1.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
2.Oxylipidomics Combined with Transcriptomics Reveals Mechanism of Jianpi Huogu Prescription in Treating Steroid-induced Osteonecrosis of Femoral Head in Rats
Lili WANG ; Qun LI ; Zhixing HU ; Qianqian YAN ; Liting XU ; Xiaoxiao WANG ; Chunyan ZHU ; Yanqiong ZHANG ; Weiheng CHEN ; Haijun HE ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):190-199
ObjectiveTo unveil the mechanism of Jianpi Huogu prescription (JPHGP) in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head (SONFH) by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal, model, low-, medium-, and high-dose (2.5, 5, 10 g·kg-1, respectively) JPHGP, and Jiangushengwan (1.53 g·kg-1) groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg-1 on days 1 and 2, and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection, and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head, and the number of adipocytes, the rate of empty bone lacunae, and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), apolipoprotein A1 (ApoA1), and apolipoprotein B (ApoB). At the same time, the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed, and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking, immunohistochemistry, and Western blot. ResultsCompared with the normal group, the model group showcased thinning of the femoral head, trabecular fracture, karyopyknosis, subchondral cystic degeneration, increases in the number of adipocytes and the rate of empty bone lacunae (P<0.01), a reduction in the trabecular area (P<0.01), decreases in BMD, Tb.Th, Tb.N, and BV/TV, and increases in Tb.Sp and BS/BV (P<0.01). Compared with the model group, the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae, an increase in the trabecular area (P<0.05, P<0.01), rises in BMD, Tb.Th, Tb.N, and BV/TV, and decreases in Tb.Sp and BS/BV (P<0.05, P<0.01). Compared with the normal group, the model group showcased raised serum levels of TG, TC, LDL, and ApoB and lowered serum levels of HDL and ApoA1 (P<0.01). Compared with the model group, the JPHGP groups had lowered serum levels of TG, TC, LDL, and ApoB (P<0.05, P<0.01) and a risen serum level of ApoA1 (P<0.05, P<0.01). Moreover, the serum level of HDL in the high-dose JPHGP group increased (P<0.01). A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head, and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation, lipids, apoptosis, and osteoclast differentiation. Molecular docking, immunohistochemistry, and Western blot results showed that compared with the normal group, the model group displayed increased content of 5-lipoxygenase (5-LO) and peroxisome proliferator-activated receptor γ (PPARγ) in the femoral head (P<0.01). Compared with the model group, medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ (P<0.05, P<0.01). ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.
3.The Effects of Tai Chi Training on Bone Density,Bone Turnover Markers,and Heart Rate Variability in High-Risk Osteoporosis Population
Jiaming LIN ; Chao LI ; Wei ZHAO ; Jun ZHOU ; Xiaoying CHEN ; Xiangyu XI ; Haijun HE ; Baohong MI ; Yuefeng CHEN ; Weiheng CHEN
Journal of Traditional Chinese Medicine 2025;66(15):1566-1571
ObjectiveTo explore the effects of the Tai Chi training on bone density, bone turnover markers, and heart rate variability for people with high-risk osteoporosis, and to provide evidence for the prevention of osteoporosis at early stage. MethodsSixty-six cases of people with high risk of osteoporosis were included, and they were divided into 33 cases each in the intervention group and the control group using the random number table method. The control group received osteoporosis health education three times a week, and the intervention group received Tai Chi training under the guidance of a trainer three times a week for 40 mins each time on the basis of the control group, and both groups were intervened for 12 weeks. Dual-energy X-ray absorptiometry was used to measure the bone density of L1~L4 vertebrae, bilateral femoral necks and bilateral total hips in the two groups before and after the intervention; enzyme-linked immunosorbent assay was used to determine bone turnover markers before and after the intervention, including pro-collagen type Ⅰ pro-amino-terminal prepropyl peptide (P1NP) and β-collagen type Ⅰ cross-linking carboxy-terminal peptide (β-CTX). Seven cases with good compliance in the intervention group were selected. After wearing the heart rate sensor, they successively performed Tai Chi training and walking activities recommended by the guideline for 20 mins each, and the heart rate variability (HRV) during exercise was collected, including time-domain indexes such as standard deviation of normal sinus intervals (SDNN), root-mean-square of the difference between adjacent RR intervals (RMSSD), frequency-domain metrics such as low-frequency power (LF), high-frequency power (HF), and low-frequency/high-frequency power ratio (LF/HF), as well as nonlinear metrics such as approximate entropy (ApEn), sample entropy (SampEn). ResultsFinally, 63 cases were included in the outcome analysis, including 30 cases in the intervention group and 33 cases in the control group. After the intervention, the differences of L1~L4 vertebrae, bone density of bilateral femoral neck and bilateral total hip in the intervention group were not statistically significant when compared with those before intervention (P>0.05), while the bone density of all parts of the control group decreased significantly compared with that before intervention (P<0.05), and the difference in the bone density of the L1~L4 vertebrae, bilateral femoral neck, and the right total hip before and after the intervention of the intervention group was smaller than that of the control group (P<0.05). The differences in P1NP and β-CTX between groups before and after intervention was not statistically significant (P>0.05). Compared with walking exercise, LF decreased, HF increased and LF/HF decreased during Tai Chi exercise (P<0.05); the time domain indexes and non-linear indexes between groups had no statistically significant difference (P>0.05). ConclusionTai Chi exercise can maintain lumbar, hip, and femoral bone density and improve sympathetic/parasympathetic balance in people at high risk for osteoporosis, but cannot significantly improve bone turnover markers.
