Exploratory study of the influence of respiratory microbiology on the efficacy of PD-1 inhibitors monotherapy for patients with advanced non-small cell lung cancer / 中国临床药理学与治疗学

AIM: To investigate the implication of respiratory microbiology on the efficacy of PD-1 inhibitors monotherapy for patients with NSCLC. METHODS: This study was designed as a retrospective study, fifty-eight patients with previously-treated advanced NSCLC who were received PD-1 monotherapy from October 2018 to October 2021 were included. The PD-1 inhibitors were consisted of camrelizumab, sintilimab and pembrolizumab. Additionally, the basic demographic data, therapeutic efficacy data, survival prognosis and adverse reactions during the PD-1 inhibitors treatment were collected and analyzed through the patients' medical records of the department and the electronic medical record system of the hospital. Furthermore, deep induced sputum specimens of the patients before treatment with PD-1 inhibitor were collected. And the respiratory microbiology of 58 samples were detected using 16S rRNA gene sequencing method. The index of respiratory microbiology α diversity was analyzed, and the correlation analysis was performed with the efficacy and prognosis of patients. RESULTS: A total of 58 patients with advanced NSCLC met the study's screening criteria and were evaluable for efficacy and safety profile. Efficacy data suggested that the objective response rate (ORR) and disease control rate (DCR) of the patients who received PD-1 inhibitors was 19.0%(95% CI: 9.9%-31.4%) and 55.2% (95% CI: 41.5%-68.3%). Furthermore, prognostic data obtained from follow-up indicated that the median PFS of the 58 patients with advanced NSCLC was 3.2 months (95% CI: 2.29-4.11) and the median OS was 10.5 months (95% CI: 5.58-15.43). Regarding the exploratory analysis between efficacy and respiratory microbiology, the 58 patients with NSCLC were divided into high α diversity group (group H) and low α diversity group (group L) according to Shannon diversity index of respiratory microecology detection. And the association analysis suggested that the ORR of patients with group H and group L was 23.3% and 17.9%(P c 0.380), respectively. Furthermore, prognostic analysis indicated that the median PFS of patients with group H and group L was 3.8 and 2.8 months, respectively, which was statistically significant (P c 0.034). CONCLUSION: PD-1 inhibitors monotherapy demonstrated preliminary efficacy and prognosis as subsequent line treatment for patients with advanced NSCLC. Patients with higher α -diversity of respiratory microbiology might confer a superior prognosis. And the conclusion should be validated in large sample prospective clinical trials subsequently.

Diterpenoids of Tripterygium wilfordii decrease inflammatory response of macrophage by inhibiting multiple signaling pathways / 中国药理学通报

Aim To study the anti-inflammatory activ¬ity of diterpenes from Tripterygium wilfordii on lipopo- lysaccharide ( LPS)-induced macrophage and its mech¬anism. Methods MTT assay was used to detect the cytotoxicity of compounds. The Griess method was used to detect the NO on LPS-induced RAW264. 7 cells. ELISA was applied to determine the contents of inter- leukin 6 (IL-6) , tumor necrosis factor a ( TNF-a ) , interleukin lp (IL-lfj) and interleukin 18 (IL-18) in cell culture supernatant. Western blot was used to de¬tect IkBcx, .INK, ERK, p38, STAT3 and their phos-phorylation in LPS-induced RAW264.7, as well as the effect on COX-2, iNOS, NLRP3, caspase-1 , cleaved- caspase-1. Flow cytometry was employed to detect the effects of compounds on the phagocytosis of RAW 264. 7 cells. Results Hypoglicin II (1) and ent-pimara-8 (14) , 15-diene-19-ol (6) , two diterpenoid compounds from Tripterygium wilfordii could effectively inhibit the expression of inflammatory mediators ( COX-2 and iN- OS) and inflammatory cytokines (IL-6, IL-lp, IL- 18) in LPS-induced RAW264. 7 cells. Further re¬search found that the phosphorylation of IkBcx , JNK, ERK, P38, STAT3 and NLRP3 was all inhibited; however, there was no significant effect on the expres¬sion of IkBcx, JNK, ERK, P38 and STAT3. At the same time, they also inhibited the phagocytosis of mac-rophages. Conclusions The anti-inflammatory mecha¬nism of Tripterygium wilfordii diterpenoids 1 and 6 might be through inhibiting the production of NLRP3 inflammatory bodies, inflammatory mediators (COX-2 and iNOS) and inflammatory cytokines (IL-6, IL-lp and IL-18) , which is closely related to inhibiting the activation of MAPK, NF-kB and STAT3 pathway.

Tea and tea drinking: China's outstanding contributions to the mankind.

