Background Arsenic is a human carcinogen. Arsenic and its metabolites affect the expression of p53, but whether there are any changes of p53 phosphorylation and ubiquitination levels in human bronchial epithelium cells (BEAS-2B) are not clear after exposure to arsenic and its metabolites. Objective To study the effects of arsenic and its metabolites monomethylarsic acid (MMA) and dimethylarsinic acid (DMA) on the expression of tumor suppressor gene p53 in BEAS-2B cells. Methods Different concentrations of sodium arsenite (NaAsO₂) were used to infect BEAS-2B cells, and the cell viability was detected with CCK-8 reagent to determine the dose and time of NaAsO₂ used for the following study. Based on the results of cell viability, the cells were divided into two panels: a sodium arsenide panel and an arsenic methylation metabolite penal. The doses of sodium arsenite were 0, 2, 4, and 6 μmol·L⁻¹ NaAsO₂; the arsenic methylation metabolite panel consisted of 0 μmol·L⁻¹ NaAsO₂ group (control), 6 μmol· L⁻¹ MMA group, 6 μmol· L⁻¹ DMA group， and 6 μmol· L⁻¹ NaAsO₂ group. The cells were collected after 48 h treatment, and the total protein and total RNA were extracted. The relative levels of p53 mRNA expression were determined by quantitative real-time polymerase chain reaction (qRT-PCR), the relative expression levels of p53 protein, p53 Ser9 and Ser15 phosphorylated proteins were determined by Western blot, and the level of p53 ubiquitination was detected by co-immunoprecipitation (CO-IP). Results Compared with the control group, the cell viability rates in all BEAS-2B cells treated by NaAsO₂ were significantly reduced (P<0.05), and the 50% cell viability was observed at 6 μmol·L⁻¹. Compared with the control group, the relative expression level of p53 mRNA gradually decreased after NaAsO₂ (2, 4, 6 μmol·L⁻¹) treatment (P<0.05), the relative expression levels of p53 protein and Ser9 phosphorylated protein induced by NaAsO₂ also decreased gradually (P<0.05), and the relative expression level of p53 Ser15 phosphorylated protein induced by NaAsO₂ followed the same pattern, but it was only lower than that of the control group in the 6 μmol·L⁻¹ NaAsO₂ group (P<0.05). Compared with the control group, there were no significant effects on the relative expression levels of p53 mRNA, p53 protein, Ser9 and Ser15 phosphorylated proteins in the MMA group and the DMA group. Compared with the control group, the expression level of p53 ubiquitination was significantly decreased and the expression of K48 ubiquitination decreased significantly after NaAsO₂ infection. Conclusion Arsenic causes a decrease in the expression of the p53 protein in BEAS-2B cells, largely due to inhibition of the phosphorylated pathway and a decrease in mRNA expression, and protein changes caused by a decrease in p53 ubiquitination do not play a dominant role. MMA and DMA do not affect p53 gene expression.

