Aim To study the anti-itching and anti-hypersensitivity effects of Doxepin hydrochloride cream. Methods The effects of Doxepin hydrochloride cream on the pruritus of skin in mice due to 4-AP and guineapigs due to histamine phosphate were observed. The inhibitory actions of it on the increase of abdominal cavity capillary permeability in mice due to acetic acid and the angiectasis of skin in rats due to histamine were also observed. Its effects on the delayed-type hypersensitivity (DTH) of mice due to 2, 4-dinitrochlorobenzol (DNCB) and the homologous passive anaphylaxis of skin in rats were used to investigate its anti-hypersensitivity actions. Results The application of Doxepin hydrochloride cream (50, 125, 250 mg?kg -1) on the skin of mice had significant inhibitory actions on the pruritus of skin caused by 4-AP, and the application of Doxepin hydrochloride cream (10, 25, 50 mg?kg -1) on the skin of guinea pigs also had significant inhibitory actions on the pruritus of skin caused by histamine phosphate. The application of Doxepin hydrochloride cream (10,25,50 mg?kg -1 and 50,125,250 mg?kg -1) on skin inhibited the increase of skin capillary permeability in rats caused by histamine phosphate and the increase of abdominal cavity capillary permeability in mice caused by acetic acid. The application of Doxepin hydrochloride cream (50, 125, 250 mg?kg -1 and 10, 25, 50 mg?kg -1) on skin also significantly inhibited the delayed-type hypersensitivity in mice due to 2, 4-dinitrochlorobenzol and the homogeneous cutaneous anaphylaxis in rats. Conclusions Doxepin hydrochloride cream can relieve itching,inhibit the increase of capillary permeability caused by inflammatory substances. It also has significant inhibitory actions on TypeⅠ and Type Ⅳ allergic reactions.

Objective To investigate the mutations ACTN4 and SYNPO genes promoter in sporadic primary focal segmental glomerulosclerosis (FSGS) and to analyze the role of mutations in FSGS. Methods The study consisted of 82 Chinese primary FSGS, including 39 females and 43 males, ranged from 12 to 76 years old. Seventy volunteers were selected as healthy control group. Genomie DNA was extracted from peripheral blood cells of FSGS patients and hair of patients' parents by polymerase chain reaction (PCR) and direct sequencing to analyze ACTN4 and SYNPO gene promoter mutations. Mutations were matched with GenBank and TRANSFAC software database (www.ncbi.nlm.nih.gov; www.genometix.de; www.gene-regulation, corn). Dual luciferase assay system was used to analyze the promoter region mutations, based on PGL3-Basie vector, pRL-SV40 and PCI2 cell line. Hair DNA of novel mutation patients' parents was sequenced. Expression of alpha-actinin-4 and synaptopodin in patients' kidney tissue was examined by immunofluorescence. Results Three patients with 1-34C>T, 1-590delA and (1-1044delT)+ (I-797T >C) +(1-769A >G) heterozygous mutations were found in ACTN4 gene promoter respectively, and two patients with 1-24G>A and 1-851C>T heterozygous mutations in SYNPO gene promoter respectively. The same mutations were not found in the control group of 70 healthy people. Except one patient accepting her parents' 1-1044delT and 1-797T>C mutated chromosome respectively, no same mutations were found in patients' parents. Protein expression of alpha-actinin-4 and synaptopodin was reduced in mutated patients' kidneys. Except 1-1044delT group, luciferase activity in mutated groups decreased. (1-1044delT)+(1-797T>C)+(1-769A>G) mutation was associated with poor outcome and patient with these mutations progressed to end-stage renal failure. Conclusion Mutations of ACTN4 and SYNPO gene promoters affect gene transcription and protein translation, which may contribute to the onset of sporadic primary FSGS.

Objective To investigate the expression of transforming growth factor β1,transforming growth factor beta receptor(TBR)Ⅰ,TβR Ⅱ,Smad4 and C-Jun in rats with nonalcoholic fatty liver disease(NAFLD)and to find out the mechanisms of liver fibrosis in patients with NAFLD.Methods A total of 18 male SD rats were randomly divided into normal control group(n=9)and model group(n=9).The rats in control group were fed with normal diet,and those in model group were fed with fat-rich diet(consisted of 10%lard oil+2%cholesterol).An rats were sacrificed at the 20th week.The levels of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were examined by RT-PCR.The expressions of TGFβ1 and Smad4 in liver tissue were detected by immunohistochemistry.The expression of C-Jun protein was detected by Western blotting.Results The NAFLD model was successfully established.The immunohistochemistry examination revealed that TGFβ1 and Smad4 were expressed weekly in control group,but strongly expressed in model group.RT-PCR showed that A values of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were 0.46±0.12,5.z4±2.70 and 3.35±1.95,respectively,in model group,which were higher than those in control group(0.21±0.09,1.36±0.77 and 0.52±0.19,all P values＜0.01).The Western blotting results demonstrated that the expression of C-Jun protein in model group(0.93±0.41)was higher than that in control group (0.32±0.25,P=0.001).Conclusion TGFβ1/Smad4 signal pathway might be involved in the development of hepatic fibrosis in NAFLD.Blocking TGFβ1/Smad4 signal pathway will be helpful in treatment of NAFLD.

