ObjectiveTo investigate whether Huanglian Jiedutang （HLJD） and its major active constituents （geniposide， baicalin， and berberine） can inhibit the inflammatory response of BV2 cells under lipopolysaccharide （LPS） stimulation via the high-mobility group protein B1 （HMGB1）/Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway， and to explore differences in therapeutic efficacy among the three monomers， their combined formula， and HLJD under equal content ratios. MethodsBV2 microglial cells were used as the primary experimental model. Cell viability was assessed using the cell counting kit-8 （CCK-8） method to examine the effects of different concentrations of dimethyl sulfoxide （DMSO， 0.8%， 0.4%， 0.2%， 0.1%， and 0.05%） on cell viability. IncuCyte was employed to monitor the growth of cells under different concentrations of HLJD （200， 100， 50， 25， 12.5， 6.25 mg·L^-1）. Nitric oxide （NO） assay was used to screen the optimal HLJD concentration. High-performance liquid chromatography （HPLC） determined the content of geniposide， baicalin， and berberine in HLJD， and experimental groups were subsequently established according to the relative proportions of these constituents. CCK-8 assay evaluated cell viability under different treatments. Enzyme-linked immunosorbent assay （ELISA） measured levels of inflammatory factors （TNF-α， IL-1β， IL-6， IL-10） in the supernatant. Flow cytometry assessed the effects of treatments on M1-type polarization of BV2 cells. Western blot determined the expression levels of HMGB1， TLR4， and NF-κB-related proteins. ResultsCompared with the blank group， DMSO at concentrations ≤0.2% did not affect cell viability within 48 h. BV2 cell growth plateaued at 24 h after treatment with 200 mg·L^-1 HLJD. Under stimulation with 2 mg·L^-1 LPS， this concentration of HLJD effectively reduced NO release， and 6 h pre-treatment had a stronger inhibitory effect on NO than direct administration. HPLC results showed that 1 mg of HLJD freeze-dried powder contained approximately 24 μg of geniposide， 15 μg of baicalin， and 30 μg of berberine. Based on these ratios， experimental groups were blank， LPS （2 mg·L^-1）， HLJD （200 mg·L^-1）， monomer combination， geniposide （4.8 mg·L^-1）， baicalin （3 mg·L^-1）， and berberine （6 mg·L^-1）. The monomer combination group consisted of all three active constituents dissolved together. LPS and HLJD or its active constituents did not affect cell viability compared with the blank group. LPS significantly increased TNF-α， IL-1β， IL-6， and IL-10 in the supernatant （P<0.01）. HLJD and its active constituents significantly reduced pro-inflammatory factors TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01） while upregulating anti-inflammatory IL-10 （P<0.01）， with the monomer combination showing the strongest effect （P<0.05， P<0.01）. Compared with the blank group， LPS significantly increased the proportion of CD80⁺CD86⁺ （M1-type） BV2 cells （P<0.01）. HLJD and its constituents partially inhibited M1 polarization （P<0.05， P<0.01）， with the monomer combination exhibiting the most pronounced effect （P<0.05， P<0.01）. Compared with the blank group， LPS upregulated HMGB1， TLR4， and NF-κB-related proteins （P<0.01）， whereas HLJD and its active constituents significantly reduced their expression （P<0.05， P<0.01）， with the monomer combination having the strongest regulatory effect （P<0.05， P<0.01）. ConclusionHLJD and its major active constituents （geniposide， baicalin， berberine） can inhibit LPS-induced inflammatory responses in BV2 cells. The combination of the three active constituents demonstrates the most potent anti-inflammatory effect， significantly attenuating M1-type polarization of BV2 cells via the HMGB1/TLR4/NF-κB signaling pathway.

Spine fracture and dislocation are common traumatic spinal conditions that often require surgical intervention due to compromised spinal stability. Surgical approaches include anterior, posterior, and combined anterior-posterior spinal procedures. According to the specific surgical requirements, patients may be placed in the prone position or repositioned between prone and supine positions during surgery. Intraoperative repositioning has become an essential step in patient positioning. However, during repositioning, patients with spinal fracture and dislocation are at increased risk for complications such as hemodynamic instability, nerve injury, and pressure injuries to the skin and soft tissue. Notably, due to the instability of the spinal cord, even minor manipulations can further exacerbate the damage, potentially leading to severe outcomes like paraplegia. Although the current clinical guidelines provide instructive recommendations for standard position, there remains no specific protocols for intraoperative repositioning in patients with spine fracture and dislocation. With a concern for the lack of clinical studies on positioning techniques, risk prevention, and operational norms for special patients, no applicable guidelines or standards are available. A consensus was required to provide clinical reference, meet the requirements of surgical treatment, and minimize the safety risks of patients caused by improper placement of positions. Professional Committee of Operating Room Nursing of Shaanxi Nursing Association organized experts in nursing management and operating room nursing from major hospitals across China to formulate Expert consensus on intraoperative repositioning for patients with spinal fracture and dislocation ( version 2025). The consensus provides 11 recommendations covering pre-repositioning preparation, intraoperative maneuvers, and post-repositioning observation, aiming to provide references for clinical standardization of the intraoperative repositioning process and protection of patients′ safety.

