【Objective】 To explore the status and characteristics of voluntary blood donors in rural areas of Dali Bai Autonomous Prefecture (referred as Dali), and to provide basis for scientific and effective voluntary blood donation in rural population in regions inhabited by ethnic groups. 【Methods】 The data of rural blood donors who donated blood in Dali from 2010 to 2019 were collected, including demographic data as nationality, gender, age, educational background, as well as the blood donation frequency and blood infection screening (index) results. The above data of urban blood donors who donated blood in Dali during the same period were selected to investigate the increasing trend of blood donation rate. SPSS26.0 was used for statistical analysis of the collected data of rural and urban blood donation population. 【Results】 From 2010 to 2019,the number of blood donors in Dali increased from 13 949 to 19 479,with an increasing rate of 39.64%. The number of rural blood donors increased from 2 623 to 8 727,among which the number of ethnic minority groups increased from 1 779 to 5 059.The ratio of male to female blood donors was 70.30% (1 844/2 623) vs 29.70% (779/2 623) in 2010,56.37% (4 919/8 727) vs 43.63%(3 808/8 727) in 2019. Those with educational level of junior middle school or below were the most, accounted for 43.97%(38 443/85 836),with ethnic donors of 24.47%(23 583/85 836). The proportion of donors aged between 36 and 45 was the highest[40.73% (30 477/74 827) ], with ethnic donors of [28.56% (21 374/74 827), and the proportion of repeated blood donors was 54.87%(35 279/64 299),with ethnic donors of 49.89%(18 080/36 240) [the proportion of repeated blood donors in urban donors in the same period was 48.13% (55 677/115 675) ] (P<0.01). The unqualified rate of ALT was the highest [1.21%(1 272/105 489) ] [The unqualified rate in local donors was 1.99%(3 837/192 552) ] (P<0.01), and that of the ethnic donors was 1.51%(358/56 718). There was no significant difference in the unqualified rates of HB-sAg, anti-HCV, anti-HIV and anti-TP among urban and rural blood donors (P>0.05). 【Conclusion】 It is of great significance to explore the characteristics of blood donors in rural areas (especially regions inhabited by ethnic groups) and the reasons for disqualification, in order to scientifically carry out the recruitment of voluntary blood donors and further promote blood donation for rural residents

ObjectiveTo analyze the characters of POEMS syndrome, raise physician awareness of diagnosis and therapy,and explore the optimal treatment strategies.MethodsThe clinical features,laboratory examination and therapy of 24 patients with POEMS syndrome were analyzed,and relative literatures were reviewed. ResultsA strong predominance of male over female was found, 18 vs 6. All patients were over 40 years old,with a mean age 56.5 years old,indicating a common adult involvement.All presented with polyneuropathy,which was also the most common complaints when admitted,which reminded neurologists of underlying possibility of a rare plasma cell disorder.Organomegaly was found,including 19 cases with hepatomegaly,17 patients with splenomegaly.Endocrinal abnormalities were also found in 15 cases.18 patients were M protein positive. Skin pigmentation was recognized in 21 cases. Melphalan in combination with prednisone was applied and 100 ％ response was observed. One of the patients received peripheral blood stem cell transplantation(PBSCT)2 years ago and a durable response was observed with a continuous complete remission of polyneuropathy and the absence of M protein following 2 years post PBSCT.ConclusionPOEMS syndrome is a rare multisystem disorder, the combination of symptoms and signs is highly complex,which is easy to misdiagnose or missed diagnose.A polyneuropathy with either organomegaly,endocrinal abnormalities, skin disorder or serositis is required a further investigation of clonal M protein and bone marrow.Melphalan and prednisone probable are the optimal regimens.PBSCT provides a new choice for therapy and research of POEMS syndrome.

