OBJECTIVES: To analyze the disease burden of melanoma among middle-aged and elderly populations in China, and to predict the future trend. METHODS: Data from the Global Burden of Disease (GBD) 2021 were utilized to collect incidence and mortality rates of melanoma, disability-adjusted life years (DALYs), and corresponding age crude rates among the middle-aged and elderly population in China during 1990 and 2021. Additionally, the estimated annual percentage change (EAPC) was employed to assess the temporal trends. Age-period-cohort (APC) and Bayesian age-period-cohort (BAPC) models were utilized to compute age, period, and cohort effects on incidence and mortality rates of melanoma, as well as to predict future trends up to 2035. RESULTS: During 1990-2021, the incidence rate of melanoma for males was higher than that for females among the middle-aged and elderly population in China, and the overall incidence rate increased annually with an EAPC of 2.13 (1.90-2.36), while the overall mortality rate and DALY rate showed a declining trend with an EAPC of －0.28 (－0.41-－0.15) and －0.54 (－0.68-－0.41), respectively. The results of the APC model analysis revealed that age effects on both incidence and mortality rates of melanoma in China's middle-aged and elderly population were significant, with both increasing with age. Period and cohort effects showed an upward trend for incidence rates but a downward trend for mortality rates. Moreover, the period and cohort effects for mortality rates were not significant among females. In the BAPC prediction model, the number of incidences of melanoma in middle-aged and elderly people in China would increase dramatically. By 2035, the number of incidence cases is expected to reach approximately 9600 (males) and 10 300 (females), corresponding to an incidence rate of 2.66/10⁵ and 2.67/10⁵, respectively. The number of deaths is projected to be about 2600 (males) and 3500 (females) by 2035, corresponding to a mortality rate of 0.72/10⁵ and 0.91/10⁵, respectively. CONCLUSIONS The disease burden of melanoma among the middle-aged and elderly population in China remains substantial and is expected to increase over the next decade.
Humans ; Melanoma/mortality* ; China/epidemiology* ; Aged ; Middle Aged ; Male ; Female ; Incidence ; Disability-Adjusted Life Years ; Bayes Theorem ; Cost of Illness ; Skin Neoplasms/epidemiology*

Diabetic foot ulcers (DFUs) is one of the severe complications of diabetes, which is difficult to treat and associated with a high risk of amputation. Current guidelines recommend that the plane of amputation be comprehensively assessed based on the degree of tissue necrosis and plane of vascular occlusion in DFUs, the age and systemic organ function of patients. However, the survival and quality of life of DFU patients suffering from amputation will be significantly affected. After reviewing the national and international literature and summarizing the experience of limb-salvage treatment of DFUs, we believe that free flap transplantation is of great significance for limb-salvage treatment to some patients with severe DFUs. In addition, this article also focus on analyzing the indications and perioperative precautions for patients with DFUs undergoing free flap transplantation.

