Objective: To establish and identify hepatocyte-specific transmembrane protein 121 ( Tmem121 ) knockout mice． Methods: The hepatocyte-specific Tmem121 knockout mice ( Tmem121flox / flox / Cre，Tmem121ΔHep) were obtained by crossbreeding of Tmem121flox / + / Cre and Tmem121flox / flox mice，which were generated using the CRISPR / Cas9 and Cre / Loxp systems．The genotype was verified by PCR using genomic DNA extracted from mouse tails as template．The growth，reproduction and organ development of both control and knockout mice were ob- served and analyzed．PCR and Western blot methods were performed to assess the knockout efficiency of Tmem121 in mouse primary hepatocytes．CellMaskTM Deep Red plasma membrane staining was employed to compare the mor- phological differences in primary hepatocytes between control and knockout mice． Results: Tmem121flox / flox / Cre mice were successfully obtained according to genotype identification analysis，and there were no significant differ- ences between control and knockout mice in body mass，reproductive ability，growth and development of liver．The specific knockout of Tmem121 gene in primary hepatocytes did not significantly affect the morphological structure or pathological characteristics of liver tissue．However，compared to the control group，the levels of Tmem121 mRNA and protein in the primary hepatocytes of the knockout group were significantly reduced ( P ＜0. 01) ．CellMaskTM Deep Red plasma membrane staining indicated that the proportion of binucleated hepatocytes in Tmem121-deficient mice significantly increased ( P＜0. 05) ，while the cell area was significantly reduced ( P＜0. 001) ． Conclusion Hepatocyte-specific Tmem121 knockout mice are successfully constructed，which provides an animal model for further exploration of the function and mechanism of Tmem121 gene in liver diseases．

This study was aimed at comparing the effects of storage conditions on the Brucella survival number in brain heart infu-sion medium,to provide experimental data support for the selection of storage conditions for Brucella strains.We created bacterial sus-pensions of seven strains of Brucella melitensis,Brucella abortus,and Brucella suis(denoted A,B,C,D,E,F,G)by using 30%glycerol brain heart infusion medium,then conducted stability testing.For accelerated stability tests,the bacterial solution was placed in a 37℃chamber for 1,2,3,or 4 weeks continuously,and the Brucella survival number at each time point was measured.For freeze-thaw stability testing,the bacterial solutions were frozen at-80℃,transferred to room temperature for 4 hours,and melted in one freeze-thaw cycle,and the Brucella survival number was measured for 2,4,6,and 8 freeze-thaw times.For long-term stability testing,the bacterial solutions were stored at-80℃,and the Brucella survival number was measured at various time points during the first,third,sixth,and twelfth months of storage.A line graph was plotted to demonstrate the changes in Brucella survival number at each time point in the three stability tests.IBM SPSS Statistics 22 software was used to assess statistically significant differences in Brucella survival number at each time point and the initial Brucella survival number.Accelerated stability tests showed that the Bru-cella survival number of the seven strains significantly decreased,and a statistically significant difference in Brucella survival number was observed after 1 week.In freeze-thaw stability tests,with increasing freeze-thaw times,the Brucella survival number of the seven strains showed a decreasing trend.Strains C and F showed statistically significant differences with respect to the initial Brucella sur-vival number after two times of freeze-thaw;strains A and D showed statistically significant differences with respect to the initial Bru-cella survival number after four times of freeze-thaw;strain G showed statistically significant differences with respect to the initial Bru-cella survival number after six times of freeze-thaw;and strain E showed statistically significant differences with respect to the initial Brucella survival number after eight times of freeze-thaw.The data for strain B showed abnormalities.In long-term stability testing,the Brucella survival number for strains C and F after storage for 1 month showed a statistically significant difference with respect to the initial bacterial solution.No statistically significant difference in Brucella survival number was observed for strains B and E after stor-age for 1,3,6,or 12 months.The data for strains A,D,and G showed abnormalities.The sterile 30%glycerol brain heart infusion medium maintained the Brucella survival number in an-80℃environment,and freezing and thawing multiple times should be avoided.Compared with the traditional freeze-drying method,this method is not only easy to perform but also avoids potential bio-safety hazards,and provides a reliable Brucella culture preservation scheme for researchers in related fields.

