Objective: To explore the association between CDKAL1 rs35261542, FAIM2 rs 3205718, and VGLL4 rs 2574704 polymorphisms with childhood obesity and related metabolic phenotypes to provide evidence for personalized prevention and management strategies. Methods: Based on the 2023 Long term Nutritional Health Effects of Early Childhood Nutrition Package Intervention project, the study enrolled 1 078 children aged 5-7 years from four counties in Henan (Songxian and Ruyang countries) and Guizhou (Guiding and Fuquan countries) provinces. Using BMI Z scores, 87 overweight and obese(OVOB) children were selected and matched by sex, age, and BMI Z score with 117 normal weight controls. Participants were further stratified into four metabolic phenotype groups: metabolically healthy normal weight (MHNW, n =51), metabolically unhealthy normal weight (MUNW, n =66), metabolically healthy obesity (MHO, n =31) and metabolically unhealthy obesity (MUO, n =56) based on four conventional cardiometabolic risk factor (CR) criteria. Data were collected through questionnaires, anthropometric measurements, serum biochemical tests, and KASP genotyping. The distribution of three genetic polymorphisms ( CDKAL1 rs35261542, FAIM2 rs3205718, VGLL4 rs 2574704) across metabolic subgroups was analyzed. Multivariate Logistic regression models assessed associations between these polymorphisms and obesity/metabolic phenotypes. Results: Multivariate Logistic regression analysis showed that Homozygous mutant AA genotype of CDKAL1 rs 35261542 was positively associated with OVOB( OR =3.63), MHO ( OR =11.04), MUO ( OR = 4.88 ) ( P <0.05). Homozygous TT genotype of FAIM2 rs 3205718 increased OVOB risk ( OR =4.44, P <0.05) but showed no association with metabolic phenotypes ( P >0.05). Homozygous mutant TT of VGLL4 rs 2574704 reduced the risks of MHO and MUO ( OR = 0.30, 0.24， P <0.05). Cumulative genetic effects analysis demonstrated carriers of 1 or 2 risk genotypes of rs 35261542 and rs 3205718 had progressively higher OVOB risk ( OR =2.53, 20.79), and the combination of rs 35261542 and rs 2574704 increased risks for both MHO ( OR =8.50) and MUO ( OR =5.00) ( P <0.05). Conclusions The AA genotype of rs 35261542 ( CDKAL1 ) positively correlates with childhood obesity and metabolic abnormalities. The TT genotype of rs 3205718 ( FAIM 2) increases obesity risk but not metabolic phenotypes. The TT genotype of rs 2574704 ( VGLL 4) shows protective effects against metabolic dysfunction. Risk genotypes exhibit dosedependent cumulative effects on obesity and metabolic outcomes.

Objective To optimize the extraction process of Xuetong capsules and establish its quality control method. Methods The extraction process was optimized by orthogonal experiment using ethanol reflux method to investigate the effects of different factors on diphenylstilbene, aloin and extraction yield. The content of 5 anthraquinone compounds in Xuetong capsule was determined by HPLC. Results The optimal extraction process was to add 10 times ethanol, with an ethanol concentration of 70%, and extract 3 times, each time for 1 h; 5 components had a good linear relationship with peak area within a certain concentration range, r>0.999 7; The range of sample recovery rate was 93.66%-96.85%, RSD range of 1.48%-1.66%. The content determination results of the 5 components in three batches of Xuetong capsules were （0.632-0.641）, （0.660-0.681）, （1.968-1.991）, （2.547-2.580）, and （1.076-1.101） mg/g. Conclusion The method was accurate, reproducible, and highly feasible, which could be references for producing and improving the quality control standards of Xuetong capsules.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

