[Objective] To extract hemoglobin (Hb) from red blood cells using osmotic pressure swelling method, expected to achieve a hemoglobin dissolution rate of ≥80% and a cell membrane integrity rate of ≥70%. [Methods] Human umbilical cord blood red blood cells were used as raw materials and phosphate buffer solution was used as the swelling solution for red blood cells. A three factor three-level orthogonal experiment (n=3) was conducted to determine the optimal matching conditions for selecting the osmolality molar concentration of phosphate buffer solution, pH value of hypotonic phosphate buffer solution and volume ratio of hypotonic phosphate buffer solution to washed red blood cells. Red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions. The hemoglobin dissolution rate and cell membrane integrity rate were checked. In the expanded comparative experiment, red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions, which was filtered by ultrafiltration membranes. The filtration time and hemoglobin yield were checked. [Results] The optimal matching conditions for preparing red blood cell swelling solution were obtained through orthogonal experiment as follows: osmotic pressure molar concentration was 30 mOsmol/Kg, pH was 7.8, and phosphate buffer to red blood cell volume ratio was 6∶1. On the basis of the above conditions, the red blood cell swelling solution sample was compared with the original process sample: the hemoglobin dissolution rate was (82.4±1.8)% vs (78.6±3.0)% (P<0.05), and the cell membrane integrity rate was (65.8±4.0)% vs (28.7±2.3)% (P<0.05). In the expanded comparative experiment, the optimal matching conditions were compared with the original process conditions: filtration time(s) (327±9) vs (434±13) (P<0.05), and hemoglobin yield was (72.3±1.2)% vs (66.0±1.4)% (P<0.05). [Conclusion] Compared with the original preparation process, the hemoglobin extraction process which optimized through orthogonal experiments greatly reduces the cell membrane fragmentation rate and minimizes the entry of cell membrane matrix into the target solution, ensuring a slightly higher hemoglobin dissolution rate, and reducing the preparation difficulty for the subsequent cell membrane separation and further purification.

[Objective] To investigate the protective effect of Salvia miltiorrhiza injection (SMI) on the kidneys of HBOC-CHP01 resuscitated haemorrhagic shock rats. [Methods] A 50% haemorrhagic shock rat model was established, with 12 rats divided into two groups: SMI + HBOC-CHP01 group and HBOC-CHP01 group, with 6 rats in each group. The rats in the SMI+ HBOC-CHP01 group were given an equal volume of HBOC-CHP01 for resuscitation after haemorrhagic shock, and an 8 mL/kg dose of SMI. Rats in the HBOC-CHP01 group were resuscitated by administering an equilibrium blood loss volume of HBOC-CHP01 and given an 8 mL/kg dose of 0.9% NaCl solution. Blood was taken from rats at five points: before bloodletting (baseline), during haemorrhagic shock (HS), immediately after resuscitation (RS0h), 1 h after resuscitation (RS1h), and 24 h after resuscitation (RS24h). A blood gas analyser was used to detect the lactate level (Lac), glucose content (Glu), residual base (BEecf), pH, bicarbonate (HCO3-), high iron haemoglobin (MetHb). White blood cells (WBC), platelets (PLT), haemoglobin content (Hb), carboxyhaemoglobin (COHb) were detected using a quintuple classification. Blood creatinine (SCr), uric acid (UA), kidney-related indexes were detected using biochemistry instrument. Kidney tissues of the rats were taken after 24 h of resuscitation and after execution, and the inflammation of kidneys of the rats of the two groups was analyzed using HE staining. Fluorescence staining was used to detect the level of ROS in the kidneys of rats in both groups. [Results] At RS 0h, the Beecf, Glu and Lac levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group, and the pH level of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group, and the Glu levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group at RS 1h. At RS 0h, the WBC, PLT and COHb contents of rats in the SMI+HBOC-CHP01 group were all significantly higher than those of rats in the HBOC-CHP01 group, and at RS 1h, the WBC content of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group; at RS 1h, the UA content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the SCr content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the inflammation level of kidney tissues of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC -CHP01 group rats, and the ROS and MPO levels in the kidney tissues of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group. [Conclusion] The combination of Salvia miltiorrhiza injection during the resuscitation of rats with severe haemorrhagic shock by HBOC-CHP01 can alleviate renal injury by reducing inflammatory response and oxidative stress.