Objective To investigate the knowledge, attitudes and practices (KAP) towards scrub typhus control and analyse the influencing factors among residents in Longling County, Yunnan Province in 2023, so as to provide insights into management of scrub typhus in the county. Methods Two townships were randomly sampled from Longling County, Baoshan City, Yunnan Province in July 2023, and 2 to 3 administrative villages were randomly selected from each township. Then, 20 to 40 households were randomly selected from each village, and 1 to 2 villagers were surveyed in each household. Residents’ KAP towards scrub typhus control was investigated, and factor affecting residents’ KAP towards scrub typhus control were identified with independent samples t test or analysis of variance (ANOVA) and multiple linear regression analysis. Results A total of 417 questionnaires were allocated, and 410 valid questionnaires were recovered, with an effective recovery rate of 98.32%. Of all respondents, 65.85% (270/410) heard of scrub typhus but were not familiar with it, and 12.68% (52/410) and 1.95% (8/410) were familiar and very familiar with it, respectively. Residents mainly acquired scrub typhus control knowledge via their relatives and friends. The knowledge, attitude, and behavior scores of the respondents with regard to scrub typhus were (34.14 ± 13.07), (55.04 ± 6.69), and (21.40 ± 5.61), respectively, and the total score for knowledge, belief, and behavior in relation to scrub typhus prevention and control was (110.57 ± 18.80). Univariate analysis revealed that age (F = 3.12, P < 0.05), education (F = 10.77, P < 0.05), occupation (F = 4.89, P < 0.05), frequency of outdoor activities (F = 3.61, P < 0.05), and type of place of residence (t = 1.33, P < 0.05) were the factors that influenced the participants’ scores for knowledge of the prevention and control of scrub typhus. Multiple linear regression analysis revealed that literacy level (β = 10.20 to 16.71, P < 0.05) and category of residence (β = −7.23, P < 0.05) were the factors that influenced residents’ current knowledge and trust behavior in relation to scrub typhus. Conclusions Place of residence and educational level are critical factors affecting scrub typhus control among residents in Longling County, Yunnan Province. Intensified health education pertaining to scrub typhus control and appropriate behavioral interventions are required among residents living in urban areas, with a low educational level and frequent outdoor activities.

Objective To investigate the association between primary sclerosing cholangitis (PSC) and colorectal cancer (CRC) by using two-sample Mendelian randomization (TSMR). Methods The single nucleotide polymorphism (SNP) data associated with PSC and CRC were obtained from Finland Biobank and UK Biobank, respectively. A secondary data analysis was performed for all pooled data based on genome-wide association studies to select the genetic loci closely associated with PSC as instrumental variables, and TSMR was conducted by seven methods, i.e., Egger regression in Mendelian randomization, weighted median, inverse variance weighted (IVW) random effects model, maximum likelihood, linear weighted median, IVW radial method, and IVW fixed effects model. Odds ratio (OR) value was used to evaluate the causal relationship between PSC and the risk of CRC. Results There was a positive causal relationship between gene predicted PSC and CRC, and with the IVW fixed effects model as an example, genetically determined patients with PSC could increase the risk of CRC ( OR =1.002 243, 95% confidence interval: 1.001 319-1.003 167). TSMR results showed no heterogeneity ( P =0.87) or horizontal pleiotropy ( P =0.95). The three instrumental variables selected for PSC were strong instrumental variables ( F =11.86). Conclusion TSMR shows the genetic evidence for the association between PSC and the risk of CRC. Regardless of the presence or absence of inflammatory bowel disease, active enteroscopy screening among patients with PSC may help with the early identification and timely intervention of CRC.

OBJECTIVE: To explore the role of N~6-methyladenosine(m~6A) catalytic enzymes(methyltransferases and demethylases) in cadmium-induced oxidative damage in human renal epithelial cells(HK-2 cells), and to analyze the correlation between nuclear factor-erythroid 2-related factor 2(NRF2) and m~6A catalytic enzymes. METHODS: i) HK-2 cells in logarithmic growth phase were randomly divided into control group and 6 cadmium sulfate treatment groups, then treated with 0, 2, 4, 8, 16, 32 and 64 μmol/L cadmium sulfate solution for 24 hours. The cell survival rates were detected by CCK-8 assay, and the appropriate doses of cadmium sulfate were selected for subsequent experiments. ii) HK-2 cells in logarithmic growth phase were randomly divided into control group and low-, medium-, and high-dose groups, and treated with 0, 4, 8, and 16 μmol/L cadmium sulfate solution respectively for 24 hours. Subsequently, the levels of reactive oxygen species(ROS) were detected by fluorescence probe. The mRNA expression of NRF2, the m~6A methyltransferases such as methyltransferase like proteins(METTL) 3, METTL14, METTL16 and the m~6A demethylases such as fat mass and obesity associated protein(FTO), AlkB family of nonheme Fe(Ⅱ)/α-ketoglutarate(α-KG)-dependent dioxygenases 5(ALKBH5) were determined by real-time polymerase chain reaction. RESULTS: i) The survival rate of HK-2 cells was more than 60.00% and lower than that of the control group(P<0.05) after the cells were stimulated with 16 μmol/L of cadmium sulfate. Therefore, 4, 8 and 16 μmol/L of cadmium sulfate were selected as the stimulation concentrations in the follow-up experiments. ii) The relative expression of NRF2, METTL3, METTL14 and METTL16 in HK-2 cells in low-dose group increased(all P<0.05), while the levels of ROS and the relative mRNA expression of NRF2, METTL3, METTL14, METTL16 and FTO in HK-2 cells in medium and high-dose groups increased(all P<0.05) when compared with the control group. There was no significant difference in the expression of ALKBH5 mRNA among these 4 groups(P>0.05). In the correlation analysis, NRF2 mRNA expression was positively correlated with the mRNA expression of METTL3 and METTL16 [Pearson correlation coefficient(r) = 0.61 and 0.66, respectively, all P<0.05]. There was no correlation between NRF2 mRNA expression and METTL14, FTO and ALKBH5(r=0.53, 0.48, and 0.01 respectively, all P>0.05). CONCLUSION: Cadmium sulfate may increase intracellular ROS level, up-regulate NRF2 expression and activate NRF2 signaling pathway as well as enhance the expression of METTL3 and METTL16 in HK-2 cells, thus increasing intracellular oxidative damage and decreasing the cell survival rate.