4.Level of coagulation factor Xin patients with venous thrombosis of lower limbs and its correlation with recurrence risk
Haijun JIANG ; Ronghua LI ; Guoping CHEN ; Li WANG ; Jun HOU ; Xiaokang CHENG ; Liming YU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(2):151-154
Objective To investigate the level of coagulation factor Ⅺ(FⅪ)in patients with venous thrombosis of lower limbs and its correlation with recurrence risk.Methods A total of 220 pa-tients with deep vein thrombosis(DVT)admitted in our hospital from February 2018 to February 2019 were enrolled as the study group,and another 50 healthy individuals taking physical exami-nation during same period served as the control group.After a 3 years followed,the study group ultimately included 197 cases,according to the results of restricted cubic spline(RCS),the study group was divided into low(FⅪ<10.3 U/L,94 cases),medium-(10.3-12.1 U/L,52 cases)and high-level groups(>12.1 U/L,51 cases).The plasma level of FⅪ was detected in the study group 1 month after the end of anticoagulant therapy,and the results were compared with those of the control group during physical examination.Cox model was used to analyze the influence of FⅪ on the recurrence of DVT,and RCS was employed to analyze the relationship between DVT recur-rence and FⅪ level.Kaplan-Meier curve was plotted to analyze the recurrence risk of DVT with different FⅪ levels.The patients from the study group were followed up for 3 years.Results The FⅪ level was significantly higher in the study group than the control group(P<0.05).During fol-low-up period,33 patients(16.75%)had DVT recurrence.The Cox model analysis after adjust-ment of sex and age showed that FⅪ level was a risk factor for DVT recurrence(P<0.05).When the FⅪ level was set into tertile and the risk ratio was calculated after adjustment,FⅪ<10.3 U/L,and the average FⅪ level at this stage was 9.2 U/L,the risk ratio was 0.82(95%CI:0.673-0.984);Patients with FⅪ between 10.3 and 12.1 U/L,and the average FⅪ at this stage was 11.4 U/L,the risk ratio of 1.04(95%CI:0.813-1.432).The those with FⅪ>12.1 U/L,and the average FⅪ at this stage was 13.8 U/L,hazard ratio of 1.38(95%CI:0.921-1.563).Kaplan-Meier curve analysis showed that the recurrence risk was 28.62%(95%CI:25.633-31.609),30.10%(95%CI:27.594-32.606)and 38.06%(95%CI:34.306-41.371),respectively for the low-,medium-,and high-level groups,with significant correlation among the three groups(x2=6.631,P=0.036).Conclusion Compared with healthy individuals,plasma FⅪ level is at a high level in the DVT patients.With the increment of FⅪ level,the risk of DVT recurrence increases.Two FⅪ levels,10.3 U/L and 12.1 U/L,can be used as reference points for the obvious increase of DVT recur-rence rate.