Objective:To explore the mechanism of Coicis Semen in treating non -small cell lung cancer combined with network pharmacology and in vitro cell experiment.Methods:The components and potential targets of Coicis Semen were obtained from TCMSP, and NSCLC related targets were collected from Genecards and Drugbank. They were intersected to get common targets. An intersection target PPI network was constructed using STRING database. GO function and KEGG pathway enrichment analysis were performed using DAVID database. A "disease-drug component-target" network was constructed using Cytoscape 3.8.0. The CCK-8 method was used to screen the concentration of Coicis Semen, and the cells were randomly divided into control group, Coicis Semen low-concentration group (0.125 mg/ml), Coicis Semen high-concentration group (0.25 mg/ml), Coicis Semen low concentration+FoxO1 inhibitor group, and FoxO1 inhibitor group (50 μmol/L) using a random number table. Cell proliferation ability was determined using CCK-8 method, and cell migration and invasion ability were detected using cell scratch assay and Transwell cell invasion assay. Western blot was used to detect the expression of FoxO1, cofilin-1, and p-cofilin-1 proteins.Results:A total of 9 active components and 246 targets were screened from Coicis Semen, 1 074 targets related to non-small cell lung cancer, and 11 intersecting targets were identified. The key targets of Coicis Semen in the treatment of non-small cell lung cancer were MAPK1, MAPK8, MAPK14, CDK2, INSR, AR, FGFR2, PGR, DHFR, and PLAU; GO functional enrichment analysis showed that Coicis Semen had an impact on the cellular, molecular, and biological functions of A549 cells; KEGG pathway enrichment analysis showed that the pathways of Coicis Semen in treating non-small cell lung cancer mainly focus on FoxO pathway, MAPK pathway, prostate cancer pathway, Ras pathway, etc. Compared with the control group, the proliferation rate, migration rate, and invasion number of A549 cells in the low and high concentration groups of Coicis Semen decreased ( P<0.05), p-cofilin-1 expression decreased ( P<0.05), and FoxO1 expression increased ( P<0.05). Conclusion:Coicis Semen can treat non-small cell lung cancer by regulating MAPK1, MAPK8, and MAPK14 targets, and acting on pathways such as the FoxO and MAPK pathways.

Objective To identify the risk factors and investigate etiological spectrum of pulmonary fungal infections(PFIs)in patients with end-stage liver disease(ESLD).Methods A retrospective analysis was performed on the clinical data of 211 ESLD patients.Based on pulmonary imaging,clinical manifestations,and microbiological test results,patients were categorized into three groups,including the PFI group(or the case group),the non-fungal pneumonia group(or the control group 1),and the group without pneumonia(or the control group 2).The clinical characteristics of patients in the the case group were then compared with those of patients in the two control groups.Taking patients without pneumonia as the control,univariate and multivariate logistic regression analyses were performed to identify independent risk factors for PFI,and a nomogram prediction model was constructed based on these risk factors.Results Among the 211 patients,76(36.1%)had PFIs,46(21.8%)had non-fungal pneumonia,and 89(42.2%)did not have pneumonia.According to findings from the multivariate logistic regression,elevated white blood cell count upon admission(OR=1.211;95%CI,1.011-1.460),higher Model for End-Stage Liver Disease-Sodium(MELD-Na)score(OR=1.140;95%CI,1.021-1.282),concomitant hepatorenal syndrome(OR=4.150;95%CI,1.050-17.300),cumulative glucocorticoid use for more than seven days(OR=26.832;95%CI,6.361-113.221),and the administration of broad-spectrum antibiotics at the time of hospital admission(OR=6.601;95%CI,1.951-22.362)were identified as independent risk factors for PFI.A predictive nomogram model named TJLFPFI was constructed based on these risk factors.The area under the receiver operating characteristic(AUC)curve of the model was 0.899(95%CI,0.853-0.945).Etiologic analysis of the 76 PFI cases revealed that 36(47.4%)had positive results for culture,while 40(52.6%)had negative results for sputum culture but tested positive by the 1,3-β-D-glucan test and/or galactomannan test.Aspergillus was the most frequently identified pathogen,detected in 25 of the 36 cases(59.5%).Conclusion PFI in ESLD patients is closely associated with disease severity at admission,early use of broad-spectrum antibiotics,and prolonged glucocorticoid therapy.Aspergillus is the predominant pathogen.The TJLFPFI model shows potential value in identifying high-risk patients,but prospective validation is still warranted.