BACKGROUND: Tea trees originated in southwest China 60 million or 70 million years ago. Written records show that Chinese ancestors had begun drinking tea over 3000 years ago. Nowadays, with the aging of populations worldwide and more people suffering from non-communicable diseases or poor health, tea beverages have become an inexpensive and fine complementary and alternative medicine (CAM) therapy. At present, there are 3 billion people who like to drink tea in the world, but few of them actually understand tea, especially on its development process and the spiritual and cultural connotations. METHODS: We searched PubMed, Google Scholar, Web of Science, CNKI, and other relevant platforms with the key word "tea", and reviewed and analyzed tea-related literatures and pictures in the past 40 years about tea's history, culture, customs, experimental studies, and markets. RESULTS: China is the hometown of tea, tea trees, tea drinking, and tea culture. China has the oldest wild and planted tea trees in the world, fossil of a tea leaf from 35,400,000 years ago, and abundant tea-related literatures and art works. Moreover, tea may be the first Chinese herbal medicine (CHM) used by Chinese people in ancient times. Tea drinking has many benefits to our physical health via its antioxidant, anti-inflammatory, immuno-regulatory, anticancer, cardiovascular-protective, anti-diabetic, and anti-obesity activities. At the moment, COVID-19 is wreaking havoc across the globe and causing severe damages to people's health and lives. Tea has anti-COVID-19 functions via the enhancement of the innate immune response and inhibition of viral growth. Besides, drinking tea can allow people to acquire a peaceful, relaxed, refreshed and cheerful enjoyment, and even longevity. According to the meridian theory of traditional Chinese medicine, different kinds of tea can activate different meridian systems in the human body. At present, black tea (fermented tea) and green tea (non-fermented tea) are the most popular in the world. Black tea accounts for over 90% of all teas sold in western countries. The world's top-grade black teas include Qi Men black in China, Darjeeling and Assam black tea in India, and Uva black tea in Sri Lanka. However, all top ten famous green teas in the world are produced in China, and Xi Hu Long Jing tea is the most famous among all green teas. More than 700 different kinds of components and 27 mineral elements can be found in tea. Tea polyphenols and theaflavin/thearubigins are considered to be the major bioactive components of black tea and green tea, respectively. Overly strong or overheated tea liquid should be avoided when drinking tea. CONCLUSIONS: Today, CAM provides an array of treatment modalities for the health promotion in both developed and developing countries all over the world. Tea drinking, a simple herb-based CAM therapy, has become a popular man-made non-alcoholic beverage widely consumed worldwide, and it can improve the growth of economy as well. Tea can improve our physical and mental health and promote the harmonious development of society through its chemical and cultural elements.

Clinical characteristics of acute kidney injury in cancer patients receiving immune checkpoint inhibitors / 中华肾脏病杂志

Objective:To investigate the incidence, risk factors, and outcomes of acute kidney injury (AKI) in cancer patients receiving immune checkpoint inhibitors (ICIs).Methods:A retrospective analysis was performed on the inpatients who received ICIs therapy in Beijing Shijitan Hospital, Capital Medical University from October 2015 to December 2020. According to the Kidney Disease: Improving Global Outcomes (KDIGO) definition of AKI, patients were divided into AKI group and non-AKI group, and the patients in the AKI group were further divided into ICIs related AKI (ICIs-AKI) and AKI due to other etiologies. The clinical characteristics of the patients were compared. Multivariate logistic regression was used to analyze the influencing factors of AKI, and sensitivity analysis was used to evaluate the influencing factors of ICIs-AKI.Results:A total of 279 cancer patients over 18 years old were included in this study, in which 175(62.7%) were males. There were 41 patients (14.70%) in AKI group, including 25 patients (8.96%) in ICIs-AKI group and 16 patients (5.73%) in AKI due to other etiologies group. Patients in the AKI group were characterized by higher proportions of hypertension, diuretics use and baseline eGFR<60 ml·min -1·(1.73 m 2) -1, extrarenal immune-related adverse events (irAEs) and a lower plasma albumin level (all P<0.05). The patients in the ICIs-AKI group had higher proportions of new aseptic leukocyturia, blood eosinophil count>500/ml, combined extrarenal irAEs, glucocorticoid use and discontinued ICIs treatment (all P<0.05). Multivariate logistic regression results showed that hypertension ( OR=3.424, 95% CI 1.559-7.522, P=0.002), use of diuretics ( OR=4.620, 95% CI 2.111-10.112, P<0.001), baseline eGFR<60 ml·min -1·(1.73 m 2) -1 ( OR=3.668, 95% CI 1.336-10.070, P=0.012) and extrarenal irAEs ( OR=9.909, 95% CI 4.198-23.391, P<0.001) were associated with AKI in cancer patients receiving ICIs therapy. Sensitivity analysis indicated that the risk factors of ICIs-AKI included use of diuretics and baseline eGFR<60 ml·min -1·(1.73 m 2) -1, similar to the results of the above analysis, extrarenal irAEs ( OR=17.572, 95% CI 6.302-48.995, P<0.001) were also associated with ICIs-AKI independently. Conclusions:AKI is not uncommon in patients treated with ICIs. Concomitant hypertension, baseline eGFR<60 ml·min -1·(1.73 m 2) -1 and use of diuretics are independent risk factors for AKI in such patients. Patients should be alert to the risk of ICIs-AKI when appearing extrarenal irAEs. Distinguishing ICIs-AKI from AKI caused by other causes will present a frequent challenge to clinical practitioners.

Hamartoma of the breast in a man: A rare case report.