5.Prevalence of cataracts and the coverage rate of cataract surgery in Ningxia region
Jinjin ZHANG ; Lin CHEN ; Tian TIAN ; Haijun LIU ; Wei NIU ; Xue ZHANG ; Mengli JI ; Wenjuan ZHUANG
Chinese Journal of Experimental Ophthalmology 2024;42(3):279-284
Objective:To investigate the prevalence of cataracts, the surgical coverage, and postoperative visual acuity of adults in Ningxia.Methods:A cross-sectional study using multistage cluster random sampling was conducted.Ten survey sites in Ningxia were selected and the population aged 18 years and over was surveyed with questionnaire, height and weight measurements, visual acuity, intraocular pressure, fundus photography and slit-lamp examinations.Cataract prevalence and its influencing factors were analyzed.Cataract prevalence, surgical coverage and presenting visual acuity (PVA) and best corrected visual acuity (BCVA) after surgery were investigated in different age groups of the examined population.The study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of the People's Hospital of Ningxia Hui Autonomous Region (No.[2023]-LL-010).Participants signed informed consent prior to the examination.Results:A total of 6 145 people should be examined, and 5 721 people were actually examined, with an examination rate of 93.10%.The study population consisted of 2 558 males, accounting for 44.71%, and 3 163 females, accounting for 55.28%, with ages ranging from 18 to 93 years old and an average age of (64.27±13.48) years.Among them, 1 180 patients diagnosed with cataract, with a cataract prevalence of 20.62%.The prevalence of cataract increased with age and decreased with education level, showing statistically significant differences ( χ2=1 091.32, 581.92; both at P<0.01).The prevalence of cataract was significantly higher among people with hypertension, diabetes mellitus, hyperlipidemia, and coronary heart disease than those without these diseases ( χ2=274.65, 118.15, 78.05, 182.71; all at P<0.01).Cataract surgery was performed in 245 cases in the cataract patient population, with a surgical coverage rate of 20.76%.Of the 245 cases, 229 cases were implanted with IOLs, with an implantation rate of 93.40%.The social burden rate of cataract blindness was 2.29%, and increased with age.Of the 339 eyes that underwent cataract surgery, 241 had a PVA≥0.3, accounting for 71.09%, and 272 had a BCVA≥0.3, accounting for 80.24%. Conclusions:In Ningxia, cataracts are still the main cause of vision impairment and blindness in the elderly, and the social burden rate of cataract blindness is high.Moreover, the coverage rate of cataract surgery is low, so both the coverage and quality of surgery need improvement.
6.Curcumin protects against liver cancer in a mouse model by down-regu-lating HO-1/NQO1
Haijun MOU ; Xingxing CHEN ; An'an LIU ; Li ZHANG ; Jiaxin ZHU ; Hai JIN
Chinese Journal of Pathophysiology 2024;40(3):438-443
AIM:To observe the effect of curcumin on a C57BL/6J mouse liver cancer model induced by N-ni-trosodiethylamine(DEN)combined with carbon tetrachloride(CCl4),and to explore its mechanism.METHODS:Forty young male C57BL/6J mice were intraperitoneally injected with DEN(25 mg/kg)14 d after birth and randomly divided in-to the following 4 groups at the 4th week(10 in each group):model control group and curcumin(100,200 and 400 mg/kg)groups.Ten male mice of the same age were used as normal control group.The mice in model group and curcumin groups were gavaged with 10%CCl4(5 mL/kg)twice a week from the 8th week on.At the same time,the mice in curcumin groups were gavaged with curcumin,and the mice in normal control group were gavaged with the same volume of distilled water once a day for 14 weeks.After administration,the mice were sacrificed,the liver surface was observed,and the number of tumor nodules was compared.The activity of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum was detected by an automatic biochemical instrument.The pathological changes of liver tissues were ob-served by HE staining.The mRNA expression levels of heme oxygenase-1(HO-1)and NAD(P)H-quinone oxidoreductase 1(NQO1)were detected by RT-qPCR,and the protein expression levels of HO-1,NQO1 and Ki67 were detected by Western blot and immunohistochemistry.RESULTS:Compared with normal control group,the body weight of the mice in model group was decreased significantly(P<0.01),the liver index was increased significantly(P<0.01),and the se-rum levels of ALT and AST were increased obviously(P<0.01).There was no significant difference in the mRNA expres-sion levels of HO-1 and NQO1(P>0.05),the protein levels of HO-1 and NQO1 were increased distinctly(P<0.05),and the positive expression rate of Ki67 was increased significantly(P<0.05).After curcumin treatment,the body weight of the mice was significantly increased(P<0.01),the liver index was not changed(P>0.05),and the number of tumor nodules in the liver was decreased significantly(P<0.05 or P<0.01).The serum level of AST was decreased significantly(P<0.01),the mRNA and protein expression levels of HO-1 and NQO1 were decreased significantly(P<0.05),and the posi-tive expression rate of Ki67 was decreased significantly(P<0.05).CONCLUSION:Curcumin significantly protects against liver cancer in a C57BL/6J mouse model induced by DEN combined with CCl4,and its mechanism may be related to the inhibition of HO-1 and NQO1 expression.
