ObjectiveTo investigate the mechanism of Ershiwuwei Guijiu pill in preventing and treating postmenopausal osteoporosis （PMOP） by activating the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway and inhibiting excessive autophagy. MethodFemale SD rats were ovariectomized and randomly divided into the sham operation group （Sham）， the operation group （OVX）， the Ershiwuwei Guijiu pill （GJ） group， and the raloxifene hydrochloride （RLX） group， with 10 rats in each group. Enzyme-linked immunosorbent assay （ELISA） and colorimetric methods were used to detect the levels of estrogen， bone metabolism markers in serum， and total superoxide dismutase （T-SOD）， malondialdehyde （MDA）， and glutathione peroxidase （GSH-Px） in tibial tissue. Flow cytometry was used to detect reactive oxygen species （ROS） levels in bone marrow mesenchymal stem cells. Masson staining was used to observe pathological changes in the proximal tibia， and micro-computed tomography （Micro-CT） was used to observe changes in tibial microstructural parameters. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the expression of autophagy-related proteins Beclin1， microtubule-associated protein 1A/1B-light chain 3 （LC3）， autophagy-related 5 （Atg5）， as well as PI3K， Akt， and mTOR in tibial tissue. ResultCompared with the Sham group， the OVX group showed a significant decrease in serum levels of estradiol （E₂） and calcium ion （Ca²⁺）， and T-SOD， GSH-Px， PI3K， Akt， and mTOR mRNA levels in bone tissue （P<0.05， P<0.01）， significantly reduced bone mineral density （BMD）， bone surface/bone volume （BS/BV）， trabecular thickness （Tb.Th）， trabecular number （Tb.N）， and trabecular connectivity （Con） in the tibia （P<0.05， P<0.01）， thinner epiphyseal growth plate， and the bone marrow cavity filled with fat vacuoles. Moreover， the levels of phosphorus （P）， MDA， ROS， and mRNA and protein expression of Beclin1， LC3， and Atg5， as well as trabecular separation （Tb.Sp） were significantly elevated （P<0.05， P<0.01）. Compared with the OVX group， the GJ and RLX groups showed significant increases in serum E₂ and Ca²⁺， and bone tissue levels of SOD， GSH-Px， and the mRNA levels of PI3K， Akt， and mTOR （P<0.05， P<0.01）， significantly increased BMD， BS/BV， Tb.Th， Tb.N， and Con in the tibia， thickened epiphyseal growth plate， and significantly reduced fat vacuoles in the bone marrow cavity （P<0.05， P<0.01）. Additionally， the levels of P， MDA， ROS， Beclin1， LC3， Atg5 mRNA and proteins， and Tb.Sp were significantly decreased （P<0.05， P<0.01）. The ratios of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR， which were significantly reduced in the OVX group （P<0.01）， were significantly increased in the GJ and RLX groups （P<0.01）. ConclusionThe Ershiwuwei Guijiu pill reduces oxidative stress and inhibits autophagy， thereby preventing and treating postmenopausal osteoporosis. Its mechanism may be related to the activation of the PI3K/Akt/mTOR signaling pathway， which inhibits autophagy.