Objective:To explore the effects of MiR-199a-3p on promoting the development of mycoplasma pneumonia via the silent information regulator 1(SIRT1)/nuclear factor-κB (NF-κB) pathway.Methods:Totally 80 SPF-grade BALB/c mice were randomly divided into a control group, a model group, a miR-199a-3p group, and a inhibitory group, with 20 rats in each group. Excep the control group, the model group was established as a mouse model of mycoplasma pneumoniae through a continuous nasal drip of a high-load mycoplasma pneumoniae bacterial solution for 3 days. After modeling, mice in each group had tail vein injections. The miR-199a-3p group and the inhibitory group mice were injected with agomir solution and antagonir solution, respectively. And the model group and control group mice were injected with the same amount of physiological saline through the tail vein. After the experiment, mice of all groups were killed and their blood was collected from the eyeballs, and interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) levels were detected by the enzyme-linked immunosorbent assay method. Subsequently, the lung tissues of the mice were taken for HE staining to observe pathological changes in the lung tissue. Real-time fluorescence quantitative PCR was used to detect miR-199a-3p gene expression levels in lung tissue, and Western Blot was used to detect SIRT1/NF-κB signaling pathway protein expression in lung tissue. Results:Compared with the model group, IL-4, IL-6, IL-1β, and TNF-α levels in the serum of the miR-199a-3p group were decreased, with a significant difference ( P < 0.05). The HE staining results showed that the lung tissue structure of the control group mice was nearly normal and there was no alveolar exudation or inflammatory response. The other three groups all had varying degrees of alveolar interstitial widening, alveolar exudation, and inflammatory cell infiltration. Compared with the model group, miR-199a-3p gene and SIRT1 protein expression levels in the miR-199a-3p group increased, with a significant difference (all P < 0.05). While NF-κB protein expression levels in the miR-199a-3p group decreased, there was a significant difference ( P < 0.05). Conclusions:The expression of miR-199a-3p gene is reduced in the lung tissue of mycoplasma pneumoniae mice. Increasing the level of miR-199a-3p gene has a protective effect on lung tissue damage in mycoplasma pneumoniae mice through anti-inflammatory effects, and its mechanism may be related to its regulatory effect on the SIRT1/NF-κB pathway.

Extracellular vesicles (EVs) are phospholipid bilayer vesicles actively secreted by cells, that contain a variety of functional nucleic acids, proteins, and lipids, and are important mediums of intercellular communication. Based on their natural properties, EVs can not only retain the pharmacological effects of their source cells but also serve as natural delivery carriers. Among them, plant-derived nanovesicles (PNVs) are characterized as natural disease therapeutics with many advantages such as simplicity, safety, eco-friendliness, low cost, and low toxicity due to their abundant resources, large yield, and low risk of immunogenicity in vivo. This review systematically introduces the biogenesis, isolation methods, physical characterization, and components of PNVs, and describes their administration and cellular uptake as therapeutic agents. We highlight the therapeutic potential of PNVs as therapeutic agents and drug delivery carriers, including anti-inflammatory, anticancer, wound healing, regeneration, and antiaging properties as well as their potential use in the treatment of liver disease and COVID-19. Finally, the toxicity and immunogenicity, the current clinical application, and the possible challenges in the future development of PNVs were analyzed. We expect the functions of PNVs to be further explored to promote clinical translation, thereby facilitating the development of a new framework for the treatment of human diseases.

Objective:To compare the effect of intracorporeal anastamosis and extracorporeal anastamosis on abdominal infection associated with laparoscopic right hemicolectomy.Methods:The clinical data of 210 patients with colon cancer who underwent laparoscopic right hemicolectomy in Dalian Third Peoples′s Hospital, Liaoning Province from January 2015 to December 2019 were analyzed retrospectively.Among them, 79 patients underwent intracorporeal anastamosis (intracorporeal anastamosis group) and 131 patients underwent extracorporeal anastamosis (extracorporeal anastamosis group). The perioperative indexes and postoperative abdominal infection were compared between the two groups.Results:In intracorporeal anastamosis group, the intraoperative bleeding was (45.2±4.2) mL, the operative time was (161.3±22.4) min, the number of lymph node dissection was (30.8±9.6), the postoperative exhaust time was (3.3±1.2) d, and the postoperative hospital stay was (7.6±0.5) d. In extracorporeal anastamosis group was (42.1±5.0) mL, (167.3±26.7) min, (32.9±8.6), (3.4±1.0) d and (7.5±0.6) d, respectively, there were no significant difference between the two groups (t value were 0.417, 0.207, 0.829, 0.338 and 0.293, respectively; P value were 0.699, 0.845, 0.231, 0.734 and 0.802, respectively). In intracorporeal anastamosis group, the incidence of abdominal infection (with anastomotic fistula)was 13.9%(11/79), the incidence of abdominal infection (without anastomotic fistula)was 10.1%(8/79), and in extracorporeal anastamosis group was 1.5%(2/131)and 0.8%(1/131), the differences were statistically significant (χ 2=12.805, 10.238; P=0.003, 0.008). In intracorporeal anastamosis group, the incidence of respiratory system infection was 1.3%(1/79), the incidence of urinary system infection was 2.5%(2/79), the incidence of surgical incision infection was 1.3%(1/79). In extracorporeal anastamosis group was 3.1%(4/131), 0.8%(1/131) and 3.1%(4/131), respectively.There were no significant difference between the two groups (χ 2 value were 0.662, 0.420 and 0.662, respectively; P value were 0.364, 0.587 and 0.364, respectively). Conclusion:Laparoscopic right hemicolectomy with intracorporeal anastamosis and extracorporeal anastamosis have the same surgical effect, but intracorporeal anastamosis may increase the risk of postoperative abdominal infection.