RATIONALE: Mammary hamartoma is a rare benign breast tumor, composed of ducts, lobules, fibers, and adipose tissue. We describe a mammary hamartoma in a man; this is the fourth case being reported in the literature. PATIENT CONCERNS: A 30-year-old man presented with a 1-month history of a painless mass in his right breast. DIAGNOSIS: Ultrasound imaging and mammography revealed a lesion, approximately 2.0 cm × 2.0 cm in size, in the right breast, which was considered to be either a lipomyoma or an adenoma fibrosum. INTERVENTIONS: The mass was surgically resected. Pathological examination confirmed the diagnosis of mammary hamartoma. OUTCOMES: The patient was discharged from the hospital after surgery. There was no sign of reoccurrence during a 1-year follow-up period. LESSONS: At present, mammary hamartoma is considered to be a benign lesion, usually treated by surgical resection. Some reports have suggested a possible association between a hamartoma and the development of breast malignancy. The pathology and biology of an association between a mammary hamartoma and malignancy have not been defined to date.

Observation on curative effects of neoadjuvant chemotherapy for different subtypes of Luminal B breast cancer and analysis on its prognosis / 吉林大学学报(医学版)

Objective:To investigate the differences in the curative effects of neoadjuvant chemotherapy(NAC) for different subtypes of Luminal B breast cancer and its prognosis,and to discuss the clinical treatment characteristics of different subgroups.Methods:A total of 246 cases of Luminal B like breast cancer patients who completed the projected NAC courses and surgical treatment were selected.All the biopsy specimens before treatment were positive for estrogen receptot(ER).According to the expressions of progesterone receptor(PR), human epidermal growth factor-2(Her-2)and cell proliferation nuclear antigen Ki-67,246 cases of Luminal B breast cancer patients were divided into 3 subgroups.A subgroup(PR low expression group),Her-2 negative and PR< 20% or negative,Ki-67 any levels;B subgroup(PR high expression group),Her-2 negative,PR≥20% and Ki-67 ≥14%;C subgroup(Her-2 positive group),Her-2 positive,Ki-67 and PR any levels.The clinical pathological materials and follow-up recurrence events of the patients were collected.Results:Among the three subtypes of Luminal B breast cancer,there were no significant differences in the age,the size of primary tumor and the stage of TNM(P>0.05).There were significant differences in the lymph node metastasis rate among three subgroups(P=0.018),and the lymph node metastasis rate was the highest in A subgroup among three subgroups. There were no significant differences in the clinical response and pathological response among three subgroups(P=0.123,P=0.06).8.5%(21/246)patients achieved the pathological complete response(pCR);the rates of pCR in three subgroups had statistically significant difference(P=0.009);the rate of pCR in C subgroup was the highest, and the rate of pCR in B subgroup was the lowest.The Log-Rank test of the survival curves of three subgroups had not statistically significant difference(P=0.216),but the 3-year disease-free survival(DFS)and 5-year DFS of the patients in B subgroup were slightly higher than those in other two groups.The DFS of the patients in C subgroup was longer than that of the patients with Her-2 overexpression breast cancer at the same period,and the difference was statistically significant(P=0.047).Conclusion:Her-2 positive Luminal B breast cancer is more likely to achieve pCR in NAC and the prognosis is better than Her-2 overexpressing breast cancer.The patients with high expression of PR in Luminal B breast cancer patients have a tend of overall survival advantage compared with the patients with PR low expression and Her-2 positive expression.

Tribulus terrestris L protects the photoreceptors against light-induced degeneration in part through suppression of retinal inflammation and oxidative stress / 眼科新进展

Objective To investigate the protective effects of tribulus terrestris L (TTL) against light-induced photoreceptor degeneration and its underlying mechanisms.Methods BALB/c mice were randomly divided into normal control group,light exposure group and TTL administration group,with 12 mice in each group,and then exposed to light at the intensity of 10,000 lux for 30 min for the establishment of a retinal damage model of BALB/C mice.And 100 μL TTL decoction was intraperitoneally administered into mice of TTL administration group 30 min prior to illumination.Saline vehicle was administrated into the mice of normal control group and light exposure group.Next,intraperitoneal injection of dihydroethidium (DHE) was performed 22 h after illumination,and the eyes were enucleated 2 h later and subjected to cryosectioning for microscopic detection of the in situ retinal oxidative stress.Then,retinas were dissected 6 and 24 h after illumination,which was followed by total RNA extraction,reverse transcription and RT-PCR to assess the expression level of proinflammatory cytokines.Meanwhile,the expression levels of interleukin-β (IL-1 β),chemokine (Ccl2),cyclooxygenase-2 (COX-2),tumor necrosis factor-α (TNF-α),cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) mRNA were determined.Results Prominent oxidative stress was observed in retinal pigment epithelial cells and photoreceptor cells in the light exposure group when compared with the normal control group and TTL administration group,with significant difference (both P ＜ 0.001).Moreover,results of RT-PCR revealed that the expression of IL-1β,Ccl2,COX-2,TNF-α,ICAM-1 and VCAM-1 mRNA was significantly elevated as a result of light exposure compared to those from vehicle-treated normal controls with a significant difference (all P ＜ 0.01).TTL treatment resulted in significantly decreased expression of IL-1β,Ccl2,COX-2,TNF-α,ICAM-1 and VCAM-1 mRNA compared to those from light-exposed vehicle-treated controls with significant differences (all P ＜0.01).Conclusion In the retinal degeneration model,TTL protects the photoreceptor cells against fight-induced degeneration in part through suppressing light-induced retinal oxidative stress and inflammatory responses.

Trend Analyses of Users of a Syringe Exchange Program in Philadelphia, Pennsylvania: 1999-2014.