Objective To explore the occurrence and influencing factors of delayed cerebral ischemia(DCI)after clipping surgery in patients with anterior circulation aneurysms.Methods A retrospective analysis was conducted on 248 patients with anterior circulation aneurysms who underwent clipping surgery in our hospital from May 2018 to May 2023.The patients were divided into the DCI group and the non-DCI group based on the occurrence of DCI after surgery.A1∶1 propensity score matching method was used to match 54 patients in the DCI group with 54 patients in the non-DCI group.The random forest algorithm was employed to rank the importance of factors influencing DCI.Multivariable Logistic regression analysis was performed to identify independent risk factors for DCI,and the E-value method was used to assess the sensitivity of model parameters.A DCI risk prediction model was constructed,and the discriminative ability and accuracy of the model were evaluated using ROC curve and Hosmer-Lemeshow test.Results After matching,no statistically significant differences were observed between the two groups in terms of gender,age,body mass index(BMI),smoking history,alcohol consumption,history of cerebral infarction,history of subarachnoid hemorrhage,aneurysm location,and maximum aneurysm diameter(all P＞0.05).Compared to the non-DCI group,the DCI group had a higher proportion of modified Fisher grade 3-4 and Hunt-Hess grade IV patients,higher levels of D-dimer(D-D),total bilirubin(TBIL),TNF-α,high-sensitivity C-reactive protein(hs-CRP),IL-6,and glutamate,and lower levels of immunoglobulin(Ig)G,IgM,and IgA(all P＜0.05).The random forest analysis showed that including Hunt-Hess grade,modified Fisher grade,D-D,glutamate,TB1L,IgA,and IL-6 in the multivariable regression analysis achieved the lowest out-of-bag error rate.Multivariable analysis revealed that Hunt-Hess grade Ⅳ,modified Fisher grade 3-4,D-D,glutamate,TBIL were independent risk factors for DCI,while IgA was a protective factor(all P＜0.05).The risk prediction equation was as follows:P=1/[1+e(3.142+1.307 Hunt-Hess grade+1.086 modified Fisher grade+0.976 D-D+0.748 glutamate+0.552 TBIL-0.524 IgA+0.487 IL-6)].A patient with a risk probability P＞0.65 was considered to have a postoperative occurrence of DCI.The ROC curve results showed that after internal validation,the AUC of the model was 0.869(95％CI:0.836-0.897),with sensitivity and specificity of 81.08％and 97.34％,respectively,indicating good discrimination of the model.The Hosmer-Lemeshow test results showed that after internal validation,the goodness of fit of the model wasx2=4.219,P=0.561,indicating good fit of the model.Conclusions Hunt-Hess grade Ⅳ,modified Fisher grade 3-4,D-D,glutamate,TBIL,and IL-6 are identified as independent risk factors for DCI,while IgA is a protective factor.The predictive model constructed using these factors demonstrates high accuracy and discriminative ability.A risk probability P＞0.65 indicates a postoperative occurrence of DCI in patients.

BACKGROUND: Vaccination has been shown effective in controlling the global coronavirus disease 2019 (COVID-19) pandemic and reducing severe cases. This study was to assess the flare and change in disease activity after COVID-19 vaccination in patients with stable rheumatoid arthritis (RA). METHODS: A prospective cohort of RA patients in remission or with low disease activity was divided into a vaccination group and a non-vaccination group based on their COVID-19 vaccination status. Each of them was examined every 3 to 6 months. In the vaccination group, disease activity was compared before and after vaccination. The rates of flare defined as disease activity scores based on 28-joint count (DAS28) >3.2 with ΔDAS28 ≥0.6 were compared between vaccination and non-vaccination groups. RESULTS: A total of 202 eligible RA patients were enrolled. Of these, 98 patients received no vaccine shot (non-vaccination group), and 104 patients received two doses of vaccine (vaccination group). The median time interval from pre-vaccination visit to the first immunization and from the second dose of vaccine to post-vaccination visit was 67 days and 83 days, respectively. The disease activity scores at pre-vaccination and post-vaccination visits in the vaccination group patients were similar. At enrollment, gender, RA disease course, seropositivity, and disease activity were comparable across the two groups. Flare was observed in five (4.8%) of the vaccination group patients and nine (9.2%) of the non-vaccination group patients at post-vaccination assessment ( P  = 0.221). In terms of safety, 29 (27.9%) patients experienced adverse events (AEs) after vaccination. No serious AEs occurred. CONCLUSIONS COVID-19 vaccinations had no significant effect on disease activity or risk of flare in RA patients in remission or with low disease activity. Patients with stable RA should be encouraged to receive the COVID-19 vaccination.