To explore the pharmacological effect of 3,4-dihydroxyacetophenone (DHAP) on the apoptosis of RAW264. 7 macrophage cells and the mechanism, RAW264. 7 macrophage cells were treated with 100 or 500 mg/L lipopolysaccharide (LPS), with or without 10-5 mol/L DHAP for 24h. Trypan blue dye exclusion assay was used to assess cell viability. Cell apoptosis was morphological studied and flow cytometric assay was used. Tumor necrosis factor-α (TNF-α) level was measured by ELISA methods. IκB protein was determined by Western blotting. Our results showed that in 100 mg/L LPS-stimulated macrophages, DHAP enhanced the cell apoptosis while in 500 mg/L LPS-stimulated macrophages, DHAP significantly inhibited the cell apoptosis. In both groups,DHAP increased the level of IκB but decreased the level of TNF-α. It is concluded that DHAP has dual effect on the apoptosis of RAW 264.7 cells treated with different concentrations of LPS. This effect may be due to the inhibition of activation of NF-κB and autocrine production of TNFα. Our study suggests that DHAP may have anti-inflammatory effect on LPS-activated macrophages.

A novel paper-based analytical device(PAD)was prepared and applied to determine the xanthine oxi-dase(XOD)inhibitory activity of Salvia miltiorrhiza extracts(SME).First,polycaprolactone was 3D printed on filter paper and heated to form hydrophobic barriers.Then the modified paper was cut according to the specific design.Necessary reagents including XOD for the colorimetric assay were immobilized on two separate pieces of paper.By simply adding phosphate buffer,the reaction was performed on the double-layer PAD.Quantitative results were obtained by analyzing the color intensity with the specialized device system(consisting of a smartphone,a detection box and sandwich plates).The 3D-printed detection box was small,with a size of 9.0 cm x 7.0 cm x 11.5 cm.Color component G performed well in terms of linearity and detection limits and thus was identified as the index.The reaction con-ditions were optimized using a definitive screening design.Moreover,a 10％glycerol solution was found to be a suitable stabilizer.When the stabilizer was added,the activity of XOD could be maintained for at least 15 days under 4℃or-20℃storage conditions.The inhibitory activity of SME was investigated and compared to that of allopurinol.The results obtained with the PAD showed agreement with those ob-tained with the microplate method.In conclusion,the proposed PAD method is simple,accurate and has a potential for point-of-care testing.It also holds promise for use in rapid quality testing of medicinal herbs,intermediate products,and preparations of traditional Chinese medicines.