This study examines trends of injection drug users' (IDUs) use of a Philadelphia, Pennsylvania, syringe exchange program (SEP) from 1999 to 2014, including changes in demographics, drug use, substance abuse treatment, geographic indicators, and SEP use. Prevention Point Philadelphia's SEP registration data were analyzed using linear regression, Pearson's Chi square, and t-tests. Over time new SEP registrants have become younger, more racially diverse, and geographically more concentrated in specific areas of the city, corresponding to urban demographic shifts. The number of new registrants per year has decreased, however syringes exchanged have increased. Gentrification, cultural norms, and changes in risk perception are believed to have contributed to the changes in SEP registration. Demographic changes indicate outreach strategies for IDUs may need adjusting to address unique barriers for younger, more racially diverse users. Implications for SEPs are discussed, including policy and continued ability to address current public health threats.

Elimination of disinfection byproduct formation potential in reclaimed water during solar light irradiation.

Ecological storage of reclaimed water in ponds and lakes is widely applied in water reuse. During reclaimed water storage, solar light can degrade pollutants and improve water quality. This study investigated the effects of solar light irradiation on the disinfection byproduct formation potential in reclaimed water, including haloacetonitriles (HANs), trichloronitromethane (TCNM), trihalomethanes (THMs), haloketones (HKs) and chloral hydrate (CH). Natural solar light significantly decreased the formation potential of HANs, TCNM, and HKs in reclaimed water, but had a limited effect on the formation potential of THMs and CH. Ultraviolet (UV) light in solar radiation played a dominant role in the decrease of the formation potential of HANs, TCNM and HKs. Among the disinfection byproducts, the removal kinetic constant of dichloroacetonitrile (DCAN) with irradiation dose was much larger than those for dichloropropanone (1,1-DCP), trichloropropanone (1,1,1-TCP) and TCNM. During solar irradiation, fluorescence spectra intensities of reclaimed water also decreased significantly. The removal of tyrosine (Tyr)-like and tryptophan (Trp)-like protein fluorescence spectra intensity volumes was correlated to the decrease in DCAN formation potential. Solar irradiation was demonstrated to degrade Trp, Tyr and their DCAN formation potential. The photolysis products of Trp after solar irradiation were detected as kynurenine and tryptamine, which had chloroform, CH and DCAN formation potential lower than those of Trp.

Optimum target plasma concentration of propofol in preventing adverse effects of carboprost tromethamine in patients undergoing caesarean section / 中华麻醉学杂志

Objective To investigate the optimum target plasma concentration of propofol in preventing the adverse effects of carboprost tromethamine in the patients undergoing caesarean section.Methods One hundred and twenty-eight nulliparous parturients who were at full term with a singleton fetus,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 24-37 yr,weighing 54-78kg,scheduled for elective caesarean section under combined spinal-epidural anesthesia,were randomly divided into 4 groups (n =32 each) using a random number table:control group (group C),and different concentrations of propofol groups (P1-3 groups).Carboprost tromethamine 250 μg was injected into the body of the uterus,and propofol with the target plasma concentrations of 0.8,1.2 and 1.6 μg/ml was simultaneously given by target-controlled infusion in P1,P2 and P3 groups,respectively,and normal saline was infused at a rate of 0.5 ml · kg-1 · h-1 in group C.The occurrence of cardiovascular events was recorded from the end of carboprost tromethamine administration until the end of surgery.The relatedadverse effects after carboprost tromethamine administration,and Ramsay sedation score at 15 mm after carboprost tromethamine administration were recorded,and satisfactory sedation was defined as Ramsay sedation score of 2.The occurrence of complications associated with combined spinal-epidural anesthesia was recorded during the postoperative follow-up.Results Compared with group C,the incidence of carboprost tromethamine-related adverse effects was significantly decreased in P2 and P3 groups,the rate of satisfactory sedation was significantly increased in P1 and P2 groups,the incidence of hypotension and tachycardia was significantly decreased in group P1 (P＜0.05),and no significant change was found in the incidence of carboprost tromethamine-related adv erse effects in group P1,and in the rate of satisfactory sedation in group P3 (P＞ 0.05).Compared with group P1,the incidence of carboprost tromethaminerelated adverse effects was significantly decreased in P2 and P3 groups,the rate of satisfactory sedation was significantly increased in group P2,and the rate of satisfactory sedation was significantly decreased in group P3 (P＜0.05).Compared with group P2,the rate of satisfactory sedation was significantly decreased (P＜0.05),and no significant change was found in the incidence of carboprost tromethamine-related adverse effects in group P3 (P＞0.05).No cardiovascular events were found in group P2,and the incidence of hypotension was 9％ in group P3.Conclusion The optimum target plasma concentration of propofol in preventing the adverse effects of carboprost tromethamine is 1.2 μg/ml in the patients undergoing caesarean section.

Lipoxin A4 Activates Nrf2 Pathway and Ameliorates Cell Damage in Cultured Cortical Astrocytes Exposed to Oxygen-Glucose Deprivation/Reperfusion Insults.

