[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34⁺ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.

BACKGROUND: Cardiovascular disease (CVD) is the predominant cause of mortality in China. However, the mechanisms linking sarcopenia to CVD remain poorly understood, particularly in normal-weight populations. Individuals with the absence of overweight or obesity may tend to experience missed opportunities for timely intervention. This study aimed to investigate the longitudinal association between sarcopenia and incidence of new-onset CVD in a normal-weight population, and to examine the mediating effect of functional limitation in this relationship. METHODS: We conducted a closed-cohort analysis using a nationwide sample of 4,147 middle-aged and older adults with normal weight in China. We performed Cox proportional hazards regression analysis to explore the associations of baseline sarcopenia with incident CVD. The difference method was applied to estimate the mediation proportion of functional limitation in this association. RESULTS: Over a mean follow-up period of 7.62 years, CVD occurred in 835 participants. In the multivariable-adjusted Cox model, individuals with sarcopenia exhibited a significantly higher likelihood of developing incident CVD compared to those without sarcopenia (adjusted hazard ratio [aHR] = 1.45, 95% confidence interval [CI]: 1.21-1.73, P < 0.001). Similar associations were observed for the incidence of heart disease and stroke. Functional limitation accounted for approximately 15.0% of the total effect of sarcopenia on incident CVD (P < 0.001). CONCLUSIONS Sarcopenia exerts both direct and indirect effects on incident CVD among middle-aged and older adults who are normal weight, with functional limitation serving as a significant mediator. Interventions targeting both sarcopenia and functional limitation may offer a promising strategy for enhancing cardiovascular health in this population.
Humans ; Sarcopenia/complications* ; China/epidemiology* ; Male ; Female ; Middle Aged ; Cardiovascular Diseases/etiology* ; Aged ; Incidence ; Cohort Studies ; Proportional Hazards Models ; Risk Factors ; Aged, 80 and over ; Longitudinal Studies

Objective To explore whether hepatocyte Perilipin-2(Plin2)is involved in the development of fatty liver related to comparative gene identification-58(CGI-58)deficiency mice and compare the effects of Plin2 and Plin3 on lipid droplet formation and lipid accumulation.Methods Based on CGI-58Flox/Flox mice as animal model,the adeno-associated viruses targeting mouse liver,CGI-58 knockout and Plini2 knockdown were achieved by co-expression Cre protein and micro-RNA targeting Plin2(Mi-KD).Then CGI-58 deficiency mice were used as control(NC)to detect the differences in metabolic phenotype and liver pathology.AML-12 mouse hepatocytes were used as cellular model and interfered with siRNA to achieve Plin2/Plin3 knockdown in AML-12 cells.Lipid droplet formation and lipid accumulation were compared with Bodipy staining and enzyme colorimetry in basal condition or lipid-overloaded condition(OA inducement)after Plin2/Plin3 knockdown.Results Plin2 knockdown(Mi-KD)reduced PLIN2 protein level by＞99％in mouse livers.Mi-KD decreased hepatomegaly(P=0.019 5)and liver injury(P=0.000 4),while reduced the histological NAS score(P=0.000 2)and hepatic triglyceride content(P=0.016 6)in the CGI-58 deficiency female mice.Mi-KD prevented microvesicular hepatic steatosis in the CGI-58 deficient female mice.Plin3 knockdown significantly reduced the triglyceride content in basal condition of hepatocytes(P=0.001 4),and Plin2 knockdown just showed a decreased trend.Plin2 or Plin3 knockdown significantly reduced the triglyceride content separately in lipid-overloaded hepatocytes(P＜0.05).Conclusion Hepatocyte Plin2 is essential in the development of microvesicular hepatic steatosis caused by CGI-58 deficiency.Both Plin2 and Plin3 are involved in lipid droplet formation and lipid accumulation in hepatocytes,and Plin3 shows a stronger effect.