PURPOSE: Septic shock is associated with high mortality and poor outcomes among sepsis patients with coagulopathy. Although traditional statistical methods or machine learning (ML) algorithms have been proposed to predict septic shock, these potential approaches have never been systematically compared. The present work aimed to develop and compare models to predict septic shock among patients with sepsis. METHODS: It is a retrospective cohort study based on 484 patients with sepsis who were admitted to our intensive care units between May 2018 and November 2022. Patients from the 908th Hospital of Chinese PLA Logistical Support Force and Nanchang Hongdu Hospital of Traditional Chinese Medicine were respectively allocated to training (n=311) and validation (n=173) sets. All clinical and laboratory data of sepsis patients characterized by comprehensive coagulation indexes were collected. We developed 5 models based on ML algorithms and 1 model based on a traditional statistical method to predict septic shock in the training cohort. The performance of all models was assessed using the area under the receiver operating characteristic curve and calibration plots. Decision curve analysis was used to evaluate the net benefit of the models. The validation set was applied to verify the predictive accuracy of the models. This study also used Shapley additive explanations method to assess variable importance and explain the prediction made by a ML algorithm. RESULTS: Among all patients, 37.2% experienced septic shock. The characteristic curves of the 6 models ranged from 0.833 to 0.962 and 0.630 to 0.744 in the training and validation sets, respectively. The model with the best prediction performance was based on the support vector machine (SVM) algorithm, which was constructed by age, tissue plasminogen activator-inhibitor complex, prothrombin time, international normalized ratio, white blood cells, and platelet counts. The SVM model showed good calibration and discrimination and a greater net benefit in decision curve analysis. CONCLUSION The SVM algorithm may be superior to other ML and traditional statistical algorithms for predicting septic shock. Physicians can better understand the reliability of the predictive model by Shapley additive explanations value analysis.
Humans ; Shock, Septic/blood* ; Machine Learning ; Male ; Female ; Retrospective Studies ; Middle Aged ; Aged ; Sepsis/complications* ; ROC Curve ; Cohort Studies ; Adult ; Intensive Care Units ; Algorithms ; Blood Coagulation ; Critical Illness

Objective To investigate the effect and mechanism of actin gamma1(ACTG1)downregulation on apoptosis in gastric cancer cells line HGC-27.Methods Stable ACTG1 knockdown HGC-27 cell line was constructed by CRISPR/Cas9,ACTG1 knockdown was verified by DNA sequencing and Western blot,apoptosis rate was detected by flow cytometry,Western blot detected the expression level of apoptosis-related proteins Bcl2-associated X protein(Bax),B cell lymphoma 2 family protein(Bcl2)and PI3K/AKT signal pathway-related proteins AKT and p-AKT.Results HGC-27 cell lines with stable knockdown of ACTG1 were constructed,and the ACTG1 protein levels were decreased in the sgACTG1-1 and sgACTG1-2 groups compared with the Control group.Compared with the apoptosis rate in the Control group(6.54%±0.67%),cell apoptosis rate in the sgACTG1-1 and sgACTG1-2 groups(10.11%±0.46%,14.67%±0.17%)were significantly increased,the differences were statistically significant(t=-7.58,-20.28,all P<0.01).Compared the Control group,the Bax protein levels in the sgACTG1-1 and sgACTG1-2 groups(0.89±0.02,1.00±0.08 vs 0.71±0.03)were significantly increased(t=-8.14,-5.87),the level of Bcl2 protein(0.49±0.06,0.39±0.06 vs 0.65±0.07)were significantly decreased(t=3.09,5.35),the differences were statistically significant(all P<0.05).Compared with the Control group,the AKT protein levels in the sgACTG1-1 and sgACTG1-2 groups(0.95±0.10,0.43±0.09 vs 1.17±0.06)and the P-AKT protein level(0.38±0.08,0.28±0.12 vs 0.70±0.14)were significantly decreased,the differences were statistically significant(t=3.20,12.13;3.44,3.85,all P<0.05).Conclusion Downregulation of ACTG1 inhibits the expression of PI3K/Akt signaling pathway related proteins AKT,p-AKT and promotes gastric cancer cells line HGC-27 apoptosis.