Lipoxin A4 (LXA4), a potent antioxidant and anti-inflammation mediator, protects brains against cerebral ischemia/reperfusion (I/R) injury in vivo. However, few reports concern its function on astrocytes during cerebral I/R injury. The pathogenesis of cerebral I/R injury involves oxidative stress caused by reactive oxygen species (ROS). Upregulation of nuclear factor erythroid 2-related factor 2 (Nrf2) is generally considered to reduce oxidative stress. Nrf2 can induce heme oxygenase-1 (HO-1) expression and glutathione (GSH) release to combat increased oxidative stress. We investigated the effects of LXA4 on astrocytic cell damage, the production of ROS, and Nrf2 pathway, especially on HO-1 expression and GSH release in cultured cortical astrocytes exposed to oxygen-glucose deprivation (OGD)/recovery (OGDR) insults. Primary astrocytes were subjected to a 4-h OGD, followed by 8-h recovery. Cell viability, the production of ROS, and GSH release were measured. Furthermore, Nrf2, HO-1, and p62 expression levels were determined by Western blot. Moreover, Nrf2 location was studied by immunofluorescence staining. Treatment of LXA4 attenuates OGDR-induced cell damage and the production of ROS in a concentration-related manner. LXA4 induced Nrf2 expression and its nuclear translocation, as well as HO-1 expression and GSH release. Moreover, LXA4 induced the excess p62 accumulation. These results indicate that LXA4 can effectively protect against OGDR-induced cell damage in astrocytes, and activation of Nrf2 pathway to reduce oxidative stress may be involved in its protective effects. p62 accumulation induced by LXA4 may be closely related to Nrf2 activation.

Analysis on related factors of axillary lymph node metastasis in patients with cT1-2 N0 breast cancer / 吉林大学学报(医学版)

Objective To investigate the relationships between axillary lymph node metastasis and clinicopathologic features in the patients with cT1-2 N0 breast cancer and clarify the law of axillary lymph node metastasis,and to find the risk factor,and provide the theoretical basis for individuation therapy.Methods 687 patients with cT1-2 N0 breast cancer were divided into negative group and positive group according to the pathological results of axillary lymph node,and the clinicopathologic features were layered.The risk factors of axillary lymph node metastasis were screened out by Chi-square test and Logistic regression analysis.Results In 687 cases of cT1-2 N0 breast cancer,156 (22.7%)cases were observed with axillary lymph node metastasis. The age,cT stage,pT stage, pathological type,vascular invasion,perineural invasion estrogen receptor (ER),progesterone receptor (PR), and molecular subtyping were the factors that influenced axillary lymph node metastasis in univariate analyses.The age < 35 years, cT2 , invasive ductal carcinoma, vascular invasion positive and Luminal subtyping were the independent risk factors of axillary lymph nodes metastasis in multivariate analyses (r = 3.440,P = 0.010;r =1.770,P =0.007;r = 3.397,P = 0.001;r = 7.434,P = 0.000;r = 2.212,P = 0.015).Conclusion In the patients with cT1-2 N0 breast cancer,the age,cT,pathological type,vascular invasion and molecular subtyping are important predictors of axillary lymph node metastasis and vascular invasion was the most important predictor.The preoperative comprehensive analysis and evaluation of clinical data and preoperative pathological results obtained will help to select the right surgical operation.

Lipoxin A4 ameliorates cerebral ischaemia/reperfusion injury through upregulation of nuclear factor erythroid 2-related factor 2.

OBJECTIVES: Lipoxin A4 (LXA4) is a potent anti-inflammatory mediator that exerts a neuroprotective effect following cerebral ischaemia/reperfusion (I/R) injury. However, little is known about the underlying mechanisms. Upregulation of nuclear factor erythroid 2-related factor 2 (Nrf2) is generally considered to reduce cerebral I/R injury. Nuclear factor erythroid 2-related factor 2 can induce haeme oxygenase-1 (HO-1) and glutathione (GSH) expression to combat increased oxidative stress. The present study aimed to investigate the effects of Nrf2 signalling on LXA4-mediated neuroprotection. METHODS: Adult male Sprague Dawley rats were subjected to 2-hour middle cerebral artery occlusion followed by 24-hour reperfusion. Rats were randomly divided into four groups: Sham, I/R, LXA4, and LXA4+butoxycarbonyl-Phe-Leu-Phe-Leu-Phe (Boc2) (all n = 24). Brain infarction was detected by 2,3,5-triphenyltetrazolium chloride staining. After 24 hours of reperfusion, Nrf2, HO-1, and p62 expression levels were determined by western blot, and GSH synthesis was assessed. RESULTS: Lipoxin A4 effectively reduced infarct volumes and improved neurological scores. These effects were partially blocked by Boc2, a specific antagonist of the LXA4 receptor (ALXR). Lipoxin A4 induced Nrf2 expression and its nuclear translocation, as well as HO-1 expression and GSH synthesis; Boc2 did not block these effects. The excess p62 accumulation induced by LXA4 might be closely related to Nrf2 activation. DISCUSSION: Overall, our data suggest that Nrf2 upregulation is involved in the neuroprotective effects of LXA4 and may be ALXR independent.

Lipoxin A4 inhibits 5-lipoxygenase translocation and leukotrienes biosynthesis to exert a neuroprotective effect in cerebral ischemia/reperfusion injury.