Humans ; Arthritis, Rheumatoid ; Cohort Studies ; COVID-19/prevention & control* ; COVID-19 Vaccines/adverse effects* ; East Asian People ; Prospective Studies ; Vaccination/adverse effects*

Objective:To explore the carrier status of carbapenems-resistant Klebsiella pneumoniae (CRKP) plasmids in burn patients and analyze the correlation of these plasmids with the transmission of CRKP. Methods:A retrospective observational study was conducted. A total of 26 CRKP strains, which were isolated from the clinic-related samples of 22 burn patients (with 20 males and 2 females, aged (42±16) years) admitted to the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from January to December 2017, were collected and individually numbered. The plasmids of the strains were extracted by alkali lysis. After determination of the plasmid concentration by a nucleic acid concentration detector, the agarose gel electrophoresis was used to visualize the bands, and rough plasmids typing was performed. The plasmid of the smallest numbered CRKP in each plasmid type was transformed into competent Escherichia coli ( E. coli) strain Top10 (hereinafter referred to as TOP10 strain). The growth of each transformed strains and a Top10 strain cultivated in ampicillin containing Luria-Bertani (LB) agar medium overnight was observed, and the proportion of successful transformation was calculated. The plasmids from the smallest numbered plasmid carrying CRKP strain of successfully transformed Top10 strains (hereinafter referred to as the smallest successfully transformed strain) and correspondingly numbered CRKP were extracted, and then, the agarose gel electrophoresis was used to visualize the bands. Aforementioned successfully transformed strains and a TOP10 strain were used for the antimicrobial susceptibility testing with 17 antibiotics commonly used in clinic. The plasmid from the smallest successfully transformed strain was sequenced using the next-generation sequencing technology. Bioinformatics analyses such as protein-coding gene prediction and protein sequence alignment were performed successively. The sequence was subsequently named pKP03-NDM1 according to the carrying of drug resistance gene. According to the whole genome sequence of the plasmid carried by the smallest successfully transformed strain, the polymerase chain reaction, agarose gel electrophoresis, and gene sequencing were used to detect the New Delhi metallo-beta lactamase-1 ( blaNDM-1) of plasmids in the remaining 25 strains of CRKP. The ST typing in multilocus sequence typing of 26 strains of CRKP was analyzed based on the literature. Results:Plasmids were successfully extracted from 26 CRKP, with mass concentrations ranging from 19.3 to 189.8 ng/μL. Each of the 26 CRKP carrying plasmids showed at least one band longer than 2 500 bp in the agarose gel electrophoresis, which were roughly divided into 6 patterns of A, B, C, D, E, and F. After overnight cultivation, no growth of strains was observed in LB agar medium containing ampicillin inoculated with the TOP10 strain or TOP10 strains transformed by the plasmid of CRKP patterning A, B, D, or E. In contrast, TOP10 strains transformed by the pattern C plasmid from NO.3 CRKP and the pattern F plasmid from NO.15 CRKP resulted in numerous colony growths, and those transformed strains were named as TOP10-pKP03 and TOP10-pKP15, respectively. The proportion of successful transformation was 1/3. The plasmid carried by TOP10-pKP03 showed a single band in the agarose gel electrophoresis, which was the same size as the largest band of the plasmid from NO.3 CRKP. The TOP10 strain was sensitive to the 17 antibiotics commonly used in clinic. TOP10-pKP03 and TOP10-pKP15 were resistant to penicillins, cephalosporins, and carbapenems but remained sensitive to monocyclic β-lactam, aminoglycosides, quinolones and tigecycline. The full length of the plasmid carried by TOP10-pKP03 was 41 190 bp. In addition to blaNDM-1, this plasmid carried bleMBL, T4SS, bleomycin resistance gene, conjugation transfer elements, and relaxase, etc. The plasmid showed 99% nucleotide identity similarity and the same length to the plasmid pJN24NDM1 extracted from an E. coli isolate JN24. Totally 16 (61.5%) CRKP were confirmed to carrying blaNDM-1 gene, among the ST typing of the 16 strains, 11 strains were ST11, while ST215, ST260, ST395, ST2230, and new ST had 1 strain each. Among the ST typing of 10 blaNDM-1-negative CRKP, 8 strains were ST11, while ST395 and ST2230 had 1 strain each. Conclusions:A blaNDM-1 gene carrying plasmid pKP03-NDM1 was extracted and sequenced from CRKP isolated from burn patients, with a high plasmid carrying rate. Meanwhile, this plasmid may mediate inter-CRKP and CRKP- E. coli horizontal transfer of blaNDM-1, leading to transmission of antimicrobial resistance.