7.Effects of higher femoral tunnels on clinical outcomes, MRI, and second-look findings in double-bundle anterior cruciate ligament reconstruction with a minimal 5-year follow-up
Lin LIN ; Haijun WANG ; Jian WANG ; Yongjian WANG ; Yourong CHEN ; Jiakuo YU
Chinese Medical Journal 2024;137(4):465-472
Background::To perform anatomical anterior cruciate ligament reconstruction (ACLR), tunnels should be placed relatively higher in the femoral anterior cruciate ligament (ACL) footprint based on the findings of direct and indirect femoral insertion. But the clinical results of higher femoral tunnels (HFT) in double-bundle ACLR (DB-ACLR) remain unclear. The purpose was to investigate the clinical results of HFT and lower femoral tunnels (LFT) in DB-ACLR.Methods::From September 2014 to February 2016, 83 patients who underwent DB-ACLR and met the inclusion and exclusion criteria were divided into HFT-ACLR (group 1, n = 37) and LFT-ACLR (group 2, n = 46) according to the position of femoral tunnels. Preoperatively and at the final follow-up, clinical scores were evaluated with International Knee Documentation Committee (IKDC), Tegner activity, and Lysholm score. The stability of the knee was evaluated with KT-2000, Lachman test, and pivot-shift test. Cartilage degeneration grades of the International Cartilage Repair Society (ICRS) were evaluated on magnetic resonance imaging (MRI). Graft tension, continuity, and synovialization were evaluated by second-look arthroscopy. Return-to-sports was assessed at the final follow-up. Results::Significantly better improvement were found for KT-2000, Lachman test, and pivot-shift test postoperatively in group 1 ( P >0.05). Posterolateral bundles (PL) showed significantly better results in second-look arthroscopy regarding graft tension, continuity, and synovialization ( P <0.05), but not in anteromedial bundles in group 1. At the final follow-up, cartilage worsening was observed in groups 1 and 2, but it did not reach a stastistically significant difference ( P >0.05). No statistically significant differences were found in IKDC subjective score, Tegner activity, and Lysholm score between the two groups. Higher return-to-sports rate was found in group 1 with 86.8% (32/37) vs. 65.2% (30/46) in group 2 ( P = 0.027). Conclusion::The HFT-ACLR group showed better stability results, better PL, and higher return-to-sports rate compared to the LFT-ACLR group.
8.Effect of human bone marrow mesenchymal stem cells on biological behavior of human liposarcoma SW872 cells through YAP
Hua CHEN ; Na SHA ; Ning LIU ; Yang LI ; Haijun HU
Journal of Jilin University(Medicine Edition) 2024;50(4):1000-1008
Objective:To observe the effect of human mesenchymal stem cells(hMSCs)conditioned medium(CM)co-cultured with the human liposarcoma SW872 cells on the proliferation and migration of the tumor cells,and to discuss the effect of hMSCs CM on the liposarcoma cells and the possible mechanism.Methods:The hMSCs were cultured in vitro and transfected with either lentiviral vector control shNS(control group)or lentiviral shRNA targeting Yes-associated protein(YAP)(shYAP-hMSCs group)by lentiviral methods.The expression levels of YAP mRNA and protein in the hMSCs in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.The CM was then harvested.The SW872 cells were cultured in vitro and divided into control group(normal culture),hMSCs CM group,and shYAP-hMSCs CM group.The proliferation activities of the cells in various groups were detected by CCK-8 assay;the apoptotic rates of the cells in various groups were detected by flow cytometry;the scratch healing rates of the cells in various groups were detected by cell scratch assay;the expression levels of YAP,matrix metallopeptidase-9(MMP-9),and cyclin D1 proteins in the cells in various groups were detected by Western blotting method.Results:Compared with control group,the expression levels of YAP mRNA and protein in the cells in shYAP-hMSCs group were decreased(P<0.01),indicating the successful establishment of a stable transfected cell line.The CCK-8 assay results showed that compared with control group,the proliferation activity of the cells in hMSCs CM group was increased(P<0.05),and the proliferation activity of the cells in shYAP-hMSCs CM group was decreased(P<0.01).The flow cytometry results showed that compared with control group,there was no significant change in the apoptotic rate of the cells in hMSCs CM group(P>0.05),while the apoptotic rate of the cells in shYAP-hMSCs CM group was increased(P<0.01).The cell scratch assay results showed that compared with control group,the scratch healing rate of the cells in hMSCs CM group was increased(P<0.05),and the scratch healing rate of the cells in shYAP-hMSCs CM group was decreased(P<0.01).The Western blotting results showed that compared with control group,there were no significant differences in the expression levels of YAP,MMP-9,and cyclin D1 proteins in the cells in hMSCs CM group(P>0.05),while the expression levels of YAP,MMP-9,and cyclin D1 proteins in the cells in shYAP-hMSCs group were decreased(P<0.05 or P<0.01).Conclusion:The hMSCs regulate the proliferation and migration of the human liposarcoma SW872 cells,and its mechanism may be related to the expression of YAP.