OBJECTIVE To investigate the improvement mechanism of caudatin on liver injury of rats. METHODS SD rats were randomly divided into blank group， model group， caudatin low-dose and high-dose groups （25， 50 mg/kg）， with 6 rats in each group. Diethylnitrosamine （DEN） was injected intraperitoneally three times per week for eight weeks to establish liver injury model of rats. At 5th week of modeling， the rats received relevant medicine or 0.5% sodium carboxymethylcellulose intragastrically for 4 weeks. The levels of liver function indexes [alanine transaminase （ALT）， aspartate transaminase （AST）， total protein （TP） and total bilirubin （TBI）] and inflammatory factors [interleukin （IL-6）， tumor necrosis factor α （TNF-α）， IL-1β] in serum were detected； the histopathological morphological changes of rat liver were observed； the positive protein expressions of nuclear factor κB （NF-κB） and 78 kDa glucose regulatory protein （Grp78） in liver tissue were also determined； the expressions of endoplasmic reticulum stress-related protein Grp78， C/EBP homologous protein （CHOP）， activating transcription factor 6 （ATF6） and inositol requiring enzyme 1α （IRE1α） and the level of protein kinase R-like endoplasmic reticulum kinase robertluoyi@126.com （PERK） in liver tissue were detected. RESULTS Compared with blank group， serum levels of ALT， AST， TBI， IL-6， TNF-α and IL-1β and positive expressions of NF-κB and Grp78 in liver tissue as well as protein expressions of Grp78， CHOP， ATF6 and IRE1α， PERK protein phosphorylation level were all increased significantly in model group （P＜0.05）， while the serum level of TP was decreased significantly （P＜0.05）. The disordered structure of liver lobule， swollen liver cells， unclear intercellular boundary were observed and accompanied by inflammatory cell infiltration. Compared with model group， most of the above indexes were significantly reversed in caudatin groups （P＜0.05）； the structure of hepatic lobule was relatively complete and clear， the cells were arranged orderly， and the infiltration of inflammatory cells was also reduced. CONCLUSIONS Caudatin has a significant improvement effect against DEN-induced liver injury in rats， the mechanism of which may be associated with inhibiting endoplasmic reticulum stress and inflammatory reaction.

Objective:To explore the team construction and treatment strategy of the Diabetic Foot-Multidisciplinary Team.Methods:The clinical data of 19 patients with severe ischemic diabetic foot treated by our Diabetic Foot-Multidisciplinary Team Center from Apr 2021 to Mar 2022 were collected, and the overall amputation rate, above-ankle major amputation rate, minor amputation rate and mortality, Diabetic Foot-Multidisciplinary Team consultation discipline participation rate and treatment participation degree were retrospectively analyzed.Results:Nineteen patients (15 males and 4 females) were enrolled, aged 26 to 94 (68.6±14.2). All were with severe ischemic diabetic foot ulcer:Rutherford grade 5 or up and dysfunction in 2 or more organs. Complications included arteriosclerosis obliterans of the lower extremities in 18 cases, heart diseases in 18, hypertension in 15, and renal insufficiencies in 10. The overall amputation rate was 36.8%, major amputation rate in 21.1%, minor amputation rate in 15.8%, and mortality rate was 15.8%. A total of 16 disciplines participated in Diabetic Foot-Multidisciplinary Team; the main participating disciplines were vascular surgery (19 times), endocrinology (12 times), and cardiology (11 times). The main treatment disciplines were vascular surgery (14 times), plastic surgery (3 times), and cardiology (2 times).Conclusion:For the diagnosis and treatment of diabetic foot, it is necessary to set up a multidisciplinary team as early as possible to control the causes of diabetic foot ulcer, prevent the recurrence of diabetic foot ulcer, reduce the mortality and amputation rate, and improve the quality of life of patients.