This study was aimed at comparing the effects of storage conditions on the Brucella survival number in brain heart infu-sion medium,to provide experimental data support for the selection of storage conditions for Brucella strains.We created bacterial sus-pensions of seven strains of Brucella melitensis,Brucella abortus,and Brucella suis(denoted A,B,C,D,E,F,G)by using 30%glycerol brain heart infusion medium,then conducted stability testing.For accelerated stability tests,the bacterial solution was placed in a 37℃chamber for 1,2,3,or 4 weeks continuously,and the Brucella survival number at each time point was measured.For freeze-thaw stability testing,the bacterial solutions were frozen at-80℃,transferred to room temperature for 4 hours,and melted in one freeze-thaw cycle,and the Brucella survival number was measured for 2,4,6,and 8 freeze-thaw times.For long-term stability testing,the bacterial solutions were stored at-80℃,and the Brucella survival number was measured at various time points during the first,third,sixth,and twelfth months of storage.A line graph was plotted to demonstrate the changes in Brucella survival number at each time point in the three stability tests.IBM SPSS Statistics 22 software was used to assess statistically significant differences in Brucella survival number at each time point and the initial Brucella survival number.Accelerated stability tests showed that the Bru-cella survival number of the seven strains significantly decreased,and a statistically significant difference in Brucella survival number was observed after 1 week.In freeze-thaw stability tests,with increasing freeze-thaw times,the Brucella survival number of the seven strains showed a decreasing trend.Strains C and F showed statistically significant differences with respect to the initial Brucella sur-vival number after two times of freeze-thaw;strains A and D showed statistically significant differences with respect to the initial Bru-cella survival number after four times of freeze-thaw;strain G showed statistically significant differences with respect to the initial Bru-cella survival number after six times of freeze-thaw;and strain E showed statistically significant differences with respect to the initial Brucella survival number after eight times of freeze-thaw.The data for strain B showed abnormalities.In long-term stability testing,the Brucella survival number for strains C and F after storage for 1 month showed a statistically significant difference with respect to the initial bacterial solution.No statistically significant difference in Brucella survival number was observed for strains B and E after stor-age for 1,3,6,or 12 months.The data for strains A,D,and G showed abnormalities.The sterile 30%glycerol brain heart infusion medium maintained the Brucella survival number in an-80℃environment,and freezing and thawing multiple times should be avoided.Compared with the traditional freeze-drying method,this method is not only easy to perform but also avoids potential bio-safety hazards,and provides a reliable Brucella culture preservation scheme for researchers in related fields.

Objective:To explore the trajectory of social vulnerability in first-episode stroke patients and analyze its influencing factors.Methods:Using convenience sampling, 210 stroke patients admitted to the Department of Cerebrovascular Diseases in The 3 rd Affiliated Hospital of CCUCM from January to December 2023 were selected as study subjects. The General Information Questionnaire, the Chinese version of the Social Vulnerability Index (SVI), Nutritional Risk Screening 2002, and the Social Support Rating Scale were used to conduct surveys at admission (T 1), 2 weeks post-onset (T 2), 1 month post-onset (T 3), 3 months post-onset (T 4), and 6 months post-onset (T 5). Latent Class Growth Model (LCGM) and univariate analysis were used for data processing. Results:A total of 176 valid consecutive questionnaires were collected, with an effective response rate of 83.81% (176/210). The SVI scores at T 1 to T 5 were (18.64±5.82), (19.97±6.42), (16.19±5.34), (15.98±5.61), and (16.12±4.42), respectively. Three latent classes of social vulnerability trajectories were identified among first-episode stroke patients, with average probabilities of 0.942, 0.956, and 0.932 for patients belonging to each latent class. The three classes were the high-level worsening group (30.1%, 53/176), the moderate-level improving group (52.3%, 92/176), and the low-to-moderate-level stable group (17.6%, 31/176). Age, living arrangement, self-rated personality type, activities of daily living after illness, presence of malnutrition, and social support were influencing factors for grouping the trajectories of social vulnerability in first-episode stroke patients ( P<0.05) . Conclusions:First-episode stroke patients exhibit three distinct trajectory types of social vulnerability from admission to 6 months post-onset, with variations in their social vulnerability trajectories. Age, living arrangements, self-rated personality types, activities of daily living after illness, presence of malnutrition, and social support are influencing factors for grouping social vulnerability trajectories in first-episode stroke patients. Clinical staff should closely monitor stroke patients in the high-level worsening group, promptly identify high-risk patients with social frailty, in order to reduce the impact of social vulnerability and provide proactive and targeted protective care.