Objective:To compare the effect of midcaudal combined approach and the cephalic middle approach in laparoscopic complete mesocolic excision (CME) in the treatment of right colon cancer complicated with incomplete intestinal obstruction.Methods:From January 2014 to January 2019, 90 patients with right colon cancer complicated with incomplete intestinal obstruction admitted to the Third People′s Hospital of Dalian were retrospectively analyzed.All patients underwent laparoscopic right hemicolectomy, CME plus D3 lymph node dissection.According to the choice of different surgical approaches, 44 patients were treated with the midcaudal combined approach (observation group) and the other 46 patients were treated with cephalic middle approach (control group). The intraoperative, postoperative and complications of the two groups were compared statistically.Results:Compared with the control group, the bleeding volume in the observation group was significantly reduced ((105.3±22.6) ml vs.(309.6±28.0) ml, t=13.698), the operation time was significantly shortened ((165.2±17.9) min vs.(219.5±21.5) min, t=8.327), and the differences were statistically significant (all P<0.05). There were no significant differences in the number of lymph nodes dissected ((21.4±7.8)vs.(20.4±6.6), t=0.534), the proportion of lymph nodes dissected≥12(86.4%(38/44)vs.84.8%(39/46), χ 2=0.208), the complications after operation(6.8%(3/44)vs.10.9%(5/46), χ 2=0.318), the length of hospital stay after operation ((11.8±1.6) d vs.(12.5±2.3) d, t=0.986), the difference was statistically significant (all P>0.05). Conclusion:It is safe and feasible to use the middle caudal approach in laparoscopic CME for right colon cancer complicated with incomplete ileus. Compared with the cephalic middle approach, it can reduce thebleeding volume and shorten the operation time.

Objective: To investigate the clinical outcomes and second-look arthroscopic findings after high tibial osteotomy (HTO) combined with medial meniscus posterior root (MMPR) repair. Methods: Twenty-five patients who underwent HTO combined with MMPR repair were subjected to second-look arthroscopy and retrospectively analyzed. Biplane HTO combined with MMPR repair was performed on these patients. Arthroscopic transtibial pullout repair was employed to repair the MMPR. The relative degree of the medial meniscus extrusion (MME) were measured. Cartilage regeneration and the healing of MMPR were evaluated at the time of second-look arthroscopy. Clinical outcomes were assessed based on Hospital for Special Surgery (HSS) scores and Lysholm scores. Results: The MMPRs were completely healed in 12 cases (48%), partially healed in 9 cases (36%), healed with scarring in 3 cases (12%), and no healed in 1 case (4%). Follow-up duration was 13.04±1.06 months (12-16 months). There were no statistically significant differences in the Kellgren-Lawrence classifications of the cases before and after surgery (χ²=0.786, P=0.675). The relative position of the mechanical axis of the lower extremity through the tibial plateau was 19.88%±6.44% preoperatively and 58.68%±7.71% after operation with significant difference (t=-18.561, P<0.001). The Lysholm scores was increased significantly from 34.76±3.62 points to 82.08±4.35 points after operation (t=-52.717, P<0.001). The HSS scores was increased significantly from 33.52±6.48 points to 81.52±4.79 points after operation (t=-38.685, P<0.001). The degree of MME was changed from 51.12%±13.55% to 50.48%±15.15% without statistical difference (t=0.550, P=0.588) . The comparison between different degrees of healing groups revealed no statistical differences in all variables (P>0.05). The comparison between different degree of cartilage regeneration groups revealed no statistical differences in all variables (P>0.05). Conclusion HTO combined with MMPR repair can effectively improve the lower limb alignment and patients' symptoms with a satisfactory healing rate of MMPR. The effects of post-root repair after a short period is not obvious. The longer-term clinical effects is worthy of further observation.

Ischemia-reperfusion injury is one of the main causes of complications related to liver transplantation, hepatectomy, trauma and hemorrhagic shock. The cells of ischemia and hypoxic injury release of injury-related molecular patterns, lead to the activation of immune cells and cytokine, which further aggravates the inflammatory response and enlarges the injury. It’s indicated that injury-related molecular patterns, liver resident immune cells and cytokines play a key role in promoting inflammation and liver ischemia-reperfusion injury. However, recent studies suggested that the ischemia cells and cytokines played acomplex role in this process. Relevant progresses were reviewed in this article.