Objective To investigate the effect and mechanism of actin gamma1(ACTG1)downregulation on apoptosis in gastric cancer cells line HGC-27.Methods Stable ACTG1 knockdown HGC-27 cell line was constructed by CRISPR/Cas9,ACTG1 knockdown was verified by DNA sequencing and Western blot,apoptosis rate was detected by flow cytometry,Western blot detected the expression level of apoptosis-related proteins Bcl2-associated X protein(Bax),B cell lymphoma 2 family protein(Bcl2)and PI3K/AKT signal pathway-related proteins AKT and p-AKT.Results HGC-27 cell lines with stable knockdown of ACTG1 were constructed,and the ACTG1 protein levels were decreased in the sgACTG1-1 and sgACTG1-2 groups compared with the Control group.Compared with the apoptosis rate in the Control group(6.54%±0.67%),cell apoptosis rate in the sgACTG1-1 and sgACTG1-2 groups(10.11%±0.46%,14.67%±0.17%)were significantly increased,the differences were statistically significant(t=-7.58,-20.28,all P<0.01).Compared the Control group,the Bax protein levels in the sgACTG1-1 and sgACTG1-2 groups(0.89±0.02,1.00±0.08 vs 0.71±0.03)were significantly increased(t=-8.14,-5.87),the level of Bcl2 protein(0.49±0.06,0.39±0.06 vs 0.65±0.07)were significantly decreased(t=3.09,5.35),the differences were statistically significant(all P<0.05).Compared with the Control group,the AKT protein levels in the sgACTG1-1 and sgACTG1-2 groups(0.95±0.10,0.43±0.09 vs 1.17±0.06)and the P-AKT protein level(0.38±0.08,0.28±0.12 vs 0.70±0.14)were significantly decreased,the differences were statistically significant(t=3.20,12.13;3.44,3.85,all P<0.05).Conclusion Downregulation of ACTG1 inhibits the expression of PI3K/Akt signaling pathway related proteins AKT,p-AKT and promotes gastric cancer cells line HGC-27 apoptosis.

Spine fracture and dislocation are common traumatic spinal conditions that often require surgical intervention due to compromised spinal stability. Surgical approaches include anterior, posterior, and combined anterior-posterior spinal procedures. According to the specific surgical requirements, patients may be placed in the prone position or repositioned between prone and supine positions during surgery. Intraoperative repositioning has become an essential step in patient positioning. However, during repositioning, patients with spinal fracture and dislocation are at increased risk for complications such as hemodynamic instability, nerve injury, and pressure injuries to the skin and soft tissue. Notably, due to the instability of the spinal cord, even minor manipulations can further exacerbate the damage, potentially leading to severe outcomes like paraplegia. Although the current clinical guidelines provide instructive recommendations for standard position, there remains no specific protocols for intraoperative repositioning in patients with spine fracture and dislocation. With a concern for the lack of clinical studies on positioning techniques, risk prevention, and operational norms for special patients, no applicable guidelines or standards are available. A consensus was required to provide clinical reference, meet the requirements of surgical treatment, and minimize the safety risks of patients caused by improper placement of positions. Professional Committee of Operating Room Nursing of Shaanxi Nursing Association organized experts in nursing management and operating room nursing from major hospitals across China to formulate Expert consensus on intraoperative repositioning for patients with spinal fracture and dislocation ( version 2025). The consensus provides 11 recommendations covering pre-repositioning preparation, intraoperative maneuvers, and post-repositioning observation, aiming to provide references for clinical standardization of the intraoperative repositioning process and protection of patients′ safety.