Lipoxin A(4) (LXA(4)), a biologically active eicosanoid with anti-inflammatory and pro-resolution properties, was recently found to have neuroprotective effects in brain ischemia. As 5-lipoxygenase (5-LOX) and leukotrienes are generally considered to aggravate cerebral ischemia/reperfusion (I/R) injury, we investigated their effects on LXA(4)-mediated neuroprotection by studying middle cerebral artery occlusion (MCAO)/reperfusion in rats and oxygen-glucose deprivation (OGD)/recovery in neonatal rat astrocyte primary cultures. LXA(4) effectively reduced infarct volumes and brain edema, and improved neurological scores in the MCAO/reperfusion experiments; this effect was partially blocked by butoxycarbonyl-Phe-Leu-Phe-Leu-Phe (Boc2), a specific antagonist of the LXA(4) receptor (ALXR). Total 5-LOX expression did not change, regardless of treatment, but LXA(4) could inhibit nuclear translocation induced by MCAO or OGD. We also found that LXA(4) inhibits the upregulation of both leukotriene B(4) (LTB(4)) and leukotriene C(4) (LTC(4)) and the phosphorylation of extracellular signal-regulated kinase (ERK) induced by MCAO or OGD. The phosphorylation of the 38-kDa protein kinase (p38) and c-Jun N-terminal kinase (JNK) was not altered throughout the experiment. These results suggest that the neuroprotective effects of LXA(4) are probably achieved by anti-inflammatory mechanisms that are partly mediated by ALXR and through an ERK signal transduction pathway.

[Effect of lipoxin A(4) on lipopolysaccharide-induced endothelial hyperpermeability in human umbilical vein endothelial cell].

OBJECTIVE: To explore whether lipoxin A(4) (LXA(4))could prevent lipopolysaccharide (LPS)-induced human umbilical vein endothelial cells (HUVEC) monolayer hyperpermeability and its possible mechanism. METHODS: Human umbilical cords were obtained from women with normal pregnancy immediately after delivery from Tongji Hospital Affiliated of Tongji Medical College. Primary HUVEC were isolated from umbilical veins and subcultured, then, HUVEC were divided into four groups:control group; LPS group (10 mg/L of LPS); LPS + LXA(4) group(10 mg/L of LPS and 100 nmol/L of LXA(4)); LPS + LXA(4) + BOC-2 group [10 µmol/L of BOC-2, an effective antagonist of formyl peptide receptor like 1 (FPRL-1)]. All expriments were performed after cells were treated for 24 hours. Endothelial permeability was measured by fluorescein isothiocyan-ate labelled bovine serum albumin (FITC-BSA) clearance across the monolayer; tumor necrosis factor α (TNF-α) mRNA and secretion were detected by reverse transcriplase (RT)-PCR and ELISA assay respectively, and nuclear factor κB (NF-κB) protein change was determined by western blot. RESULTS: (1) LPS induced a significant increase in the permeability [Pa value of LPS group was (183.1 ± 1.7)%], while co-administrating with LXA(4) obviously attenuated this LPS-induced hyperpermeability, Pa value of LPS + LXA(4) group was (103.1 ± 2.2)%, LPS + LXA(4) + BOC-2 group was (162.2 ± 2.8)%, control group was 100%, the permeability of HUVEC monolayer was significantly increased by LPS which was (83.1 ± 1.7)% of control (P < 0.01), however, it was notably inhibited by LXA(4) (P < 0.05); the blockade of FPRL-1 could attenuate the effect of LXA(4), that is, there was no difference between the LPS + LXA(4) + BOC-2 group and the LPS group. (2) After treatment with different concentration of LPS(0, 0.1, 1, 10 mg/L), the mRNA expressions of TNF-α were increased (1.11 ± 0.11, 1.27 ± 0.03, 1.60 ± 0.06, 1.82 ± 0.04, respectively), compared with the control group, at the concentration of 1, 10 mg/L LPS, the difference was statistically significant (P < 0.05). (3) The increased levels of NF-κB and inflammatory mediator TNF-α in the LPS group were both inhibited by LXA(4). Levels of NF-κB protein and TNF-α mRNA secretion in LPS treated group (0.53 ± 0.06 and 0.81 ± 0.09, respectively) were both inhibited by LXA(4)(0.19 ± 0.05 and 0.41 ± 0.07, respectively, and both had significant difference, P < 0.05). (4) Levels of TNF-α in HUVEC culture medium of LPS group [(31.94 ± 0.01) ng/L] was significantly higher than the control group [(18.17 ± 0.03) ng/L, P < 0.05], LPS + LXA(4) group [(15.72 ± 0.07) ng/L] was significantly lower than the LPS group (P < 0.05). CONCLUSION: Our findings demonstrated that LXA(4) could prevent the endothelial cell hyperpermeability induced by LPS in HUVEC under which the possible mechanism was through inhibiting the expression of NF-κB and its related cytokines through receptor-dependent.