Objective:To explore the clinical application value of MRI in quantitative evaluation of anterior cruciate ligament mucoid degeneration (ACL-MD).Methods:From March to July 2020, 40 patients who were scheduled to undergo arthroscopic treatment were prospectively collected in the First Affiliated Hospital of Kunming Medical University.The anterior cruciate ligament tissue from the lateral edge of the tibial end was taken during the operation. Based on the pathologicalre sults, the patients were divided into the ACL-MD group ( n=19) and the normal group ( n=21). The sagittal plane three-dimensional steady-state rapid precession (3D-FIESTA), T 1 mapping, T 2 mapping, and T 2* mapping were performed before the knee joint surgery, and the scanned images were post-processed and analyzed to measure the T 1, T 2, and T 2* values of the tibial end of the anterior cruciate ligament.The relaxation time of the ACL-MD group and the normal group was compared using independent sample t test. The ROC curve was drawn using each parameter and the areas under the curve (AUC) for the diagnosis of ACL-MD were obtained.DeLong test was used to compare the differences of AUCs. Results:The T 1 [(1 291.9±273.4) ms], T 2 [(54.8±10.6) ms], and T 2* values [(30.6±6.4) ms] of anterior cruciate ligaments in the ACL-MD group were significantly higher than those in the normal group [ (1 087.0±121.0), (44.8±7.1), (20.4±4.8) ms; t=3.011, 3.473, 5.658, all P<0.001]. The AUCs of T 1, T 2, T 2* were 0.747, 0.764, 0.912, sensitivity of 63.2%, 63.2%, 100%, and the specificity of 100%, 95.2%, 76.2% in diagnosing ACL-MD. The AUC of the T 2* value was higher than those of the T 1 and T 2 values, and the differences were statistically significant ( Z=1.734, 2.162, P=0.043, 0.031). Conclusion:T 1, T 2, T 2*values measured by MRI quantitative imaging have high performance in assessing knee joint ACL-MD, and T 2* value has the largest AUC and the highest diagnostic efficiency.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) protein system, as an emerging gene editing system, can be divided into class 1 and class 2 systems according to the number of Cas protein. The CRISPR/Cas9 in class 2 system can cleave target nucleic acid only with the help of Cas9 protein and single-stranded guide RNA, which is currently the most widely used CRISPR/Cas system. In addition to gene editing in the treatment of genetic diseases, a variety of CRISPR/Cas system derived technologies have vast application prospect in the fields of disease-related gene screening, gene expression regulation, and rapid detection, prevention, and control of pathogens. This article summarizes the discovery process of CRISPR/Cas system and applications of several major CRISPR/Cas derived technologies, aiming to provide a reference for researchers in the field of life science.