Objective:To analyze the medication law of Traditional Chinese Medicine (TCM) patent compounds for Alzheimer disease (AD) by using data mining method.Methods:The TCM compounds for the treatment of AD in the patent database were screened, and the frequency, clustering and association analysis were carried out with the help of TCM inheritance calculation platform, SPSS Statistics 21.0 and SPSS Modeler 18.0 software. The medication law was analyzed.Results:A total of 220 patent compounds were included, involving 361 kinds of Chinese materia medica; the top 10 high-frequency drugs were Acori Tatarinowii Rhizoma, Polygalae Radix, Ginseng Radix et Rhizoma, Chuanxiong Rhizoma, Astragali Radix, Lycii Fructus, Poria, Rehmanniae Radix PraeparataAngelicae Sinensis Radix, Salviae Miltiorrhizae Radix et Rhizoma; the most frequently used drugs were drugs for tonifying deficiency and promoting blood circulation to remove blood stasis; most of their properties belonged to warm, mild and cold; the tastes were mainly sweet, bitter and pungent; the meridians belonged to the five internal organs. 16 items of association data (4 combinations of two items and 12 combinations of three items) were obtained by association rule analysis, and the strongest correlation group was " Acori Tatarinowii Rhizoma- Polygalae Radix" and " Acori Tatarinowii Rhizoma- Chuanxiong Rhizoma- Polygalae Radix". Cluster analysis showed four prescription combinations and three pairs of drug compatibility, including the addition and subtraction structure of Kaixin Powder, Zuogui Pill, Bazhen decoction and so on. Conclusion:The core treatment principle of TCM patent compound treatment of AD is regulating and tonifying the five internal organs to treat its root, resolving phlegm and removing blood stasis to treat the symptoms, which accords with the theoretical basis of TCM in the treatment of AD, and can provide reference for clinical practice and new drug research and development.

Objective:To assess the efficacy and safety of 100 units of botulinum toxin A (BTX-A) intradetrusor injection in patients with overactive bladder.Methods:From April 2016 to December 2018, 17 tertiary hospitals were selected to participate in this prospective, multicenter, randomized, double-blind, placebo-controlled study. Two phases of study were conducted: the primary phase and the extended phase. This study enrolled patients aged 18 to 75 years who had been inadequately managed by anticholinergic therapy (insufficient efficacy or intolerable side effects) and had spontaneous voiding with overactive bladder. Exclusion criteria included patients with severe cardiac, renal and hepatic disorders, patients with previous botulinum toxin treatment for 6 months or allergic to BTX-A, patients with urinary tract infections, patients with urinary stones, urinary tract tumors, diabetes mellitus, and bleeding tendency. Eligible patients were randomly assigned to BTX-A group and placebo control group in a ratio of 2∶1. Two groups of patients received 20 intradetrusor injections of BTX-A 100U or placebo at the depth of the submucosal muscle layer respectively under cystoscope, including 5 injections at the base of the bladder, 3 injections to the bladder triangle, 5 injections each to the left and right walls and 2 injections to the top, sparing the bladder neck. As a placebo control group, patients received same volume of placebo containing no BTX-A and only adjuvant freeze-dried preparations for injection with the same method. A combination of gelatin, sucrose, and dextran served as adjuvants. Average micturition times per 24 hours, urinary incontinence (UI) episodes per day, average micturition volume per day, OAB symptom score(OABSS), and quality of life (QOL) score were recorded at baseline and the 2nd, 6th and 12th week after treatment. The primary efficacy endpoint was the change from baseline in the average micturition times per 24 hours at the 6th week after treatment. The secondary efficacy endpoints included the change from baseline in the average micturition times per 24 hours at 2nd and 12th week, as well as the change from baseline in the OABSS, QOL score, average frequency of urgency and UI episodes per day, urgency score, average micturition volume per day at 2nd, 6th and 12th week after treatment. Patients were followed for 12 weeks to assess adverse events (AEs). After assessed at week 12, if the micturition times has decreased less than 50% compared to baseline