Objective:To investigate the protective effect and mechanism of dexamethasone in lung ischemia/reperfusion injury (LIRI) rats.Methods:① Part one experiment: 24 Sprague-Dawley (SD) rats were divided into four groups according to the random number method ( n = 6): standard ventilation group (N group), normal saline group (NS group), LIRI group, and dexamethasone+LIRI group (DEX group). The rat model of LIRI was established by clamping the left pulmonary hilum for 1 hour and reperfusing it for 2 hours. The DEX group was given dexamethasone 3 mg/kg 5 minutes before reperfusion, and NS group was injected with normal saline. Group N did not receive any treatment. The left lung tissue of the rats in each group were taken alive 2 hours after reperfusion. The lung tissue was harvested for lung wet/dry mass ratio (W/D) measurement. Hematoxylin-eosin (HE) staining and electron microscopy was used to observe the pathological changes of lung tissue and to assess the degree of injury. Ultrastructural changes of lung tissue were observed under electron microscope. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL-1β, IL-6) in lung tissue were detected by enzyme linked immunosorbent assay (ELISA). The expressions of phosphorylated protein kinase B (p-AKT) was detected by Western Blot. ② Part two experiment: intervention with phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway inhibitor LY294002 to further explore the mechanism of dexamethasone in reducing lung injury induced by LIRI. Twenty-four SD rats were divided into four groups according to the random number method ( n = 6): N group, LIRI group, DEX group, and dexamethasone+LY294002+LIRI group (LY group). All the groups except the LY group were treated with membrane and intervention according to part one experiment. The LY group was injected with LY294002 0.3 mg/kg after injection of dexamethasone. The expressions of M1 macrophage polarization markers CD11c, CD16, and M2 macrophage polarization markers CD206, Arg1 were detected by immunohistochemistry. Results:① Part one experiment: compared with N group, the morphological and ultrastructural changes of lung tissue in the LIRI group were significantly changed, lung injury score, lung W/D ratio and TNF-α, IL-1β, IL-6 levels were significantly increased, and p-AKT expression was significantly decreased. Compared with the LIRI group, the morphological and ultrastructural changes of the lung tissue in the DEX group were significantly improved, and the lung injury score was reduced (5.00±0.89 vs. 8.83±0.75), lung W/D ratio and TNF-α, IL-1β, IL-6 levels were significantly decreased [lung W/D ratio: 6.25±0.56 vs. 8.27±0.72, TNF-α(ng/L): 93.28±16.42 vs. 205.90±25.30, IL-1β(ng/L): 130.10±10.81 vs. 209.10±19.20, IL-6 (ng/L): 195.80±21.17 vs. 310.50±20.77], p-AKT expression was significantly increased [p-AKT/AKT: (57.58±8.80)% vs. (36.62±9.25)%], and the differences were statistically significant (all P < 0.05). There was no significant difference in each index between NS group and N group. ② Part two experiment: compared with the N group, the expression of macrophage polarization markers CD11c, CD16, CD206 and Arg1 in the LIRI group were significantly increased. Compared with the LIRI group, the expressions of CD11c and CD16 in the lung tissue of the DEX group were significantly decreased, and the expressions of CD206 and Arg1 were significantly increased. The intervention of PI3K/AKT signaling pathway inhibitor LY294002 significantly blocked the effect of dexamethasone on LIRI-mediated macrophage polarization (CD11c immunohistochemical score: 7.20±0.36 vs. 5.00±0.34, CD16 immunohistochemical score: 8.20±0.48 vs. 7.40±0.64, CD206 immunohistochemical score: 5.80±0.59 vs. 7.40±0.28, Arg1 immunohistochemical score: 7.20±0.72 vs. 8.80±0.48, all P < 0.05). Conclusions:Dexamethasone pretreatment can alleviate the intrapulmonary inflammatory response and lung injury caused by LIRI in rats. The mechanism of action is related to the polarization direction of pulmonary macrophagesvia activation of the PI3K/AKT pathway by dexamethasone.

Objective:To conduct a scoping review of health behavior-related assessment tools for pregnant women with gestational diabetes mellitus (GDM).Methods:A total of 6 Chinese and English databases, including CNKI, Wanfang Database, China Biomedical Literature Database, PubMed, Web of Science Core set and Embase, were systematically searched by the terms of “gestational diabetes mellitus”，“health behavior”，“assessment”. The relevant contents of GDM health behavior-related assessment tools were retrieved for systematic analysis, and the results were normalized reported. The search period was from the establishment of the databases to February 1, 2023.Results:A total of 2 657 literatures were retrieved, and 41 literatures were finally included after screening, including 16 literatures on the development and verification of assessment tools, 2 literatures on localization of assessment tools, and 23 literatures on the application of the assessment tools. A comprehensive analysis was conducted on 18 assessment tools, including 16 original assessment tools and 2 localized assessment tools, spanning the years 1987 to 2020. The assessment content covered dietary behavior, physical activity behavior, medication adherence, blood glucose monitoring behavior, treatment adherence, self-management behavior, and health-promoting lifestyles. Among them, 7 assessment tools were validated for reliability and/or validity in pregnant women with GDM. Among the 23 studies that covered the implication of the assessment tools, the Pregnancy Physical Activity Questionnaire and Health-Promoting Lifestyle Ⅱ were the most commonly utilized tools for assessing health behaviors in pregnant women with GDM.Conclusions:There is a wide variety of assessment tools related to health behaviors in pregnant women with GDM, and the assessment content is relatively rich. However, most of the assessment tools have not been validated for their reliability and validity within the GDM population, limiting their clinical application and promotion.

Objective: To explore the relationship between screen time and prediabetes in adolescents. Methods: A cross-section study. A total of 532 adolescents aged 12~18 years in Shandong Zibo Vocational Institute were selected by stratified sampling method. The screen time was investigated by questionnaire survey. Measuring fasting fingertip blood glucose, screening for who with potentially abnormal blood glucose, and measuring their fasting venous blood glucose, which is to determine whether it is prediabetes. Results: With the increase of screen time, the risk of prediabetes increased continuously. Compared with adolescents with those who spend 0~119 minute per day screen time, the risk of prediabetes of adolescents with an average daily screen time of 120~239, 240~317, ≥318 minute increased by 0.37%, 2.63%, and 3.57%, respectively. After multi-factor adjustment, the risk of prediabetes with who take 240~317, ≥318 minute per day screen time is still higher than who with the average daily screen time of 0~119 minute, the adjusted OR was 2.502 (95% CI 1.279-4.897), 2.337(95% CI=1.189-4.594). Conclusion The results of this study show that the longer the screen time, the higher the risk of prediabetes in adolescents, and adolescents should be encouraged to reduce screen time.