Objective:To explore the influence of human amniotic mesenchymal stem cells (hAMSCs) on the in vivo and in vitro regulation of macrophage phenotypes and inflammatory factors associated with wound healing of full-thickness skin wounds in mice.Methods:Fresh amniotic membrane discarded from full-term delivery by 5 healthy pregnant women in the Department of Obstetrics and Gynecology of the Affiliated Hospital of Zunyi Medical University was used for the isolation and culture of hAMSCs by enzyme digestion method. The third passage of cells was used for identification of adipogenic and osteogenic differentiation. The fourth passage of cells was used for identification of hAMSCs surface markers. Ten C57BL/6 mice (all male, aged 6 to 8 weeks, the same gender and age below) were selected for extracting mouse peritoneal macrophages by intraperitoneal lavage, and M1-type macrophages were induced by Dulbecco′s modified eagle medium (DMEM) medium containing interferon-γ. The M1-type macrophages were divided into hAMSCs+ macrophage group and macrophage alone group. Then 1×10 4 hAMSCs/per well of fourth passage were added to macrophage in hAMSCs+ macrophage group and cultured in 2 mL DMEM medium for routine culture. In macrophage alone group, each well was only added with 2 mL DMEM medium for routine culture. On day 1 and 7 in culture, the content of interleukin-12 (IL-12), arginase 1, and IL-10 in the cell culture supernatant of the 2 groups were detected by enzyme-linked immunosorbent assay with sample number of 6/per group. (2) Full-thickness skin wound model was reproduced in the back of 56 C57BL/6 mice, which were divided into hAMSCs group and phosphate buffer solution (PBS) group using the random number table, with 28 mice in each group. Mice in hAMSCs group were subcutaneously injected with 100 μL of cell suspension containing 1×10 7 hAMSCs per mL in PBS suspension along the wound edge. While mice in PBS group were only subcutaneously injected with 100 μL PBS along the wound edge. On post injection day (PID) 1, 3, 7, and 14, 7 mice in the two groups were sacrificed respectively. Histopathological observation was performed with hematoxylin-eosin staining. The expressions of macrophage surface markers [CD68 and inducible nitric oxide synthase (iNOS) double positive cells and CD68 and arginase 1 double positive] in the wounds were detected by immunofluorescent staining. The mRNA expressions of IL-10, macrophage inflammatory protein 1α (MIP-1α), and MIP-2 in the wounds were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with analysis of variance for factorial design, t test, and Bonferroni correction. Results:(1) On day 1 in culture, the content of IL-12 and arginase 1 in the cell culture supernatant of the two groups were similar ( t=0.448, 0.536, P>0.05), and the content of IL-10 in the cell culture supernatant of hAMSCs+ macrophage group was significantly lower than that in macrophage alone group ( t=14.722, P<0.01). On day 7 in culture, the content of IL-12 in the cell culture supernatant of hAMSCs+ macrophage group was significantly lower than that in macrophage alone group ( t=13.226, P<0.01), and the content of arginase 1 and IL-10 was significantly higher than that in macrophage alone group ( t=30.172, 31.406, P<0.01). (2) On PID 1, a large number of inflammatory cells infiltration were observed in the skin wounds of both groups. On PID 3, the inflammatory cells infiltration in the skin wounds increased in both groups, and the inflammatory cells infiltration in hAMSCs group was less than that in the PBS group. On PID 7, the inflammatory cells infiltration in the wounds decreased in both groups, and the inflammatory cells infiltration in hAMSCs group was less than that in the PBS group. On PID 14, no obvious inflammatory cells infiltration was observed in the wounds in the two groups. (3) On PID 1 and 14, the percentages of CD68 and iNOS double positive cells and CD68 and arginase 1 double positive cells in the wounds were similar in the two groups ( t1 d=0.134, 0.693, t14 d=1.146, 2.585, P>0.05). On PID 3 and 7, the percentages of CD68 and iNOS double positive cells in the wounds in hAMSCs group were significantly lower than those of PBS group ( t=6.396, 4.787, P<0.01), while the percentages of CD68 and arginase 1 double positive cells were significantly higher than those of PBS group ( t=3.928, 4.473, P<0.01). (4) On PID 1, the mRNA expressions of IL-10 in the wounds of mice in the two groups were similar ( t=2.005, P>0.05). On PID 3, 7, and 14, the mRNA expressions of IL-10 in the wounds of mice in hAMSCs group were significantly higher than those of PBS group ( t=7.758, 124.355, 80.823, P<0.01). On PID 1, 3, 7, and 14, the mRNA expressions of MIP-1α and MIP-2 in the wounds of mice in hAMSCs group (0.341±0.212, 0.648±0.004, 0.611±0.106, 0.763±0.049, 1.377±0.099, 1.841±0.042, 1.181±0.035, 0.553±0.028) were significantly lower than those of PBS group (3.853±0.035, 6.914±0.163, 3.648±0.113, 2.250±0.046, 11.119±0.495, 8.634±0.092, 5.722±0.021, 4.862±0.036, t=43.198, 101.904, 51.845, 58.231, 51.074, 177.501, 291.752, 251.614, P<0.01). Conclusions:hAMSCs demonstrates biological effects of promoting the transformation of M1-type macrophages into M2-type macrophages in full-thickness skin wounds of mice. They can up-regulate the expression of anti-inflammatory and anti-fibrotic factor IL-10, and down-regulate the expression of important inflammation mediated factors MIP-1α and MIP-2.