BACKGROUND: The effects of acupuncture have varied in different randomized controlled trials (RCTs), and there are many factors that influence treatment effect of acupuncture in different outcomes, with conflicting results. OBJECTIVE: To identify factors and their impact on the treatment effect of acupuncture in different outcomes. METHODS: Acupuncture RCTs were searched from 7 databases including Medline (PubMed), Embase, Cochrane Central Register of Controlled Trials, China National Knowledge Infrastructure, Wanfang Database, VIP Database, and China Biology Medicine disc between January 1st, 2015 and December 31st, 2019. Eligible studies must compare acupuncture to no acupuncture, sham acupuncture, or waiting lists, and report at least 1 patient-important outcome. A multi-level meta-regression was conducted using a 3-level robust mixed model and univariate analyses were performed for all independent variables, even those excluded from the multivariable model due to collinearities. We used thresholds of 0.2 and 0.4 for the difference of standardized mean differences (SMDs), categorising them as small (<0.2), moderate (0.2-0.4), or large (>0.4) effects. RESULTS: The pain construct analysis involved 211 effect estimates from 153 studies and 14 independent variables. High-frequency acupuncture treatment sessions produced larger effects compared to low-frequency sessions [large magnitude, the difference of adjusted SMDs 0.46, 95% confidence interval (CI) 0.07 to 0.84; P=0.02]. The non-pain symptoms construct analysis comprised 323 effect estimates from 231 studies and 15 independent variables. Penetrating acupuncture showed moderately larger effects when compared to non-penetrating acupuncture (0.30, 95% CI 0.06 to 0.53; P=0.01). The function construct analysis included 495 effect estimates from 274 studies and 14 independent variables. Penetrating acupuncture and the flexible acupuncture regimen showed moderately larger effects, compared to non-penetrating acupuncture and fixed regimen, respectively (0.40, 95% CI 0 to 0.80; P=0.05; 0.29, 95% CI 0.06 to 0.53; P=0.01). CONCLUSIONS High-frequency acupuncture sessions appear to be a more effective approach to managing painful symptoms. Penetrating acupuncture demonstrated greater effect in relieving non-painful symptoms. Both penetrating acupuncture type and flexible acupuncture regimen were linked to significant treatment effects in function outcomes. Future studies should consider the factors that are significantly associated with the effects of acupuncture in patient-important outcomes.
Humans ; Randomized Controlled Trials as Topic ; Acupuncture Therapy/methods* ; Pain ; Pain Management ; China

This study aims to investigate the mechanism of berberine in regulating the metabolism network via clock-controlled genes represented by brain and muscle arnt-like 1(BMAL1) to ameliorate insulin resistance(IR) of hepatocytes in vitro. The HepG2 cell model of dexamethasone-induced IR(IR-HepG2) was established and treated with 5, 10, and 20 μmol·L~(-1) berberine, respectively, for 24 h. The glucose oxidase method and cell counting kit-8(CCK-8) assay were employed to measure extracellular glucose concentration and cell viability, respectively. Periodic acid-Schiff(PAS) staining and lipid fluorescence method were used to detect glycogen and lipids. The immunofluorescence(IF) assay was employed to detect the nuclear localization of BMAL1 and circadian locomotor output cycles kaput(CLOCK) in IR-HepG2 cells. Western blot was employed to determine the protein levels of BMAL1, CLOCK, period circadian clock 2(PER2), cryptochrome circadian regulator 1(CRY1), Rev-Erbα, carbohydrate response element-binding protein(ChREBP), peroxisome proliferator-activated receptors alpha and gamma(PPARα/γ), sterol regulatory element-binding protein 1C(SREBP-1C), mammalian target of rapamycin(mTOR), protein kinase B(Akt), glycogen synthase kinase-3β(GSK3β), acetyl coenzyme A carboxylase 1(ACC1), fatty acid synthase(FASN), carnitine palmitoyltransferase 1α(CPT1α), nicotinamide phosphoribosyltransferase(NAMPT), silent information regulator 1(SIRT1), adiponectin(ADPN), insulin receptor substrate 2(IRS2), and phosphatidylinositol 3-kinase regulatory subunit p85(PI3Kp85). In addition, the levels of phosphorylated adenosine monophosphate-activated protein kinase alpha(AMPKα), Akt, GSK3β, BMAL1, and mTOR were determined. Furthermore, 20 μmol·L~(-1) CLK8 was added to measure the glucose consumption as well as the protein levels of ChREBP, PPARα, and mTOR in IR-HepG2 cells. The results showed that berberine increased the glucose consumption, lowered the lipid levels, increased the expression and nuclear localization of BMAL1 and CLOCK, and up-regulated the level of BMAL1 in IR-HepG2 cells. Furthermore, berberine up-regulated the levels of ADPN, IRS2, PI3Kp85, p-Akt(Ser473)/Akt, p-mTOR(Ser2448)/mTOR, PPARα, and CPT1α, and down-regulated the levels of p-GSK3β(Ser9)/GSK3β, ChREBP, SREBP-1C, ACC1, and FASN. The addition of CLK8 reduced glucose consumption in IR-HepG2 cells, up-regulated the ChREBP level, and down-regulated PPARα and mTOR levels by inhibiting the BMAL1 and CLOCK interaction. In summary, berberine regulated glucose and lipid metabolism via clock-controlled genes with BMAL1 at the core to ameliorate IR of hepatocytes.
Humans ; Hepatocytes/drug effects* ; Lipid Metabolism/drug effects* ; Glucose/metabolism* ; Berberine/pharmacology* ; Insulin Resistance ; Hep G2 Cells ; CLOCK Proteins/genetics* ; ARNTL Transcription Factors/genetics*