OBJECTIVETo analyze the characteristics of risky behaviors among different age groups of HIV positive female sex workers, and to explore the strengthening of their management.

METHODSFrom January to June 2014, 22 814 female sex workers were investigated and tested HIV in 117 sentinel surveillance sites in Yunnan Province, and 181 were confirmed to be HIV antibody positive, who accepted questionnaire surveys. According to the age, the participants were divided into the < 35 years old age group and ≥ 35 years old age group. The demographic characteristics, knowledge about HIV/AIDS and related risk behaviors characteristics of the two groups were obtained via questionnaire surveys among 181 HIV positive female sex workers, and in-depth qualitative interviews were conducted from among 12 HIV positive sex workers.

RESULTSHIV antibody positive rate was 0.8% (181), the age of the 181 subjects were (35.83 ± 9.17) years old, 76 cases (42.0%) were < 35 years old, and 105 cases (58.0%) were ≥ 35 years old. The differences of marital status, workplace class, the last work site among two groups were statistically significant (χ(2) = 20.80, 28.32, 7.83; P < 0.001, P < 0.001, P = 0.020, respectively). Among 181 HIV, the proportion of AIDS awareness was 95.6% (173); the proportion of drug use among ≥ 35 years old age group was 51.4% (54), which was higher than that in < 35 years old age group (34.2%, 26/76) (χ(2) = 5.30, P = 0.021). 96.7% (175) received condom promotion or HIV counseling and testing in the past year. The proportion of continuing to engage in sexual services over 5 years after HIV infection was 48.5% (51/105) and the proportion of receiving antiretroviral treatment was 69.5% (73/105) in ≥ 35 years old age group, which were higher than those in the < 35 age group (30.2% (23/76), 52.6% (40/76); χ(2) = 12.26, 5.36; P = 0.002, 0.021, respectively). In-depth interviews among 12 HIV positive female sex workers found that regular clients, not consistent use of condoms were the main cause of no condom use. Economic and livelihood factors are important reasons for continuing to engage in sexual services among HIV positive sex workers.

CONCLUSIONHIV positive sex workers still have high risk behaviors including continuing to engage in commercial sexual service and no condom use after knowing their HIV infection status, and the proportion of using drugs in the ≥ 35 years old group was higher than that in < 35 years old group.

Adult ; China ; Condoms ; Female ; HIV Seropositivity ; Humans ; Marital Status ; Risk Factors ; Risk-Taking ; Safe Sex ; Sex Workers ; statistics & numerical data ; Sexual Behavior ; Substance-Related Disorders ; Surveys and Questionnaires

Using the liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine the plasma level of Lyso-GL3 in patients with Fabry disease and to analyze the clinical application of the method.

Thirty-nine patients with a genetic diagnosis of Fabry disease were included, and plasma levels of Lyso-GL3 were measured by LC-MS/MS analysis, and detailed clinical information of the patients was obtained including: α-galactosidase A activity, genetic variants, quantification of urine protein, mean arterial pressure, and estimation of glomerular filtration rate, and the differences in the levels of Lyso-GL3 in different clinical phenotypes and genotypes were statistically analyzed, as well as the association with clinical indicators.

Lyso-GL3 showed good linearity within 0.7856-400 ng/mL(r=0.9992).Further analysis of 39 Fabry disease patients diagnosed in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine showed a median Lyso-GL3 concentration of 23.6 ng/mL(4.3-92.9 ng/mL); Lyso-GL3 levels were significantly higher in patients with both the frameshift and the splicing mutations, as well as in patients with the nonsense mutations, than in patients with the missense mutations (median value 119.7 ng/mL vs. 11.9 ng/mL, P=0.006, and median value 97.0 ng/mL vs. 11.9 ng/mL, P=0.015, respectively). Whereas, association analysis revealed that Lyso-GL3 was not significantly associated with urinary protein, mean arterial pressure and estimated glomerular filtration rate.

The using of LC-MS/MS to quantify plasma Lyso-GL showed significant differences in Lyso-GL3 concentrations between classical and atypical phenotypes, suggesting that plasma Lyso-GL3 may help with clinical phenotypes. However, Lyso-GL3 levels is found to be overlapped between genotypes. No significant linear correlation was found between Lyso-GL3 and renal clinical indicators, suggesting the urgent need in finding a more accurate tool to assess renal involvement and prognosis in patients with Fabry disease.