ObjectiveTo explore the mechanism of Tibetan medicine Ershiwuwei Guijiuwan against osteoporosis in ovariectomized rats through the classical Wnt/β-catenin signaling pathway. MethodForty-eight 3-month-old SD female rats were randomized into model group （equivalent volume of normal saline）， sham operation group （equivalent volume of normal saline）， estradiol group （0.1 mg·kg^-1 estradiol valerate）， and high-， medium-， low-dose Ershiwuwei Guijiuwan groups （800， 400， 200 mg·kg^-1， respectively）. For the modeling， some adipose tissue near the ovaries was removed in the sham operation group， and ovaries were excised in other groups. Administration （ig， once a day） started two weeks after the operation and lasted 12 weeks. After sampling， based on hematoxylin and eosin （HE） staining， the morphological changes of the right femur and lumbar spine of the rats were observed. The enzyme-linked immunosorbent assay （ELISA） was performed to detect the serum levels of rat tartrate-resistant acid phosphatase 5b （TRAP5b）， collagen type Ⅰ C-terminal peptide （CTX-Ⅰ）， bone alkaline phosphatase （BALP）， amino-terminal propeptide of type Ⅰ procollagen （PINP）， and bone Gla-protein （BGP）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to examine the mRNA expression of β-catenin and Runt-related transcription factor 2 （Runx2） in femur， and Western blot to detect the protein expression of β-catenin， Runx2， and low-density lipoprotein receptor-related protein-5 （Lrp-5）. ResultCompared with the sham operation group， the model group showed disordered and sparse bone trabecula with fewer connections， decrease in serum levels of BALP， BGP， and PINP （P<0.01）， increase in levels of CTX-Ⅰ and TRAP5b （P<0.01）， reduction in mRNA expression of β-catenin and Runx2 in femoral tissue （P<0.01） and protein expression of Lrp-5， β-catenin， and Runx2 （P<0.01）. Compared with the model group， estradiol and each dose of Ershiwuwei Guijiuwan increased the volume of bone trabecula， made thebone trabecula closely arranged， reduced the loss of the trabecular meshwork， raised the serum levels of BALP and BGP （P<0.01）， and lowered TRAP5b level （P<0.01）. PINP level was significantly increased in the high-dose Ershiwuwei Guijiuwan group and estradiol group （P<0.01） and CTX-Ⅰ level was significantly decreased in the high-dose Erwenwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. The mRNA expression of β-catenin in femoral tissue and protein expression of Lrp-5 and β-catenin in estradiol group and three Ershiwuwei Guijiuwan groups were significantly increased （P<0.05， P<0.01）， and the levels of Runx2 mRNA and protein were significantly increased in the high-dose Ershiwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. ConclusionTibetan medicine Ershiwuwei Guijiuwan is effective for the osteoporosis in ovariectomized rats， which may be related to the classic Wnt/β-catenin signaling pathway. It affects bone metabolism by up-regulating the expression of osteogenesis-related genes in the signaling pathway.

Objective:To evaluate the clinical effect of applying peroneal artery perforator flap with super-draining techniques in repairing skin defects of extremities.Methods:The data of patients with limb wounds admitted to the 990th Hospital of the Joint Logistics Support Force of the People’s Liberation Army from March 2014 to August 2019 were analyzed retrospectively. All were repaired with peroneal artery perforator flaps with super-draining techniques.Cutaneous nerves and perforating vessels were anastomosed according to the routine ratio of 1∶2.The superficial vein and the recipient veinwere matched and marked according to the relative position the diameter and length.Then one to three superficial veins were anastomosed in sequence according to the vein mark number.Upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Hand Surgery Branch of the Chinese Medical Association.Patients with lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hindfoot Score System.All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association.Results:A total of 67 cases were included in this cohort. There were 54 males and 13 females, aged from 21 to 65 years, with an average of 44 years.Eight cases were anastomosed with three superficial veins, 38 cases were anastomosed with two flaps, and 21 cases were anastomosed with one flap.The donor sites were closed directly in 56 cases.Eleven cases received full-thickness skin grafting to cover the donor site.One case showed necrosis of the half flap for the arterial crisis. The wound was repaired by full-thickness skin grafting in further treatment. Three cases had hemorrhage under the free flap, which was healed secondarily after debridement. The remaining were healed well.All patients were followed for 6 to 44 months. There was no obvious swelling and blisters in the early postoperative period.The texture of the flap was the same as the surrounding area.In the later stage, the atrophy and scar were not obvious, without pigmentation. The sensation recovered quickly. The donor site healed without dysfunction.Sixteen patients with upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Chinese Medical Association Hand Surgery Branch. Twelve cases were excellent, 3 cases were good, and one case was fair. Fifty-one cases of lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hind Foot Score System. Thirty-eight cases were excellent, 11 cases were good, and two cases were fair. All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association. The two-point distance discrimination was 8-10 mm, with an average of 9.1 mm. The sensory function of the flaps reached S3+ in 48 cases and S3 in 19 cases.Conclusions:Application of super-draining technique in peroneal artery perforator flap surgery can effectively prevent blood congestion and reduce the necrosis risk of the free flap and complication rate. It can improve the survival quality of the flap and facilitate the recovery of the function of the receiving area.