Objective To explore the relationship between plasma trimethylamine N-oxide(TMAO)level and atrial fibrillation(AF)by a clinical trial.Methods Totally 257 patients with AF were enrolled including 114 patients with paroxysmal AF and 143 with persistent AF.The control group had 122 non-AF patients;Plasma TMAO level,the demographic characteristics and clinical data of patients were collected.Regression analysis was performed to identify the association between TMAO and presence of AF.Results Plasma TMAO level of patients with AF was higher than that of patients without AF(3.9[2.4-5.9]vs.2.4[1.6-3.7]μmol/L,P＜0.001);Even after adjust-ment of confounding factors including age,hypertension,diabetes,heart failure and body mass index;Plasma TMAO level was still related to presence of AF(OR:1.290,95％CI:1.151-1.445;P＜0.001).The plasma TMAO level of patients with persistent AF was higher than that of patients with paroxysmal AF(4.4[2.8-6.8]vs.3.2[2.2-5.0]μmol/L,P＜0.01);Even after adjustment of confounding factors including age,hypertension,diabetes and heart failure and coronary heart disease;Plasma TMAO level was still related to persistent AF(OR:1.092,95％CI:1.009-1.182;P=0.029).Conclusions The level of plasma TMAO of patients with AF is higher than that of patients without AF.Elevated plasma TMAO level is related to an increased risk of AF.Elevated plasma TMAO level in patients with AF is related to an increased risk of persistent AF.

BACKGROUND: The role of inflammation in the development of gestational diabetes mellitus (GDM) has recently become a focus of research. The systemic immune-inflammation index (SII) and systemic inflammation response index (SIRI), novel indices, reflect the body's chronic immune-inflammatory state. This study aimed to investigate the associations between the SII or SIRI and GDM. METHODS: A prospective birth cohort study was conducted at Beijing Obstetrics and Gynecology Hospital from February 2018 to December 2020, recruiting participants in their first trimester of pregnancy. Baseline SII and SIRI values were derived from routine clinical blood results, calculated as follows: SII = neutrophil (Neut) count × platelet (PLT) count/lymphocyte (Lymph) count, SIRI = Neut count × monocyte (Mono) count/Lymph count, with participants being grouped by quartiles of their SII or SIRI values. Participants were followed up for GDM with a 75-g, 2-h oral glucose tolerance test (OGTT) at 24-28 weeks of gestation using the glucose thresholds of the International Association of Diabetes and Pregnancy Study Groups (IADPSG). Logistic regression was used to analyze the odds ratios (ORs) (95% confidence intervals [CIs]) for the the associations between SII, SIRI, and the risk of GDM. RESULTS: Among the 28,124 women included in the study, the average age was 31.8 ± 3.8 years, and 15.76% (4432/28,124) developed GDM. Higher SII and SIRI quartiles were correlated with increased GDM rates, with rates ranging from 12.26% (862/7031) in the lowest quartile to 20.10% (1413/7031) in the highest quartile for the SII ( Ptrend <0.001) and 11.92-19.31% for the SIRI ( Ptrend <0.001). The ORs (95% CIs) of the second, third, and fourth SII quartiles were 1.09 (0.98-1.21), 1.21 (1.09-1.34), and 1.39 (1.26-1.54), respectively. The SIRI findings paralleled the SII outcomes. For the second through fourth quartiles, the ORs (95% CIs) were 1.24 (1.12-1.38), 1.41 (1.27-1.57), and 1.64 (1.48-1.82), respectively. These associations were maintained in subgroup and sensitivity analyses. CONCLUSION The SII and SIRI are potential independent risk factors contributing to the onset of GDM.
Humans ; Female ; Pregnancy ; Diabetes, Gestational/immunology* ; Prospective Studies ; Adult ; Inflammation/immunology* ; Glucose Tolerance Test ; Birth Cohort