ObjectiveTo investigate the effects and mechanism of Zuogui Jiangtang Tongmai prescription （ZJTP） on human umbilical vein endothelial cells （HUVECs） damaged by high glucose combined with lipopolysaccharide （LPS）. MethodThe survival rate of cells was determined by cell counting kit-8 （CCK-8）， and the level of tumor necrosis factor-α （TNF-α） was determined by enzyme-linked immunosorbent assay （ELISA） to determine the optimal injury concentration and action time of LPS， as well as the optimal action concentration of ZJTP drug-containing serum. HUVECs were divided into a blank control group， a model group， a ZJTP drug-containing serum group， and an SCFA mixed liquid group. ELISA was used to detect the level of endothelin-1 （ET-1）， nitric oxide （NO）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and TNF-α. Western blot was performed to detect the protein expression of G protein-coupled receptor43 （GPR43）， β-suppressor protein-2 （β-arrestin-2）， nuclear factor-κB suppressor α （IκBα）， and nuclear factor κB p65 （NF-κB p65）. The nucleation of NF-κB p65 was observed by immunofluorescence staining （IF）. The role of GPR43 in the regulation of inflammatory injury was observed by means of small interfering ribonucleic acid （siRNA）. The cells after intervention were divided into an empty carrier group， a ZJTP drug-containing serum group， a Si-GPR43 group， and a Si-GPR43 + ZJTP drug-containing serum group. The content of IL-1β， IL-6， and TNF-α was detected by ELISA. The protein expression of pathways was detected by Western blot. IF was used to observe the nucleation of NF-κB p65. ResultThe optimal molding condition was 1 mg·L^-1 LPS for 24 h. The optimal drug intervention condition was 5% ZJTP drug-containing serum for 24 h. Compared with the blank control group， the content of ET-1 in the model group was significantly increased， and the content of NO was significantly decreased （P<0.01）. The levels of inflammatory factors were significantly increased （P<0.01）. The expressions of GPR43 and IκBα were significantly decreased， while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly increased （P<0.01）. NF-κB p65 protein was transferred from the extranuclear to the intranuclear （P<0.01）. Compared with the model group， the content of ET-1 in the ZJTP drug-containing serum group was decreased， and the content of NO was increased （P<0.05）. The levels of inflammatory factors decreased （P<0.05）. The protein expressions of GPR43 and IκBα were increased， while the expressions of β-arrestin-2 and NF-κB p65 were decreased （P<0.05）. The amount of NF-κB p65 transferred from the intranuclear to the extranuclear decreased （P<0.01）. The mechanism study showed that compared with the Si-GPR43 group， the content of IL-1β， IL-6， and TNF-α were significantly decreased after treatment with ZJTP drug-containing serum （P<0.01）. The protein expressions of GPR43 and IκBα were significantly increased （P<0.01）， while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly decreased （P<0.01）. The amount of NF-κB p65 transferred from the extranuclear to the intranuclear decreased （P<0.01）. ConclusionZJTP has a protective effect on HUVECs with high glucose and LPS-induced inflammatory injury， which may be related to the regulation of GPR43/β-arrestin-2/IκBα/NF-κB pathway.

The Rainbow Model of Integrated Care(RMIC)is a new conceptual framework that integrates primary care principles,integrated care characteristics,and a triple aim framework based on the Rainbow Model,which helps researchers better understand the concept of integrated care from a primary care perspective and thus scientifically conduct integrated care practice programs.This paper reviews the emergence and development of RMIC,its conceptual framework,and its application in integrated care,with the aim of providing a guiding basis for improving the quality of integrated care and positively transforming the health care delivery model in China.

Objective Glucagon-like peptide-1 (GLP-1) has been reported to be effective in the treatment of nonalcoholic fatty liver disease (NAFLD). However, the molecular mechanism of GLP-1 on NAFLD is remained unclear. The present study was to detect whether the effect of GLP-1 on triglyceride (TG) content in hepatocytes is dependent on Foxos. Methods HepG2 cells were treated with palmitic/oleic acid for 24 h. The knockdown of Foxo1, Foxo3 was conducted through small interfering RNA (siRNA). Real time PCT (RT-PCR) was used to detect the changes of the SREBP1c and Acox2 genes in HepG2 cells after Foxo1/3 knockdown. Results As expected, palmitic/oleic acid increased TG concentration in HepG2 cells [(12.65 ± 1.32) μg/mg vs. (4.32 ± 0.54) μg/mg, P<0.05]. Addition of GLP-1 dose (10, 50, 100nmol/L) dependently lowered the TG content and reached plateau at 100 nmol/L of GLP-1 [TG(8.38±1.47) μg/mg]. The GLP-1 effect on TG remained after knocking down either Foxo1 [(9.09±1.34)μg/mg] or Foxo3 [(8.90± 1.60) μg/mg] alone, but not when knocking down Foxo1 and Foxo3 (Foxo1/3) together [(14.66±1.77)μg/mg]. Moreover, knocking down Foxo1/3 also abolished GLP-1 effect on SREBP1c and Acox2 expression. Conclusion GLP-1 can inhibit the synthesis of TG in hepatocytes depending on Foxo1 and Foxo3. Further studies are needed to explore the specific mechanisms.