OBJECTIVETo study the effect of hyperthermia on anti-invasion of Tca8113 and the expression change of matrix metalloproteinase-13 (MMP-13) and calcium-binding protein S100A4 (S100A4).

METHODSTca8113 cell pools were incubated at 43 degrees C for 0, 40, 80, 120 min, respectively, and at 37, 41, 43, 45 degrees C respectively for 80 min. The effect of high temperatures on the invasion ability of Tca8113 was measured in vitro. The slides of cells were made and incubated at 43 degrees C for 0, 40, 80, 120 min, respectively. Immunocytochemical method was employed for detecting the expression change of MMP-13 and S100A4 protein. Tca8113 cells were incubated at 43 degrees C for 0, 40, 80, 120 min respectively and at 37, 41, 43, 45 degrees C respectively for 80 min. Western blot method was conducted for detecting the expressionchange of MMP-13 and S100A4 protein.

RESULTSAs incubating time at higher temperature lasted, the proportion of the cells with invasion ability decreased. Except groups of 40 min and 80 min at 43 degrees C and 41, 43 degrees C for 80 min, the rest groups show significant statistic differences (P < 0.05). The expression intensity of MMP-13 and S100A4 proteins in Tca8113 cells would decrease as incubating time at higher temperature lasted. The content of MMP-13 and S100A4 proteins would decrease as incubating time at higher temperature lasted or incubating temperature increased. Except the groups of 40, 80 min at 43 degrees C and 41, 43 degrees C for 80 min, statistic differences were identified (P < 0.05).

CONCLUSIONThe invasion of Tca8113 could be inhibited by hyperthermia. The mechanism of this effect may be due to protein expression inhibition of MMP-13 and S100A4.