OBJECTIVE: To investigate the effectiveness of one-stage direct suture technique under arthroscopy for the treatment of anterior cruciate ligament (ACL) injury combined with anterior meniscus root injury. METHODS: The clinical data of 9 patients with ACL injury or tibial intercondylar eminence fracture combined with anterior meniscus root injury who were admitted between September 2017 and September 2024 and met the selection criteria were retrospectively analyzed. There were 3 males and 6 females, aged 21-62 years, with an average age of 37.1 years. The time from injury to surgery ranged from 5 days to 5 years, with a median time of 40 days. Among them, 5 cases had ACL injury, including 4 cases of type 1 and 1 case of type 2 according to modified Sherman classification; 4 cases had tibial intercondylar eminence fracture, including 3 cases of type 3 and 1 case of type 2 according to Meyers-McKeever classification. There were 7 cases of anterior root injury of lateral meniscus and 2 cases of anterior root injury of medial meniscus. The preoperative International Knee Documentation Committee (IKDC) score was 45.0±12.3, and Lysholm score was 49.2±12.4. Preoperatively, 7 cases were positive in anterior drawer test, Lachman test, and McMurray test, while 2 cases could not complete the test due to pain limitation. Preoperatively and at last follow-up, IKDC score and Lysholm score were used to evaluate knee joint function, anterior drawer test and Lachman test were used to evaluate knee joint stability, and McMurray test was used to evaluate meniscus condition. RESULTS: The operation time was 30-100 minutes, with an average of 64.2 minutes; the total hospital stay was 2-12 days, with an average of 4.5 days; the postoperative hospital stay was 1-4 days, with an average of 1.8 days. All incisions healed by first intention without surgery-related complications. All 9 patients were followed up 2-30 months, with an average of 18.8 months. No internal fixation-related complications occurred during follow-up. At last follow-up, MRI review showed good ligament tension, and CT showed good fracture healing. The results of anterior drawer test and Lachman test were all negative. McMurray test was negative in all cases. The IKDC score was 88.3±5.1, and Lysholm score was 88.3±5.6, both showing significant improvement compared to preoperative scores ( t=14.001, P<0.001; t=10.192, P<0.001). CONCLUSION One-stage direct suture technique under arthroscopy for repairing ACL injury or tibial intercondylar eminence fracture combined with anterior meniscus root injury can achieve good effectiveness without fixation device-related complications.
Humans ; Adult ; Male ; Female ; Arthroscopy/methods* ; Middle Aged ; Anterior Cruciate Ligament Injuries/surgery* ; Tibial Meniscus Injuries/surgery* ; Retrospective Studies ; Suture Techniques ; Young Adult ; Treatment Outcome ; Tibial Fractures/surgery* ; Anterior Cruciate Ligament Reconstruction/methods* ; Menisci, Tibial/surgery*

