR in the vaccines without adjuvants ranged from 13% to 19%,which was not effective enough;the RD values of SCR in the vaccines with adjuvants ranged from 62% to 69%,which met the licensing criteria for influenza vaccine;better results could be achieved when immunized with two doses of vaccines with adjuvants（ RR=1.19,95%CI：1.02-1.39）;vaccines with AS03 or MF59 at the lowest dose of 3.75 μg had the same immune effects as ones at a dose of 15 μg;vaccines with AS03（RD=89%,95%CI：85%-93%）were superior to those with MF59（RD=42%,95%CI：9%- 75%）. Conclusion Inactivated H7N9 influenza vaccines could achieve good immune effects when inoculated two doses with adjuvants,and the minimum effective dose was 3.75 μg.

Objective To compare the features of OCT angiography (OCTA) between neovascular agerelated macular degeneration (nAMD) and myopic ehoroidal neovascularization (mCNV) patients before and after intravitreal anti-VEGF treatment.Methods A prospective cohort study.Twenty-nine patients (37 eyes) with nAMD (19 males and 10 females,aged 68.20±8.76) and 31 patients (34 eyes) with mCNV (9 males and 22 females,aged 43.10± 11.80,with the mean diopter of-9.71 ± 1.20 D) from Department of Ophthalmology,West China Hospital of Sichuan University during May and December 2017 were included in this study.Ranibizumab or Conbercept (0.5 mg/0.05 ml) was intravitreally injected in all eyes.The patients were follow-up for 3-6 months.The OCTA was conducted before treatment and 1 day,1 week,1 month and 3-6 months after treatment.In order to ensure that the scanning position was the same,the tracking mode was adopted for each scanning.According to the OCTA images,the lesion area,parafoveal superficial vessel density and perfusion area were measured and analyzed contrastively between nAMD and mCNV patients.Results The mean lesion area before and 1 month after treatment in nAMD patients were 0.38± 1.87 mum2 and 0.06±0.12 mm2,while in mCNV patients,those were 0.26± 1.06 mm2 and 0.03 ± 0.05 mm2,respectively.There were statistically significant differences (Z=4.181,4.475;P＜0.001) in CNV lesion area before and 1 month after treatment between nAMD and mCNV patients.Compared with those before treatment,the absolute change (Z=1.853,P=0.064) and the percentage changes (t=2.685,P=0.010) of CNV lesion area l month after treatment in nAMD and mCNV patients show a statistical meaning.There were significantly decreases in both parafoveal superficial vessel density (F=8.997,P=0.003) and perfusion area (F=7.887,P=0.015) 3 months after treatment in nAMD patients,while decreases in parafoveal superficial vessel density (F=11.142,P=0.004) and perfusion area (F=7.662,P=0.013) could be detected 1 day after treatment in mCNV patients,before rising 1 month after treatment.Conclusions There are significantly differences in lesion area before and after the treatment of intravitreal anti-VEGF between nAMD and mCNV patients by OCTA examination.Moreover,the changes of both parafoveal superficial vessel density and perfusion area after anti-VEGF treatment are statistically different in two groups.