and the patient is willing to receive retreatment, then patients could enter the extended trial phase. In that phase, patients in both groups were injected with 100 units BTX-A from 12th week onwards and then followed up the same indicators for 12 weeks.Results:216 patients were enrolled in this trial (144 cases in the BTX-A group and 72 cases in the placebo control group). Baseline characteristics such as age (47.75±14.20 in the BTX-A group and 46.39±15.55 in the control group), sex (25 male/117 female in the BTX-A group and 10/61 in the control group), and disease duration (0.51 years in the BTX-A group and 0.60 years in the control group) were balanced between the two groups( P>0.05). A marked reduction from baseline in average micturition times per 24 hours was observed in all treatment groups at the 6th week and the reduction of the two groups was statistically different ( P<0.001 and P=0.008 respectively). Compared with the baseline, the average micturition times per 24 hours at the 6th week decreased from baseline by 2.40(0.70, 4.60)times for the BTX-A group and 0.70(-1.00, 3.30) times for the placebo control group respectively, and the difference between the two groups was considered to be statistically significant ( P=0.003). The change rates of average micturition times per 24 hours from baseline at the 6th week of the two groups were (16±22)% and (8±25)% respectively, and the difference between the two groups was statistically significant ( P=0.014). Compared with the baseline, the average micturition times per 24 hours at 2nd and 12th week decreased by 2.00(0.00, 4.00)and 3.30(0.60, 5.03)for the BTX-A group, 1.00(-1.00, 3.00)and 1.70(-1.45, 3.85)for the placebo control group respectively. The difference between two groups was considered to be statistically significant ( P=0.038 and P=0.012); the changes of average urgency times per day for the BTX-A group and the control group at the 2nd, 6th and 12th week were 2.00(0.00, 4.30)and 2.40(0.30, 5.00), 3.00(0.30, 5.70)and 0.70(-1.30, 2.70), 0.70(-1.30, 3.00) and 1.35(-1.15, 3.50), respectively. There were significant differences between two groups at the 2nd, 6th and 12th week, ( P=0.010, P=0.003 and P=0.025, respectively). The OABSS of the BTX-A group and the control group at the 6th week decreased by 1.00(0.00, 4.00)and 0.50(-1.00, 2.00) compared with the baseline, and the difference between the two groups was statistically significant ( P=0.003). 47 cases of BTX-A group and 34 cases of placebo control group entered the extended trial phase, and 40 and 28 cases completed the extended trial phase, respectively. The average micturition volume per 24 hours changed by -16.60(-41.60, -0.60)ml and -6.40(-22.40, 13.30)ml, (-35.67±54.41)ml and(-1.76±48.69)ml, (-36.14±41.51)ml and (-9.28±44.59)ml, (-35.85±43.35)ml and(-10.41±40.29)ml for two groups at the 12th, 14th, 18th and 24th week, and the difference between two groups was statistically significant at each follow-up time ( P=0.01, 0.006, 0.012 and 0.016, respectively). There was no significant difference in other parameters( P>0.05). However, adverse reactions after intradetrusor injection included increased residual urine volume (27 in the BTX-A group and 3 in the control group), dysuria (21 in the BTX-A group and 6 in the control group), urinary infection (19 in the BTX-A group and 6 in the control group), bladder neck obstruction (3 in the BTX-A group and 0 in the control group), hematuria (3 in the BTX-A group and 1 in the control group), elevated alanine aminotransferase (3 in the BTX-A group and 0 in the control group), etc. During the follow-up period, there was no significant difference in the other adverse events between two groups except the increase of residual urine volume( P<0.05). In the primary trial phase, among the 27 cases with increased residual urine volume in BTA group, only 1 case (3.70%) with PVR more than 300 ml; the PVR of 3 patients in the placebo group was less than 100 ml. The increase of residual urine volume caused by the injection could be improved or disappeared with the passage of time. Conclusions:Intradetrusor injection of Chinese BTX-A improved the average micturition times per 24 hours, the average daily urgent micturition times, OABSS, and average micturition volume per time, and reduced the adverse effects in patients with overactive bladder.Chinese BTX-A at dose of 100U demonstrated durable efficacy and safety in the management of overactive bladder.