OBJECTIVE：To investigate the effects of Periplaneta americana extract on the proliferation and apoptosis of human non-small cell lung cancer A 549 cells as well as its possible mechanism. METHODS ：The dry bodies of P. americana were soaked with 90％ ethanol and eluted with gradient water-methanol by polyamide column chromatography. The 20％，30％，40％， 50％，60％，70％，80％，90％ methanol elution sites （YS-A-H）were obtained. MTT method was used to screen the active site ， and the inhibition rate of different doses of active site was detected. Flow cytometry was adopted to detect cell apoptosis ，cell cycle and mitochondrial membrane potential of cells after treated with different doses of active site. RESULTS ：Half inhibition concentrations of YS-A-H were （95.25±8.42），（129.93±7.24），（221.28±12.68），（275.39±14.87），（276.76±16.32），（31.90± 5.34），（163.15±6.97），（122.81±8.36）μg/mL，respectively. YS-F had the strongest activity. After treated with 3，9，27，81 μg/mL YS-F for 24，48，72 h，cell proliferation inhibitory rate was increased significantly at different time points ；after treated for 48，72 h，that was significantly higher than same group after treated for 24 h；after 72 h treatment ，that was significantly higher than same group after 48 h treatment （P＜0.01）. There was no significant effect of 24 h treatment of 3 μg/mL YS-F and 72 h treatment of 9 μg/mL YS-F on the percentage of cells in the late stage of necrosis，24 h treatment of 3 μg/mL YS-F on the percentage of cells in G2/M phase and 48 h treatment of 3 μg/mL YS-F on the reduction rate of mitochondrial membrane potential（P＞0.05）. The percentage of cells in the early stage of apoptosis ，the late stage of apoptosis and the early stage of necrosis ，the late stage of necrosis，as well as the percentage of cells in the Sub-G 0/G1 and S phase at each time point were significantly increased in other different doses groups ，while the percentage of cells in G 0/G1 and G 2/M phase was decreased significantly （P＜0.01）. In each dose group，the percentage of cells in the early stage of apoptosis ，the late stage of apoptosis and the early stage of necrosis ，the late stage of necrosis （except for the percentage of cells in the late stage of necrosis treated with YS-F 9 μg/mL for 72 h）and the percentage of cells in Sub-G 0/G1 phase，G2/M phase （except for YS-F 27，81 μg/mL for 48 h）after treated for 48，72 h were significantly higher than same group after 24 h of treatment ；the percentage of cells in G 0/G1 phase，S phase and G 2/M phase （except for YS-F 9 μg/mL for 48 h）after treated for 48，72 h were significantly lower than same group after 24 h of treatment （P＜0.01）；the percentage of cells in the early stage of apoptosis ，the late stage of apoptosis and the early stage of necrosis ，the late stage of necrosis （except for the percentage of cells in the late stage of apoptosis and early stage of necrosis when treated with YS-F 27 μg/mL for 72 h，the percentage of cells in the late stage of necrosis when treated with YS-F 3，9 μg/mL for 72 h were decreased significantly ）and the percentage of cells in S phase （except for YS-F 3 μg/mL for 72 h）and Sub-G 0/G1 phase after treated for 72 h were significantly higher than same group after 48 h of treatment ，while the percentage of cells in G 0/G1 and G 2/M phase were significantly lower than same group after 48 h of treatment （P＜0.01）. After treated with YS-F 9，27，81 μg/mL for 48 h，the reduction rate of cell mitochondrial membrane potential was increased significantly ；YS-F 27，81 μg/mL groups were significantly higher than YS-F 9 μg/mL group，and YS-F 81 μg/mL group was significantly higher than YS-F 27 μg/mL group. CONCLUSIONS：YS-F can inhibit the proliferation and promote the apoptosis of A 549 cells by preventing cell transformation from S phase to G 2/M phase ，and reducing mitochondrial membrane potential ，in time-dependent or dose-dependent manner.

OBJECTIVE： To study the improvement effects of Bee venom（BV） plastics on experimental cerebral thrombosis in rats. METHODS： Totally 96 SD rats were randomly divided into sham operation group （normal saline）， model group （plastics blank matrix）， Nimodipine group （positive drug， 4.00 mg/kg） and BV plastics low-dose， medium-dose， high-dose groups （1.67， 3.33， 6.67 mg/kg）， with 16 rats in each group. Rats in sham operation group and Nimodipine group were given medicine intragastrically， while rats in model group and BV plastics groups were given medicine by transdermal smearing. After 5 days of continuous administration， the experimental cerebral thrombosis model was established by ligating the right external carotid artery and pterygomandibular artery， and injecting compound thrombus inducer into the internal carotid artery. The wet mass ratio of right brain to left brain was measured to investigate the degree of brain edema on the infarcted side. The content of Evans blue （EB） in the left and right hemispheres of rats was determined by ultraviolet spectrophotometry to investigate the cerebral vascular permeability. Blood rheology and coagulation function indicators of rats were measured. The pathological changes of brain tissue in rats were observed by HE staining， and the number of survival neuron cells was counted. RESULTS： Compared with the indexes of sham operation group， the cerebral thrombosis model was established successfully. Compared with model group， the area of blue staining in the right brain （infarcted side） of rats in BV plastics groups was significantly reduced， and the right brain/left brain wet mass ratio and the content of EB in the right brain tissue were significantly reduced （P＜0.05 or P＜0.01）. The whole blood viscosity and Casson viscosity of rats in BV plastics groups， and the plasma viscosity of rats in BV plastics medium-dose and high-dose groups decreased significantly （P＜0.01）. PT and APTT of rats were prolonged significantly in BV plastics medium-dose group （P＜0.01）. The pathological changes of brain tissue in rats in BV plastics groups were significantly alleviated. The arrangement of neuron cells was more orderly， the shape and structure of cells were clear， the nucleolus was clear， the membrane was intact， and the number of survival neuron cells was significantly increased （P＜0.01）. CONCLUSIONS： BV plastics can alleviate brain edema， inhibit cerebral vascular permeability， improve hemorheology and coagulation function indicators of rats after the formation of cerebral thrombosis， and alleviate nerve cell injury after ischemia.