Objective:To evaluate the clinical effect of applying peroneal artery perforator flap with super-draining techniques in repairing skin defects of extremities.Methods:The data of patients with limb wounds admitted to the 990th Hospital of the Joint Logistics Support Force of the People’s Liberation Army from March 2014 to August 2019 were analyzed retrospectively. All were repaired with peroneal artery perforator flaps with super-draining techniques.Cutaneous nerves and perforating vessels were anastomosed according to the routine ratio of 1∶2.The superficial vein and the recipient veinwere matched and marked according to the relative position the diameter and length.Then one to three superficial veins were anastomosed in sequence according to the vein mark number.Upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Hand Surgery Branch of the Chinese Medical Association.Patients with lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hindfoot Score System.All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association.Results:A total of 67 cases were included in this cohort. There were 54 males and 13 females, aged from 21 to 65 years, with an average of 44 years.Eight cases were anastomosed with three superficial veins, 38 cases were anastomosed with two flaps, and 21 cases were anastomosed with one flap.The donor sites were closed directly in 56 cases.Eleven cases received full-thickness skin grafting to cover the donor site.One case showed necrosis of the half flap for the arterial crisis. The wound was repaired by full-thickness skin grafting in further treatment. Three cases had hemorrhage under the free flap, which was healed secondarily after debridement. The remaining were healed well.All patients were followed for 6 to 44 months. There was no obvious swelling and blisters in the early postoperative period.The texture of the flap was the same as the surrounding area.In the later stage, the atrophy and scar were not obvious, without pigmentation. The sensation recovered quickly. The donor site healed without dysfunction.Sixteen patients with upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Chinese Medical Association Hand Surgery Branch. Twelve cases were excellent, 3 cases were good, and one case was fair. Fifty-one cases of lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hind Foot Score System. Thirty-eight cases were excellent, 11 cases were good, and two cases were fair. All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association. The two-point distance discrimination was 8-10 mm, with an average of 9.1 mm. The sensory function of the flaps reached S3+ in 48 cases and S3 in 19 cases.Conclusions:Application of super-draining technique in peroneal artery perforator flap surgery can effectively prevent blood congestion and reduce the necrosis risk of the free flap and complication rate. It can improve the survival quality of the flap and facilitate the recovery of the function of the receiving area.

Objective:To explore the mediating effect of self-directed learning ability between feedback-seeking behavior and caring ability of nursing students in school.Methods:In September 2019, convenience sampling was used to select undergraduate nursing students from grades 1 to 3 in a medical university in Liaoning Province as the research object. The Feedback Seeking Behavior Scale, Self-regulated Learning Ability Scale and the Caring Ability Inventory were used to conduct a cross-sectional survey. A total of 184 questionnaires were distributed and 172 valid questionnaires were returned.Results:The caring ability score of 172 school nursing students was (184.45±18.34) . Among nursing students in school, caring ability was positively correlated with feedback-seeking behavior and self-directed learning ability with a statistical difference ( P<0.01) , and feedback-seeking behavior was positively correlated with self-directed learning ability also with a statistical difference ( P<0.01) . The self-directed learning ability played a part of mediating role between feedback-seeking behavior and caring ability, and the difference was statistically significant ( P<0.01) , and the mediating effect was 61.18%. Conclusions:The caring ability of school nursing students is at an intermediate level and needs to be further improved. The feedback-seeking behavior of nursing students in school can directly affect the caring ability, and can also indirectly affect the caring ability through self-directed learning ability. Educators can guide and cultivate the feedback-seeking behavior and self-directed learning ability of nursing students at school, promote their self-regulation, and improve their caring abilities.