Objective To explore the value of serum IL-11 in the diagnosis and prognosis evaluation of acute cerebral infarction and its correlation with serum brain-derived neurotrophic factor(BDNF).Methods General clinical data of 102 patients with acute cerebral infarction(cerebral infarction group)and 64 normal controls(normal control group)were collected.According to the 90 d mRS score,the cerebral infarction group was divided into the good prognosis subgroup and the poor prognosis subgroup.Pearson correlation analysis was used to analyze the correlation between serum IL-11 and NIHSS score,cerebral infarction volume and serum BDNF.Logistics regression analysis was used to analyze the influencing factors of cerebral infarction prognosis,and the ROC curve of IL-11 in the diagnosis and prognosis of cerebral infarction was drawn.Results The rate of hypertension and the levels of glycosylated hemoglobin and low-density lipoprotein in the cerebral infarction group were significantly higher than those in the normal control group(all P＜0.05).The age,rate of diabetes,glycosylated hemoglobin level,NIHSS score at admission and cerebral infarction volume in the poor prognosis subgroup were significantly higher than those in the good prognosis subgroup(all P＜0.05).The level of serum IL-11 in the cerebral infarction group was significantly lower than that in the normal control group(t =10.123,P＜0.05).The serum IL-11 level in the poor prognosis subgroup of the cerebral infarction group was significantly lower than that in the good prognosis subgroup(t =7.438,P＜0.05).The expression of serum IL-11 in patients with cerebral infarction was negatively correlated with NIHSS score(r =-0.603,P＜0.001)and cerebral infarction volume(r =-0.681,P＜0.001).Logistics regression analysis showed that IL-11 was a protective factor(OR =0.814,P =0.009),while infarct volume(OR = 2.262,P＜0.001)and NIHSS score(OR =2.107,P =0.006)were risk factors affecting the prognosis of patients with cerebral infarction.When IL-11 was applied to the diagnosis of cerebral infarction,the area under the ROC curve was 0.841,the sensitivity was 91.18%,the specificity was 72.42%,and the cut-off value was 378.47;when IL-11 was applied to the prognostic discrimination of cerebral infarction,the area under the ROC curve was 0.786,the sensitivity was 67.09%,the specificity was 87.93%,and the cut-off value was 310.94.The correlation coefficient between serum IL-11 levels and serum BDNF levels in patients with cerebral infarction was r =0.711,P＜0.001.Conclusions The level of serum IL-11 in patients with cerebral infarction is significantly decreased,and the level of IL-11 in patients with poor prognosis is significantly lower than that in patients with good prognosis.Meanwhile,the level of IL-11 is negatively correlated with the level of serum BDNF,which may be used for the auxiliary diagnosis and prognosis evaluation of cerebral infarction.

OBJECTIVE To investigate the effects of catalpol on H2O2-induced osteoblast injury and its mechanism. METHODS The osteoblasts MC3T3-E1 were separated into control group， model group， empty group （transfected with empty plasmid）， catalpol group （100 μmol/L）， catalpol+forkhead box O3 （FoxO3） overexpression group （100 μmol/L catalpol+ transfected with FoxO3 overexpression plasmid）. After catalpol treatment and transfection， except for control group， other groups were induced with H2O2 to establish osteoblast oxidative stress model. The cell viability， apoptotic rate， alkaline phosphatase （ALP） activity， optical density （OD） value of calcium nodule， mean fluorescence intensity （MFI） of reactive oxygen species （ROS）， antioxidant enzyme activity [superoxide dismutase （SOD）， catalase （CAT）]， the levels of inflammatory factors [interleukin-6 （IL-6）， IL-1β]， and the expressions of FoxO3/Wnt/β-catenin signaling pathway-related proteins were detected in each group. RESULTS Compared with the control group， the cell viability， ALP activity， OD value of calcium nodule， activities of antioxidant enzyme， and the protein expressions of Wnt and β-catenin were decreased significantly in the model group， while apoptotic rate， MFI levels of ROS， inflammatory factor levels and the protein expression of FoxO3 were all increased significantly （P＜0.05）. Compared with the model group， above indicators of the empty group had no significant change （P＞0.05）， while those of catalpol group were reversed significantly （P＜0.05）. Compared with the catalpol group， the reversal effect of the changes in the above indicators was significantly weakened in the catalpol+FoxO3 overexpression group cells （P＜0.05）. CONCLUSIONS Catalpol can activate Wnt/β-catenin signaling pathway by down-regulating FoxO3， thereby inhibiting H2O2-induced MC3T3-E1 oxidative stress and inflammation reaction， enhancing cell viability and osteogenic differentiation activity， and alleviating apoptosis injury.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate factors associated with acute kidney injury(AKI) in postoperative colorectal cancer patients.Methods:The clinical data of 376 colorectal carcinoma (CRC) patients at Department of General Surgery, Affiliated Cancer Hospital of Zhengzhou University from Jan 2018 to Jun 2021 were retrospectively analyzed. Patients were divided into acute kidney injury (AKI) ( n=29) and non-AKI groups ( n=347). The demographic information, perioperative status, laboratory results and other relevant data of the two groups were compared . Binary logistic regression was used to analyze the independent risk factors for postoperative AKI. Results:Twenty-nine CRC patients (7.7%) had postoperative AKI. Binary Logistic regression analysis showed that preoperative hypertension ( OR=3.487, 95% CI: 1.081-11.251, P=0.037), anemia ( OR=3.158, 95% CI: 1.114-8.953, P=0.031), inadequate intraoperative crystalloid infusion ( OR=0.998, 95% CI: 0.997-0.999, P=0.007), low intraoperative mean arterial pressure ( OR=0.915, 95% CI: 0.863-0.970, P=0.003) and moderate to severe postoperative decline in hemoglobin levels ( OR=4.105, 95% CI: 1.487-11.335, P=0.006) were independent risk factors. Conclusion:Preoperative hypertension, anemia, inadequate intraoperative crystalloid infusion, low intraoperative mean arterial pressure, and moderate to severe postoperative decline in hemoglobin levels were independent risk factors for AKI development in colorectal cancer patients.