BACKGROUNDSteroids inhibit osteogenic differentiation and decrease bone formation while concomitantly inducing adipose deposition in osteocytes. This leads to the fatty degeneration and necrosis of bone cells commonly seen in osteonecrosis of the femoral head. The peroxisome proliferator-activated receptor-γ (PPARγ) is an adipogenic transcription factor linked to the development of this disease and responsible for inducing adipogenesis over osteogenesis in bone marrow mesenchymal stem cells (BMSCs). The aim of this study was to assess whether adipogenic differentiation could be suppressed, and thus osteogenic potential retained, by inhibiting PPARγ expression in BMSCs.

METHODSCells from the bone marrow of New Zealand rabbits were treated with 10(-7) mol/L dexamethasone and infected with one of three small interference RNA (siRNA) adenovirus vectors (S1, S2, and S3) or non-targeting control siRNA (Con) and compared with dexamethasone-treated (model) and untreated (normal) cells. Cells were grown for 21 days and stained with Sudan III for adipocyte formation. At various time points, cells were also assessed for changes in PPARγ, osteocalcin (OC), Runx2, alkaline phosphatase (ALP) activity, and triglyceride (TG) content.

RESULTSDexamethasone-treated model and control groups showed a significant increase in fatty acid-positive staining, which was inhibited in cells treated with PPARγ siRNA-treated, similar to normal untreated cells. All three siRNA groups significantly inhibited PPARγ mRNA and protein, adipocyte number, and TG content compared with the dexamethasone-treated model and control groups, matching that seen in normal cells. OC and Runx2 mRNA and protein, as well as ALP activity, were significantly higher in cells treated with siRNA against PPARγ, similar to that seen in the normal cells. These osteogenic markers were significantly lower in the dexamethasone-treated cell cultures.

CONCLUSIONSThe siRNA adenovirus vector targeting PPARγ can efficiently inhibit steroid-induced adipogenic differentiation in rabbit BMSCs and retain their osteogenic differentiation potential.

Adenoviridae ; genetics ; Adipogenesis ; drug effects ; genetics ; Animals ; Cell Differentiation ; drug effects ; genetics ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; PPAR gamma ; genetics ; metabolism ; pharmacology ; RNA, Small Interfering ; Rabbits ; Steroids

Objective To translate LARS Score into Chinese and examine the reliability and validity of the LARS Score to predict patient bowel function.Methods A convenience sample of 40 Chinese rectal patients was recruited sequentially from a tertiary first-class hospital in Tianjin.The patients were assessed for bowel function using the LARS Score after anterior resection.Data were collected to conduct reliability tests on test-retest,inter-rater and constant,construction validity.Results The field test demonstrated excellent repeatability with an ICC value of 0.9615 (95％CI 0.9272～0.9796); inter-rater reliability was high with an ICC value of 0.9394 (95％CI 0.8854～0.9680).Content validity was excellent which CVR was 0.90.Constructive validity was good,factor analysis extracted two common factors,which could explain 60.659％ of the total variance,and each item on the corresponding factor had satisfactory factor loading quantity (＞0.4).Conclusions The Chinese version of LARS Score is easy to use and convenient to understand; the evidence collected in this study has shown good reliability and validity of using the LARS Score in assessing bowel function of Chinese rectal cancer patients.

Pdm09 virus outbreak occurred in Mainland China in May 2009, a few months later, the prevalence of seasonal H1N1(sH1N1) influenza virus that already circulated in human for tens of years began to decline and disappeared afterwards. To identify the reason for the rapid decline of sH1N1 in mainland China, we sequenced the HA1 of sH1N1 during 2006-2011, and then analyzed the selective pressure in different phases. Our results showed before Pdm09 outbreak, the omega value was 0. 36 while after Pdm09 outbreak the omega value was 0. 28 and significant difference (t test, P<0. 05) was identified. We concluded that sH1N1 obtained stronger purifying selection after Pdm09 outbreak in China. This might one of the major reasons causing the disappearance of sH1N1 in human.
China ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; virology ; Phylogeny ; Seasons ; Selection, Genetic