Objective: The goal of this research was to understand the demographic distribution and related factors of non-marital and non-commercial heterosexual transmission (non-commercial transmission) for HIV/AIDS (human immunodeficiency virus/acquired immunodeficiency syndrome). Methods: Data related to HIV/AIDS infected by non-marital heterosexual transmission and whose present address was in Qian Dongnan, were collected from Information System on the HIV/AIDS Prevention and Control. Information included demographic characteristics, the members of non-marital sex partners, transmission path, detection source, CD4⁺T lymphocyte level, et al. cases belong to homosexual history, injective drug use or non-classified non-marital heterosexuality transmission were excluded, totally collect HIV/AIDS 919 cases. Multivariate logistic regressions were used to analyze potential factors associated with non-marital and non-commercial heterosexual transmission. In addition, in March and June 2017, using a convenience sampling, we conducted one-to-one interviews among 10 HIV/AIDS who were infected by non-marital heterosexuality and had non-marital and non-commercial heterosexual experience in Kaili Center for Disease Control and Prevention. The content of the interview included basic information, sexual orientation, the main place of making friends and sexual behavior, attitude to commercial heterosexuality and non-martial and non-commercial heterosexuality and so on. Results: Out of the 919 cases, 645 (70.2%) were male, the proportion of non-commercial transmission was 55.06% (506). The proportion of female HIV/AIDS with non-commercial transmission was 84.7% (232), which was higher than male (42.5%(274)) (χ²=138.35, P<0.001). The proportion of Han HIV/AIDS with non-commercial transmission was 61.5% (275), which was higher than other religion (52.2%(412)) (χ²=6.32, P=0.012). The proportion of HIV/AIDS with non-commercial transmission who had 0-5 non-marital sexual partners was 58.8% (498), which was higher than who had>5 non-marital sexual partners (11.1%(8)) (χ²=61.10, P<0.001). The proportion of HIV/AIDS with non-commercial transmission who lived mobile was 72.9% (94), which was higher than who lived fixedly (52.2%(412)) (χ²=19.34, P<0.001). Qualitative interviews results revealed that the age of the respondents were 22-69. Respondents whose ages are in 22-34 were more likely to use mobile phone (4/10) and respondents whose ages are in 35-69 were less likely to look partners through party and the context of working. Conclusion The proportion of cases being infected by non-marital and non-commercial heterosexual transmission in Qian dongnan was higher than general national levels. The characteristics of sex, marriage status, migration, vocation, the members of non-marital sex partners were significant differed between commercial heterosexual transmission and non-marital and non-commercial heterosexual transmission.

Objective To find out the dietary patterns and explore the relationship between environmental factors (especially dietary patterns) and diabetes mellitus in the adults of Fujian.Methods Multi-stage sampling method were used to survey residents aged ≥18 years by questionnaire,physical examination and laboratory detection in 10 disease surveillance points in Fujian.Factor analysis was used to identify the dietary patterns,while logistic regression model was applied to analyze relationship between dietary patterns and diabetes mellitus,and classification tree model was adopted to identify the influencing factors for diabetes mellitus.Results There were four dietary patterns in the population,including meat,plant,high-quality protein,and fried food and beverages patterns.The result of logistic analysis showed that plant pattern,which has higher factor loading of fresh fruit-vegetables and cereal-tubers,was a protective factor for non-diabetes mellitus.The risk of diabetes mellitus in the population at T2 and T3 levels of factor score were 0.727 (95％CI:0.561-0.943) times and 0.736 (95％ CI:0.573-0.944) times higher,respectively,than those whose factor score was in lowest quartile.Thirteen influencing factors and eleven group at high-risk for diabetes mellitus were identified by classification tree model.The influencing factors were dyslipidemia,age,family history of diabetes,hypertension,physical activity,career,sex,sedentary time,abdominal adiposity,BMI,marital status,sleep time and high-quality protein pattern.Conclusion There is a close association between dietary patterns and diabetes mellitus.It is necessary to promote healthy and reasonable diet,strengthen the monitoring and control of blood lipids,blood pressure and body weight,and have good lifestyle for the prevention and control of diabetes mellitus.