Objective To analyze the expression of serum microRNA(miR)-145 and miR-195 in patients with oral squamous cell carcinoma(OSCC)and their relationship with cervical lymph node metastasis.Methods A total of 102 patients diagnosed with OSCC in the hospital from January 2020 to June 2022 were selected as OSCC group.Clinicopathological parameters of OSCC patients were collected and sorted.According to whether cervical lymph node metastasis occurred,the patients were divided into a cervical lymph node me-tastasis group(43 cases)and a non-cervical lymph node metastasis group(59 cases).In the same period,79 patients with precancerous lesion were selected as the precancerous lesion group,and 86 healthy subjects with-out oral disease were selected as the healthy control group.Real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression levels of serum miR-145 and miR-195.The evaluation value of relative expression levels of serum miR-145 and miR-195 in cervical lymph node metastasis in OSCC patients was examined by receiver operating characteristic curve analysis.Results The relative expression lev-els of serum miR-145 and miR-195 in OSCC group were lower than those in precancerous lesion group and healthy control group,and the difference was statistically significant(P＜0.05).The relative expression levels of serum miR-145 and miR-195 in precancerous lesion group were lower than those in healthy control group,and the difference was statistically significant(P＜0.05).The relative expression levels of serum miR-145 and miR-195 in OSCC patients with low tumor differentiation and TNM stage Ⅲ+Ⅳ were lower than those in OSCC patients with medium/high tumor differentiation and TNM stage Ⅰ+Ⅱ,with statistical significance(P＜0.05).The relative expression levels of serum miR-145 and miR-195 in cervical lymph node metastasis group were lower than those in non-cervical lymph node metastasis group,and the difference was statistically significant(P＜0.05).The relative expression levels of serum miR-145 and miR-195 were 0.840(95％CI:0.765-0.915),0.832(95％CI:0.754-0.910)and 0.898(95％CI:0.754-0.910)of cervical lymph node metastasis in OSCC patients respectively.The specificity was 79.1％,88.4％and 77.5％,and the sensitivity was 74.6％,67.8％and 87.4％,respectively.Conclusion Serum miR-145 and miR-195 are low expressed in OSCC patients,and are related to cervical lymph node metastasis,tumor differentiation,and TNM stage in OSCC patients,which can be used as an important indicator to evaluate cervical lymph node metastasis in OS-CC patients.

Objective To explore the value of the Kyoto classification score in differentiating undifferentiated gastric cancer from differentiated gastric cancer,and establish a predictive scoring system for differentiating undifferentiated gastric cancer under endoscope.Methods 183 gastric cancer patients were retrospectively analyzed.According to pathology,95 patients were included in the differentiated group and 88 were included in the undifferentiated group.The age,gender and Kyoto classification score of patients in the two groups were compared,and the factors associated with undifferentiated gastric cancer were screened by binary Logistic regression analysis.The predictive scoring system for undifferentiated gastric cancer was established based on the obtained odds ratio(O(R))values,and the receiver operator characteristic curve(ROC curve)was drawn.Results Compared with differentiated group,the total scores of Kyoto classification,atrophy,intestinal metaplasia and diffuse redness were lower in undifferentiated group(P<0.01).Under the age of 55(P<0.05),female(P<0.05),and C1 atrophy or no atrophy(P<0.01)were independently associated with undifferentiated gastric cancer.The area under the curve(AUC)of predictive scoring system for undifferentiated gastric cancer was 0.881(95％CI:0.828～0.934),and the sensitivity and specificity were 80.70％and 90.50％at the optimal cut-off value.Conclusion There are differences in Kyoto classification scores between undifferentiated and differentiated gastric cancer patients.The predictive scoring system of undifferentiated gastric cancer established by us has certain value in distinguishing undifferentiated gastric cancer under endoscope.