Evaluation of prognostic factors for synovial sarcoma / 中华外科杂志

<p><b>OBJECTIVE</b>To determine the independent prognostic factors of primary synovial sarcoma.</p><p><b>METHODS</b>The clinical data of 52 patients followed up from 66 patients with synovial sarcoma treated between September 1997 and September 2008 was analyzed retrospectively. There were 28 male and 24 female patients aged from 11 to 71 years old. Three and five-year overall survival (OS), recurrence rate and 9 prognostic factors were analyzed in this study. Univariate and multivariate analysis were performed to determine the prognostic factors of OS.</p><p><b>RESULTS</b>Fifty-two patients were followed up with the follow-up time ranged from 6 to 88 months (median 32 months). The 3-, 5-year overall survival rate and local recurrence rate were 52.8%, 30.3% and 32.7% respectively. Univariate showed tumor size < 5 cm, tumor located at extremities, adequate surgical margin and radical resection combined with radiotherapy had better survival rate (P < 0.05). Multivariate analysis demonstrated that tumor size, primary site and adequate surgical margin were independent prognostic factors for OS. Patients received radical resection combined with radiotherapy have longer median relapse time (25 months) compared with marginal resection combined with radiotherapy (18 months) and single radical resection (12 months). Thirty-five (67%) patients were treated with chemotherapy and seventeen (33%) patients received no chemotherapy for the primary tumor. Treatment with chemotherapy was not associated with an improved OS (P = 0.52).</p><p><b>CONCLUSIONS</b>The independent prognostic factors of synovial sarcoma are tumor size, primary site and adequate surgical margin. Doxorubicin and ifosfamide based chemotherapy was not associated with an improved OS in patients with synovial sarcoma. Radical resection combined with radiotherapy can best control local condition.</p>

[Effect of lipoxin A(4) on lipopolysaccharide-induced oxidant stress in human umbilical vein endothelial cells].

OBJECTIVE: To explore the effects of lipoxin A(4) (LXA(4)) on lipopolysaccharide (LPS)-induced oxidative stress in human umbilical veins endothelial cells (HUVEC) and the possible mechanism. METHODS: Neonatal umbilical cords were obtained from normal term pregnant women with cesarean section within 4 hours and then were used to isolate HUVEC for subculture. HUVEC were divided into four groups:control group; LPS group (10 µg/ml of LPS); LPS + LXA(4) group (10 µg/ml of LPS and 100 nmol/L of LXA(4)); LXA(4) group (100 nmol/L of LXA(4)). All expriments were performed after cells treated for 12 and 24 hours respectively. Immunofluorescence was used to detect the expression of VIII foctor and nuclear translocation of nuclear factor-erythroid-2-related factor 2 (Nrf2); the mRNA expression of Nrf2, heme oxygenase 1 (HO-1) and reduced form of nicotinamide-adenine dinucleotide quinone oxidoreductase-1 (NQO1) were evaluated by reverse transcription-PCR. RESULTS: (1) The flavovirens fluorescence was observed in the cytoplasm under fluorescence microscope, which confirmed the existence of VIII factor which specifically expressed in endothelial cells, especially in HUVEC. (2) Immunofluorescent results showed that in control group, Nrf2 protein expressed in the cytosol rather than in the nucleus. In LPS group, the expression of Nrf2 protein obviously increased in the nucleus while decreased in the cytosol after 12 hours. However, after LPS treatment for 24 hours, Nrf2 expression reduced in the cytosol and nucleus. In co-treatment with LPS and LXA(4) group, the expression of Nrf2 protein was much higher than that in LPS group after 12 hours or 24 hours. Furthermore, Nrf2 protein also mostly expressed in the cytosol in LXA(4) group. (3) After stimulation for 12 hours, compared with control group, the gene expression of Nrf2 and HO-1 were significantly enhanced in LPS group (0.581 ± 0.019 and 0.081 ± 0.009, P < 0.05) and in LPS + LXA(4) group (0.692 ± 0.048 and 0.136 ± 0.018, P < 0.05), the level of NQO1 mRNA in LPS group and LPS + LXA(4) group were 0.381 ± 0.009 (P > 0.05) and 0.574 ± 0.034 (P < 0.05). After treatment for 24 hours, compared with control goup, the gene expressions of Nrf2 and NQO1 were down-regulated in LPS group (0.180 ± 0.017 and 0.472 ± 0.064, P < 0.05). But in LPS + LXA(4) group the expression of Nrf2 and NQO1 were upregulated (0.532 ± 0.051 and 0.830 ± 0.068, P < 0.05, compared with treatment for LPS group). The mRNA expressions of Nrf2, HO-1 and NQO1 were increased in LPS + LXA(4) group compared with LPS group (P < 0.05). In addition, there was no markedly difference in the expressions of Nrf2, HO-1 and NQO1 between control and LXA(4) group after 12 hours and 24 hours (P > 0.05). CONCLUSION: Through activating nuclear translocation of Nrf2 protein from cytoplasm, LXA(4) upregulates the Nrf2 downstream enzymes, such as NQO1 and HO-1 to protect HUVEC against the oxidative stress induced by LPS.

Effects of exogenous annexin-1 on lipopolysaccharide-induced proliferation and reactive oxygen species production partially through modulation of CRAC channels but independent of NF-kappaB pathway.