9.Porphyromonas gingivalis promotes the occurrence of esophageal squamous cell carcinoma via an inflammatory microenvironment
Haijun XU ; Yijun QI ; Dangrou WU ; Qiwei LIU ; Pan CHEN ; Mengxiang LI ; Yelin JIAO ; Haojie RUAN ; Zhitao LI ; Shegan GAO
Chinese Journal of Oncology 2024;46(8):746-754
Objective:To investigate the role of an inflammatory microenvironment induced by Porphyromonasgingivalis ( P. gingivalis) in the occurrence of esophageal squamous cell carcinoma (ESCC) in mice. Methods:A total of 180 C57BL/6 mice were randomly divided into 6 groups, i.e. control group, P. gingivalis group, 4NQO group, 4NQO + P. gingivalis group, 4NQO + P. gingivalis + celecoxib group, and 4NQO + P. gingivalis + antibiotic cocktail (ABC, including metronidazole, neomycin, ampicillin, and vancomycin) group, with 30 mice in each group, using the random number table. All mice were normalized by treatment with ABC in drinking water for 2 weeks. In the following 2 weeks, the mice in the control group and the P. gingivalis group were given drinking water, while the other 4 groups were treated with 30 μg/ml 4NQO in the drinking water. In weeks 11-12, the mice in the P. gingivalis group, the 4NQO + P. gingivalis group, the 4NQO + P. gingivalis + celecoxib group, and the 4NQO + P. gingivalis + ABC group were subjected to ligation of the second molar in oral cavity followed by oral P. gingivalis infection thrice weekly for 24 weeks in weeks 11-34. In weeks 13-34, the mice in 4NQO + P. gingivalis+celecoxib group and 4NQO + P. gingivalis + ABC group were administered with celecoxib and ABC for 22 weeks, respectively. At the end of 34 weeks, gross and microscopic alterations were examined followed by RT-qPCR and immunohistochemistry to examine the expression profiles of inflammatory- and tumor-molecules in esophagi of mice. Results:At 34 weeks, 4NQO treatment alone did not affect the foci of papillary hyperproliferation, diseased area, and the thickness of the esophageal wall, but significantly enhanced the foci of hyperproliferation (median 1.00, P<0.05) and mild/moderate dysplasia (median 2.00, P<0.01). In addition, the expression levels of IL-6 [8.35(3.45,8.99)], IL-1β [6.90(2.01,9.72)], TNF-α [12.04(3.31,14.08)], c-myc [2.21(1.80,3.04)], pSTAT3, Ki-67, and pH2AX were higher than those in the control group. The pathological changes of the esophageal mucosa were significantly more overt in the 4NQO + P. gingivalis group in terms of the foci of papillary hyperproliferation (median 2.00), diseased area (median 2.51 mm 2), the thickness of the esophageal wall (median 172.52 μm), the foci of hyperproliferation (median 1.00, P<0.05), and mild/moderate dysplasia (median 1.00, P<0.01). In mice of the 4NQO + P. gingivalis group, the expression levels of IL-6 [12.27(5.35,22.08)], IL-1β [13.89(10.04,15.96)], TNF-α [19.56(6.07,20.36)], IFN-γ [11.37(8.23,20.07)], c-myc [2.62(1.51,4.25)], cyclin D1 [4.52(2.68,7.83)], nuclear pSTAT3, COX-2, Ki-67, and pH2AX were significantly increased compared with the mice in the control group. In mice of the 4NQO + P. gingivalis group, the diseased area, invasive malignant foci as well as pSTAT3 and pH2AX expression were significantly blunted by celecoxib. Treatment with ABC markedly reduced the papillary hyperproliferative foci, invasive malignant foci, and pSTAT3 expression but not pH2AX. Conclusions:P. gingivalis promotes the occurrence of esophageal squamous cell carcinoma in mice by inducing an inflammatory microenvironment primed with 4NQO induced DNA damage. Clearance of P. gingivalis with ABC or anti-inflammatory intervention holds promise for prevention of esophageal squamous cell malignant pathogenesis via blockage of IL-6/STAT3 signaling and amelioration of inflammation.