Objective:To investigate the clinical effects of modified fascia flap from cutaneous branch of dorsal metacarpal artery in repairing the wound at the proximal and middle finger segments.Methods:From January 2017 to September 2018, 12 patients with wounds at the proximal and middle finger segments were admitted to the Affiliated Hospital of Zunyi Medical University, including 8 males and 4 females, aged 35-70 years. The areas of wounds ranged from 3.4 cm×2.4 cm to 6.5 cm×4.0 cm. The modified fascia flaps from cutaneous branch of dorsal metacarpal artery were resected to repair the wounds, with the size ranging from 3.5 cm×2.5 cm to 6.7 cm×4.1 cm. The flap donor sites of 5 patients were repaired with direct intermittent suture, the flap donor sites of 4 patients were repaired with full-thickness skin grafts from ipsilateral medial forearm, and the flap donor sites of 3 patients were repaired with wrist pedicled flaps. The survival of the flaps was recorded. Healing of donor site and recipient site was followed. The hand functions were evaluated with trial standard for the evaluation of the functions of the upper limbs of the Hand Surgery Society of the Chinese Medical Association.Results:All the flaps survived in 12 cases. During 3 to 12 months of follow-up, the flaps recovered satisfactorily in texture and shape. The donor sites of 11 patients were healed, and the skin graft edge area was partially necrotic in the other patient but healed later after dressing change. The distances of two-point discrimination of the patients ranged from 5.6 to 9.0 mm. Hand functions were evaluated as excellent in 5 cases, good in 4 cases, and fair in 3 cases.Conclusions:Modified fascia flap from cutaneous branch of dorsal metacarpal artery for repairing the wounds at the proximal and middle finger segments has reliable blood supply. The operation is simple and safe with short course of treatment, which is worthy of clinical promotion.

With the change of disease spectrum and the progress in the aging of society, chronic wound has gradually become one of the major diseases that threaten human health, and also one of the major economic burdens of family and society. According to the different etiology, the pathogenesis of chronic wound is different, including both systemic factors and local factors. The treatment of chronic wound is a multi-disciplinary integrated treatment process, including internal medicine treatment, surgical treatment, vascular interventional therapy, platelet-rich plasma treatment, and biological therapy, etc. Each treatment regimen has its own indications and pros and cons. To make a treatment regimen, a combination of a variety of options should be chosen according to the patient′s wound conditions. The traditional chronic wound treatment mode is multi-disciplinary team (MDT) treatment mode, which requires the participation of surgeons from multiple departments such as intervention department, plastic surgery department, orthopedics department, etc., and it is also the mainstream mode for treating chronic wound in western countries. According to the domestic medical situation and the experience of our department, we put forward the integrated surgical wound treatment (ISWT) mode, that is to integrate multiple surgical techniques of wound treatment. Compared with the traditional MDT treatment mode, to apply the ISWT mode can make a more reasonable treatment plan, improve the efficiency of diagnosis and treatment, shorten the hospitalization period, and improve the diagnosis and treatment ability of the team. With the increasing incidence of chronic wound, the ISWT mode needs to be further explored and improved, and the team needs more specialized experts to join in.