OBJECTIVE: To investigate the effects of exogenous annexin-1 (ANXA1) on lipopolysaccharide(LPS)-induced proliferation, reactive oxygen species (ROS) production, and calcium signal transduction in RAW264.7 macrophages. METHODS: RAW264.7 macrophages were treated with or without LPS in the absence or presence of ANXA1. The proliferation effects were detected by Cell Counting Kit-8 assay. ROS were quantified by flow cytometry and fluorescence microscopy. Intracellular Ca(2+) concentration ([Ca(2+)](i)) was analyzed by laser confocal scanning microscopy. IkappaBalpha degradation and NF-kappaB translocation were tested by Western blot. RESULTS: Exogenous ANXA1 inhibited LPS-induced proliferation and ROS production in a dose-dependent manner. LPS evoked [Ca(2+)](i) increase through CRAC channels, and ANXA1 suppressed LPS-induced [Ca(2+)](i) increase in a dose-dependent manner. The CRAC channels were associated with LPS-induced proliferation and ROS production. Exogenous ANXA1 had no effect on LPS-induced IkappaB degradation and NF-kappaB translocation. CONCLUSIONS: ANXA1 inhibited LPS-induced proliferation and ROS production in RAW264.7 macrophages partially through modulation of CRAC channels but independent of the NF-kappaB pathway.

[Protective effects and its mechanism of lipoxins on human umbilical vein endothelial cells under hypoxia].

OBJECTIVE: To investigate protective effects and mechanisms of lipoxinA(4) (LXA(4)) on human umbilical vein endothelial cells (HUVEC) under hypoxia in vitro. METHODS: The HUVEC culture were divided into groups as followed: added M199 culture medium as normal control groups, added CoCl2 to mimic hypoxia in vitro as hypoxia group and added different concentrations of LXA(4) (1, 10, 100 nmol/L) were added to the induced hypoxial HUVEC as agents intervention group. Morphological changes of HUVEC were observed by using inverted phase contrast microscope. The influence of LXA(4) on cell survival was investigated by methyl thiazolyl tetrazolium (MTT) assaying method after the treatment with different concentrations of LXA(4) and 100 nmol/L lipoxinA(4) according to different time (4, 8, 12 and 24 hours). The expression of von-willebrand factor (vWF) was detected by immunocytochemistry method. The changes of cytosolic Ca(2+) were measured by laser scanning confocal microscope. RESULTS: (1) Morphological changes:the cells under hypoxia lost its normal shapes and showed necrosis, while the cells cocultured with 100 nmol/L LXA(4) were normal appropriately. (2) Survival rate: the survival rates of HUVEC under hypoxia was (40.1 +/- 3.9)% and increased to (52.9 +/- 1.4)%, (64.1 +/- 3.3)%, (76.6 +/- 1.6)% respectively when added with LXA(4) with concentration of 1, 10, 100 nmol/L into culture medium. There was significant different survival rate when compared with that of hypoxia group. (3) The level of vWF: The expression of vWF was decreased with the increasing concentrations of LXA(4) added into culture medium, the gray values were 203.9 +/- 0.7 in 1 nmol/L, 204.6 +/- 0.9 in 10 nmol/L, 191.8 +/- 0.5 in 100 nmol/L respectively, which reached statistical difference in comparison with that of hypoxia groups (P < 0.05). (4) Confocal analysis: the intracellular free Ca(2+) concentrations of HUVEC were intensified with LXA(4) treatment. CONCLUSIONS: LXA(4) plays an important role in keeping the normal shape of HUVEC under hypoxia, can enhance survival of hypoxial HUVEC and decrease the level of vWF in cytoplasm. The protective mechanism might be via decreasing mitochondria Ca(2+) overload and increasing cytoplasm Ca(2+) by nucleus Ca(2+) transference.

Lipoxin A(4) inhibited hepatocyte growth factor-induced invasion of human hepatoma cells.

AIM: Inflammation is a critical component of tumor progression. Lipoxin A(4) (LXA(4)) has been approved for potent anti-inflammatory properties. Recently, it was reported that LXA(4) repressed the expression and activity of cyclooxygenase-2 (COX-2), which is essential for invasion. However, there are few reports dealing with its effects on cancer. To explore whether LXA(4) regulate invasion, the effects of LXA(4) and its receptor agonist BML-111 on hepatocyte growth factor (HGF)-induced invasion of hepatoma cells and the possible mechanisms were researched. METHODS: Lipoxin A(4) receptor (ALX) expression in HepG2 cells were measured through reverse transcription polymerase chain reaction and western blot. Cytotoxicity of LXA(4) and BML-111 to HepG2 cells was detected by MTT and ((3)H)-TdR incorporation assay. Cell migration and invasion assays were performed using a Boyden chemotaxis chamber. COX-2 expression was detected by real-time polymerase chain reaction and western blot, respectively. Moreover, the expressions of matrix metalloproteinases (MMP)-2, MMP-9, IkappaBalpha and nuclear factor-kappaB (NF-kappaB) p65 were observed via western blot, and NF-kappaB transcriptional activity was tested by transfections and luciferase activities assay. RESULTS: ALX expression was detected in HepG2 cells, and suitable concentrations of LXA(4) and BML-111 had no cytotoxicity to cells. LXA(4) and BML-111 inhibited HGF-induced migration and invasion; downregulated COX-2, MMP-2 and -9; restrained HGF-induced IkappaBalpha degradation, NF-kappaB translocation and the transcriptional activity of NF-kappaB in HepG2 cells. Furthermore, exogenous PGE2 could reverse the inhibitory effects of LXA(4) also BML-111 on HGF-induced invasion and migration partially. CONCLUSION: LXA(4) inhibited HGF-induced invasion of HepG2 cells through NF-kappaB/COX-2 signaling pathway partially.