This study aims to investigate the role of slient mating-type information regulation 2 homolog 1(SIRT1)/tuberous sclerosis complex 2(TSC2)/mammalian target of rapamycin(mTOR) signaling pathways in the Periplaneta americana extract CⅡ-3-induced senescence of human leukemia K562 cells. K562 cells were cultured in vitro and treated with 0(control), 5, 10, 20, 40, 80, and 160 μg·mL~(-1) of P. americana extract CⅡ-3. Cell counting kit-8(CCK-8) and flow cytometry were employed to examine the proliferation and cell cycle of the K562 cells. Senescence-associated β-galactosidase stain kit(SA-β-gal) was used to detect the positive rate of senescent cells. Mitochondrial membrane potential was detected by flow cytometry. The relative mRNA level of telomerase reverse transcriptase(TERT) was determined by fluorescence quantitative PCR. The mRNA and protein levels of SIRT1, TSC2, and mTOR were determined by fluorescence quantitative PCR and Western blot, respectively. The results showed that CⅡ-3 significantly inhibited the proliferation of K562 cells and the treatment with 80 μg·mL~(-1) CⅡ-3 for 72 h had the highest inhibition rate. Therefore, 80 μg·mL~(-1) CⅡ-3 treatment for 72 h was selected as the standard for subsequent experiments. Compared with the control group, CⅡ-3 increased the proportion of cells arrested in G_0/G_1 phase, decreased the proportion of cells in S phase, increased the positive rate of SA-β-Gal staining, elevated the mitochondrial membrane potential and down-regulated the mRNA expression of TERT. Furthermore, the mRNA expression of SIRT1 and TSC2 was down-regulated, while the mRNA expression of mTOR was up-regulated. The protein expression of SIRT1 and p-TSC2 was down-regulated, while the protein expression of p-mTOR was up-regulated. The results indicated that P. americana extract CⅡ-3 induced the senescence of K562 cells via the SIRT1/mTOR signaling pathway.
Humans ; Animals ; Periplaneta ; Sirtuin 1/genetics* ; K562 Cells ; Signal Transduction ; TOR Serine-Threonine Kinases/genetics* ; RNA, Messenger ; Mammals

OBJECTIVE： To observe improvement effects of fingolimod on middle cerebral artery occlusion/reperfusion （MCAO/R） injury model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group and fingolimod low-dose， medium-dose and high-dose groups （0.5， 1， 2 mg/kg）， with 8 rats in each group. Except for sham operation group， MCAO/R injury model was induced by suture-occluded method in other groups. Administration groups were given relevant medicine intragastrically after reperfusion [1 h after reperfusion （1st day）， 22.5 h after reperfusion （2nd day）， and then every 24 h until 142.5 h of reperfusion （7th day）]. Sham operation group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 7 d. The scores of neurological deficit and balance beam test， the times of memory error [work memory error （WME）， reference memory error （RME） and total error] were recorded in each group. The contents of serum inflammatory cytokines （IL-6， IL-8， IL-10， TNF-α） were determined by ELISA， and triphenyl tetrazolium chloride staining method was used to detect the rate of cerebral infarction. RESULTS： Compared with sham operation group， neurological deficit scores （at different time points of 1st-7th day after administration）， balance beam test scores （2nd， 4th， 7th day after administration）， times of memory error （2nd， 4th， 7th day after administration）， the contents of serum inflammatory cytokines and the rate of cerebral infarction were increased significantly in model group （P＜0.05 or P＜0.01）. Compared with model group， neurological deficit scores （low-dose group at different time points of 3rd-7th day， medium-dose and high-dose groups at different time points of 2nd-7th day after administration）， balance beam test scores （low-dose group at 7th day， medium-dose group at 4th and 7th day， high-dose group at 2nd， 4th， 7th day）， RME times and total error times （low-dose group at 4th and 7th day， medium-dose group and high-dose group at 2nd， 4th， 7th day after administration）， WME times （administrations groups at 7th day after administration）， serum contents of IL-6， IL-8 and IL-10 （administrations groups）， serum contents of TNF-α （medium-dose and high-does groups） and cerebral infarction rate （medium-dose and high-dose groups） were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Intragastric administration of fingolimod can significantly reduce neurological deficit score， balance beam test score and the times of memory error in MCAO/R injury model rats， and has a protective effect on cerebral tissue and memory function. These effects may be related to the down-regulation of inflammatory cytokines such as IL-6 and TNF-α by fingolimod.