Objective: To study the effect of nano hemoglobin-based oxygen carrier (nano-HBOC) on radiosensitivity of lung cancer H385 cells. Methods: Using 95% N and 5% CO , a lung cancer cell line was constructed in a hypoxic environment, and H385 cells were treated with different concentrations of nano-HBOC and irradiated (4Gy) by an irradiator, and the IC50 concentration was calculated. The cells were detected by flow cytometry (reactive oxygen species, ROS) ROS test. Using GEO database, KEGG pathway enrichment analysis was carried out to predict possible pathways. The levels of lipid peroxidation and Fe were observed by fluorescence microscope, and the proteins related to iron death pathway were detected by Western-blot. Results: Compared with the control cells, the activity and density of the cells were significantly decreased by nano-HBOC combined with radiotherapy, with a notable proportion of cells exhibiting deteriorated status. There is a positive correlation between ROS level and nano-HBOC concentration, especially after radiotherapy. Radiotherapy combined with nano-HBOC significantly increased the levels of lipid peroxidation and Fe in H385 cells, while decreasing the levels of iron death pathway proteins slc7a11 and GPX4, and increasing the level of ACSL4. Conclusion: Nano-HBOC enhances the radiosensitivity of lung cancer H385 cells.

Objective: To study the effect of nano hemoglobin-based oxygen carrier (nano-HBOC) on radiosensitivity of lung cancer H385 cells. Methods: Using 95% N and 5% CO , a lung cancer cell line was constructed in a hypoxic environment, and H385 cells were treated with different concentrations of nano-HBOC and irradiated (4Gy) by an irradiator, and the IC50 concentration was calculated. The cells were detected by flow cytometry (reactive oxygen species, ROS) ROS test. Using GEO database, KEGG pathway enrichment analysis was carried out to predict possible pathways. The levels of lipid peroxidation and Fe were observed by fluorescence microscope, and the proteins related to iron death pathway were detected by Western-blot. Results: Compared with the control cells, the activity and density of the cells were significantly decreased by nano-HBOC combined with radiotherapy, with a notable proportion of cells exhibiting deteriorated status. There is a positive correlation between ROS level and nano-HBOC concentration, especially after radiotherapy. Radiotherapy combined with nano-HBOC significantly increased the levels of lipid peroxidation and Fe in H385 cells, while decreasing the levels of iron death pathway proteins slc7a11 and GPX4, and increasing the level of ACSL4. Conclusion: Nano-HBOC enhances the radiosensitivity of lung cancer H385 cells.

Laboratory animals are the "living" tools of medical research. Through animal experiments, people can gain continuous insights into the laws of life, reveal the essence of diseases, develop vaccines and drugs for prevention and treatment, and play an important role in the technological development of fields related to human health. The environmental conditions of laboratory animals have a direct impact on their health, quality, and the results of animal experiments. The higher the degree of environmental control, the more reliable the experimental results are in terms of quality. Therefore, environmental control of laboratory animal facilities is important for ensuring that laboratory animals live under required conditions, which is a key factor for conducting effective animal experiments. This article analyzes the current status of environmental testing of laboratory animal facilities in Sichuan Province, briefly summarizing their number, area, and other basic information, and provides detailed statistics on the ability of institutions to conduct environmental testing for laboratory animal facilities in Sichuan Province. It also summarizes the testing requirements for laboratory animal facility environments based on national standards, regulatory requirements, and the quality control needs of facility users. In the analysis of testing indicators for laboratory animal facilities, based on testing data from 40 laboratory animal facilities in Sichuan Province, it was found that static pressure difference is the indicator most prone to non-compliance, followed by illumination and air exchange rate. Using barrier environments as examples, common problems in the process of environmental testing for laboratory animal facilities are summarized in six aspects: testing personnel, instruments, methods, technical materials, testing environment, and reports, and targeted suggestions are proposed. These suggestions help improve environmental control in laboratory animal facilities, and provide practical reference and guidance for relevant testing institutions, as well as laboratory animal producers and users in the industry.