Objective:To explore the expressions of miRNA-17-92 (miR-17-92) cluster and mitofusin 2 (MFN2) protein in endometrial cancer (EC) and their clinical significances.Methods:A total of 72 EC tissues, 36 endometrial lesions of patients with endometrial atypical hyperplasia, and 22 normal endometrial tissues from total hysterectomy for grade Ⅲ cervical intraepithelial neoplasia in the Second Affiliated Hospital of Shandong First Medical University from January 2008 to December 2014 were collected; at the same time, all patients' paraffin-embedded tissues were collected. Real-time quantitative polymerase chain reaction was used to detect the expression level of miR-17-92 in each tissue. Immunohistochemical SP method was used to detect the localization and expression level of MFN2 protein in each paraffin-embedded tissue. The correlation of miR-17-92 and MFN2 protein with clinicopathological features of EC patients was analyzed. Kaplan-Meier method was used to draw survival curve of patients with different miR-17-92 and MFN2 levels, and log-rank test was made; Cox proportional hazard regression model was used for multivariate survival analysis.Results:The relative expression of miR-17-92 in EC, atypical hyperplasia and normal endometrial tissues were 1.49±0.46, 1.01±0.30 and 0.69±0.20, respectively. The expression of miR-17-92 in EC tissues was higher than that in the other endometrial tissues, and the differences were statistically significant (both P < 0.01). The high-expression rates of MFN2 protein in EC, atypical hyperplasia and normal endometrial tissues were 20.8% (15/72), 39.4% (13/33) and 85.0% (17/20); the high-expression rate of MFN2 protein in EC tissue was lower than that in the other endometrial tissues, and the differences were statistically significant (both P < 0.012 5). In EC patients, the relative expression of miR-17-92 in patients with histological type Ⅱ was higher than that in patients with histological type Ⅰ ( P < 0.05); the relative expression of miR-17-92 in patients with myometrial invasion depth ≥1/2 were higher than that in patients with myometrial invasion depth <1/2 ( P < 0.05). The high-expression rate of MFN2 protein in patients with histological type Ⅰ was higher than that in patients with histological type Ⅱ ( P < 0.05); the high-expression rate of MFN2 protein in patients with International Federation of Gynecology and Obstetrics (FIGO) grade Ⅰ was higher than that in patients with FIGO grade Ⅱ and Ⅲ ( P < 0.05). When the EC patients were grouped according to the median relative expression of miR-17-92 (1.421), Kaplan-Meier survival analysis showed that the median overall survival (OS) time of the miR-17-92 low-expression group (36 cases) was not reached, and the high-expression group (36 cases) was 36 months (95% CI 32-42 months), and the difference in OS between the two groups was statistically significant ( P = 0.049); the median OS time of the MFN2 high-expression group (15 cases) was not reached, and the low-expression group (57 cases) was 38 months (95% CI 33-41 months), and the difference in OS between the two groups was statistically significant ( P = 0.046). Multivariate Cox regression analysis showed that the expression levels of miR-17-92 and MFN2 were independent influencing factors for the survival of EC patients ( HR = 3.10, 95% CI 1.36-7.07, P = 0.007; HR = 0.30, 95% CI 0.09-0.99, P = 0.048). Conclusion:The high-expression of miR-17-92 and low-expression of MFN2 protein in EC tissues may be involved in the occurrence and development of EC, and they can be used as indicators for judging the prognosis of EC patients.