An on-line solid phase extraction ( SPE ) coupled with HPLC-MS/MS method was developed to determine S-ammuxetine and R-ammuxetine in rat plasma. The sample preparation consisted of the following steps:A protein precipitation extraction used methanol and acetonitrile ( 50:50 , V/V ); an on-line SPE treatment to remove most matrixes in plasma;an enrichment and separation step used a C18 analytical column. S-and R-ammuxetine were determined by tandem mass spectrometry. The SPE column was a Retain PEP Javelin (10 mm × 2. 1 mm × 5 μm), while the chromatographic separation was achieved using a ZORBAX SB-C18 (50 mm × 2. 1 mm × 3. 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (40:60:0. 1, V/V/V, 0. 3 mL/min). The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 292 . 1/154 . 0 and m/z 260. 4/116. 2 were used to measure S-ammuxetine, R-ammuxetine and internal standard (propranolol). The method was linear over a concentration range from 0 . 2 to 1000 μg/L with the correlation coefficients of 0 . 9903 and 0 . 9951 . The average intra-day precision values were 1 . 2% -12 . 0% for S-ammuxetine and 0. 4%-11. 2% for R-ammuxetine, respectively. The average recoveries were 94. 2%-101. 6% for S-ammuxetine and 94. 3% -109. 4% for R-ammuxetine. Compared to the literature, the sensitivity of this method increased dramatically. The present method has been successfully applied to the preclinical rat research of ammuxetine isomers following intragastric administration.

Histone H3 lysine 4 trimethylation (H3K4me3) is well known to occur in the promoter region of genes for transcription activation.However,when investigating the H3K4me3 profiles in the mouse cerebrum and testis,we discovered that H3K4me3 also has a significant enrichment at the 3' end of actively transcribed (sense) genes,named as 3′-H3K4me3.3′-H3K4me3 is associated with ～15％ of protein-coding genes in both tissues.In addition,we examined the transcriptional initiation signals including RNA polymerase II (RNAPII)binding sites and 5′-CAGE-tag that marks transcriptional start sites.Interestingly,we found that 3′-H3K4me3 is associated with the initiation of antisense transcription.Furthermore,3′-H3K4me3 modification levels correlate positively with the antisense expression levels of the associated sense genes,implying that 3′-H3K4me3 is involved in the activation of antisense transcription.Taken together,our findings suggest that H3K4me3 may be involved in the regulation of antisense transcription that initiates from the 3′ end of sense genes.In addition,a positive correlation was also observed between the expression of antisense and the associated sense genes with 3'-H3K4me3 modification.More importantly,we observed the 3'-H3K4me3 enrichment among genes in human,fruitfly and Arabidopsis,and found that the sequences of 3'-H3K4me3-marked regions are highly conserved and essentially indistinguishable from known promoters in vertebrate.Therefore,we speculate that these 3'-H3K4me3-marked regions may serve as potential promoters for antisense transcription and 3′-H3K4me3 appear to be a universal epigenetic feature in eukaryotes.Our results provide a novel insight into the epigenetic roles of H3K4me3 and the regulatory mechanism of antisense transcription.

The global features of H3K4 and H3K27 trimethylations (H3K4me3 and H3K27me3) have been well studied in recent years,but most of these studies were performed in mammalian cell lines.In this work,wegenerated the genome-wide maps of H3K4me3 and H3K27me3 of mouse cerebrum and testis using ChlP-seq and their high-coverage transcriptomes using ribominus RNA-seq with SOLiD technology.We examined the global patterns of H3K4me3 and H3K27me3 in both tissues and found that modifications are closely-associated with tissue-specific expression,function and development.Moreover,we revealed that H3K4me3 and H3K27me3 rarely occur in silent genes,which contradicts the findings in previous studies.Finally,we observed that bivalent domains,with both H3K4me3 and H3K27me3,existed ubiquitously in both tissues and demonstrated an invariable preference for the regulation of developmentally-related genes.However,the bivalent domains tend towards a "winner-takes-all" approach to regulate the expression of associated genes.We also verified the above results in mouse ES cells.As expected,the results in ES cells are consistent with those in cerebrum and testis.In conclusion,we present two very important findings.One is that H3K4me3 and H3K27me3 rarely occur in silent genes.The other is that bivalent domains may adopt a "winner-takes-all" principle to regulate gene expression.