10.Effects of propofol on blood pressure and heart rate in hypertensive patients with hyperacute uncomplicated type B aortic dissection
Wuhong ZHENG ; Peng XIAO ; Haijun ZHOU ; Zongcun HE ; Feng CHEN ; Jun KE
Chinese Journal of Emergency Medicine 2024;33(7):968-974
Objective:To investigate the clinical efficacy of propofol on blood pressure (BP) and heart rate (HR) in hypertensive patients with hyperacute uncomplicated type B aortic dissection (HU-TBAD).Methods:This study was a single-center, double-blind, randomized controlled trial. Totally 96 consecutive hypertensive patients with HU-TBAD admitted to the Department of Emergency in our hospital from July 2020 to March 2023 were enrolled and randomly divided into control and treatment groups ( n=48/group) by envelope method. All patients were treated with nicardipine, remifentanil, esmolol, and basic treatments. Besides, patients in the treatment group were injected with 0.5 mg/kg propofol, followed by 1.0 mg/(kg·h) with continuous micro-pump intravenous infusion; the RASS score was evaluated every 15 minutes to adjust the dosage of propofol to maintain the RASS score at -2-0 points, while the control group was given an equal volume of normal saline. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and HR were analyzed at different time points (TPs). Related indexes between the two groups were compared at 0 (T 0 min) and 60 (T 60 min) minutes. Standard-reaching rate of related indexes, levels of mean nicardipine dose (mND) and urine volume, and adverse effect rates (AERs) were also compared between the two groups. All patients were admitted to the cardiovascular surgical ward to receive proper management and follow-up for 21 d after discharge from the Department of Emergency. Mann-Whitney U test, t-test, χ2 test, or Fisher's test were used to compare the data between the two groups, while the data of two groups at different TPs were compared by using repetitive measurement deviation analysis. Results:No significant differences were observed in general clinical data between the two groups (all P>0.05). There were significant differences in SBP, DBP, and HR levels in different TPs, groups, and interaction of Time and Group (Time×Group) (all P<0.05). For comparison of related indexes at T 0 min and T 60 min, there were statistical differences in oxygenation index levels at different TPs ( P<0.01), but not in different groups and Time×Group (all P>0.05); significant differences in levels of partial pressure of carbon dioxide, respiratory rate, and lactate were observed in different TPs and Time×Group (all P<0.01), but not observed in different groups (all P>0.05). There were significant differences in NRS score in different TPs, groups, and Time×Group (all P<0.05), while not in cardiac troponin I levels in different TPs, groups, and Time×Group (all P>0.05). Compared with the control group, the standard-reaching rate of SBP, DBP, HR, sedation, and analgesia as well as the level of RASS score reduction in the treatment group were significantly increased [54.17% vs. 77.08%, 56.25% vs. 81.25%, 50.00% vs. 72.92%, 47.92% vs. 72.92%, 43.75% vs. 83.33%, 1.00 (0, 2.00) vs. 2.00 (1.00, 3.00), respectively, all P<0.05], while the level of mND was significantly decreased [μg·kg -1·min -1, 2.50 (2.00, 2.50) vs. 2.00 (1.50, 2.50), P<0.01]; there were no statistical differences in both urine volume levels and AERs between the two groups (all P>0.05). Following up for 21 d, the rate of aortic dissection deterioration and ICU admission was significantly lower in the treatment group than in the control group (19.57% vs. 4.26%, 23.91% vs.6.38%, respectively, all P<0.05). Conclusions:Propofol enhances the analgesic effect of remifentanil, synergistically reduces SBP, DBP, and HR, and improves clinical prognosis in hypertensive patients with HU-TBAD.

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