OBJECTIVE：To investigate the effects and mechanism of Jidesh eng anti-venom tablet on local wound inflammation and systemic inflammatory response of snake bite patients. METHODS ：Totally 64 patients with snake bite admitted to our hospital during Jun. 2018-Jun. 2020 were randomly divided into control group and observation group ，with 32 cases in each group. Both groups received routine treatment ，such as debridement ，drainage，flushing，sealing，anti-venom，anti-infection，anti-fibrinolysis and anti-shock. Observation group was additionally given Jidesheng anti-venom tablet for internal and external use ，for consecutive 7 d. Related indexes of systemic inflammatory response ，local wound condition ，hospital stay ，laboratory indexes of important organs，coagulation function index ，wound inflammatory cell counts ，serum levels of inflammatory cytokines and chemokine ，the occurrence of ADR were compared between 2 groups. RESULTS ：After treatment ，most of related indexes of systemic inflammatory response （RR，HR and WBC ），local wound condition （local pain disappearance time ，wound detumescence time ）， hospital stay ，laboratory indexes of important organs （AST，ALT，Scr，BUN，CKB，CK-MB），coagulation function index （t-PA， PAI-1，TAT，SFMC），wound inflammatory cell （macrophages，neutrophils，lymphocytes）count，serum levels of inflammatory cytokines（TNF-α，IL-1，IL-6，hs-CRP，NF-κB）and chemokine （MCP-1，CXCL-8）in 2 groups were significantly better than before treatment （P＜0.05）；most indexes of observation group were significantly better than those of control group （P＜0.05）. No severe ADR was found in 2 groups. CONCLUSIONS ：Jidesheng anti-venom tablet as auxiliary treatment can significantly reduce the local wound inflammation and systemic inflammatory response of snake bite patients ；the mechanism is probably related to reducing the levels of chemokine MCP- 1 and CXCL- 8 and inflammatory cytokines hs-CRP and NF-κB.

Objective:To evaluate the diagnostic efficacies of computed tomography (CT), clinical manifestations and 2019 novel coronavirus (2019-nCoV) nucleic acid test results for the screening and diagnosis of novel coronavirus pneumonia.Methods:The clinical data of suspected cases with corona virus disease 2019 (COVID-19) visited fever clinic or stayed in emergency room of the First Affiliated Hospital of Nanchang University from January 23 to February 9, 2020 were collected. Totally 274 cases who met the inclusion criteria were enrolled. Four screening schemes including chest CT screening, scoring, chest CT and scoring series screening, chest CT and scoring parallel screening were designed. The statistical analysis was performed by using chi-square test. The sensitivities, specificities and the areas under the receiver operator characteristic curve of the four screening schemes were calculated, and the diagnostic efficacies were evaluated.Results:Among the 274 cases, 93.80%(257/274) presented with typical clinical symptoms, 47.81%(131/274) had epidemiological history, and the blood routine examination results of 45.26%(124/274) cases met the positive criteria of the scoring , and chest CT results of 43.07%(118/274) cases met the positive criteria of chest CT screening. The 2019-nCoV nucleic acid test positive rate of cases with epidemiological history was 30.53%(40/131), which was higher than that of cases without epidemiological history (9.09%(13/143)), and that of cases with typical imaging performance on chest CT was 40.68%(48/118), which was higher than cases without typical imaging performance (3.21%(5/156)). The differences between the above groups were both statistically significant ( χ2=20.150、60.468, respectively, both P<0.01). The positive rates of viral nucleic acid detection in cases with positive findings of chest CT screening, scoring, series screening, and parallel screening were 40.68%(48/118), 23.74%(47/198), 44.68%(42/94) and 23.87%(53/222), respectively, while those in cases with negative findings of the four screening schemes were 3.20%(5/156), 7.89%(6/76), 6.11%(11/180) and 0(0/52), respectively. The positive rates of the four screening schemes were all significantly different from that of viral nucleic acid detection ( κ=0.402, 0.100, 0.431, 0.106, all P<0.01). The chest CT screening method had a sensitivity of 90.57%and a specificity of 68.33%, and an area under the operating characteristic curve of the subject was 0.794, which had higher diagnostic value than those of the other three screening schemes. Conclusions:For the screening and diagnosis of COVID-19 cases, epidemiological history, positive 2019-nCoV nucleic acid test with ≥2 typical clinical manifestations have highly diagnostic value. On the basis of the preliminary screening of chest CT examination, flexible analysis of the diagnostic results could improve the diagnostic value of each detection method.