Ultrashort peptides have higher stability, tissue penetrability, biocompatibility, and less immunogenicity, and are widely applied in biology and medicine. GHK (glycyl-l-histidyl-l-lysine) and GQPR (glycyl-l-glutamyl-l-prolyl-l-arginine) can stimulate collagen renewal and inhibit collagen degradation. GHK and GQPR have been used in cosmetic anti-wrinkle skincare and make-up products. The most common approach for ultrashort peptide production is the solid-phase synthesis, which is eco-unfriendly due to heavy usage of organic chemical reagents during the manufacturing process. Here we report a new approach to the production of ultrashort peptides. Recombinant expression of ultrashort peptides is usually unfeasible because of the short amino acid sequences. A vector pET28a-Trxm harboring the thioredoxin gene was first constructed for subsequent fusion expression. The tandem repeats of GHK and GQPR genes were used as the templates for rolling circle amplification (RCA). The RCA reaction was tuned to incorporate noncanonical nucleotides 5-methylcytosine to obtain long DNA fragments. Gene sequences with various lengths were generated through double digestion of Acc65 Ⅰ and Apa Ⅰ. The resulting digestion products were gel recovered by size (from 500 bp to 1 500 bp) and cloned into pET28a-Trxm to obtain the recombinant vector pET28a-Trxm-(TRSP)_n. The pET28a-Trxm-(TRSP)_n was introduced into E. coli BL21(DE3) to generate a library of Trxm-(TRSP)_n sequences with a controlled distribution of lengths. Through double digestion and sequencing, positive clones with tandem repeats n=1, 2, 3, 4, 6, 7, 8, 9 were obtained. Protein expression results showed protein bands with corresponding molecular weight, and the protein expression level decreased as the tandem repeats increased. The expression level of Trxm-(TRSP)₁ achieved 50% of the total protein, while the expression level of Trxm-(TRSP)₂ was 30% of the total protein. The crude extracts from cell pellets were further treated with enterokinase cleavage, and the supernatants containing (TRSP)₁ were collected after ultrafiltration and then subjected to trypsin cleavage. HPLC analysis indicated that the ultrashort peptides GHK and GQPR were successfully obtained through two-step cleavage. This study may facilitate the commercial production of ultrashort peptides.
Escherichia coli/metabolism* ; Peptides/chemistry* ; Amino Acid Sequence ; Gene Library ; Tandem Repeat Sequences

Objective To study the clinical characteristics and analyze the correlative risk factors of interstitial pneumonia in systemic lupus erythematosus (SLE-IP).Methods 80 SLE patients in department of rheumatology of Nanfang hospital form January 2013 to January 2016 were retrospectively analyzed.SLE patients with interstitial pneumonia (n=40) were divided into case group.40 cases of SLE with interstitial pneumonia were selected and matched with age and sex.Patients with mild SLE without interstitial pneumonia were treated as controls.The clinical manifestations,routine examination,biochemical examination and immunological examination were performed to compare the risk factors of SLE-related interstitial pneumonia.Results In this study,non-specific interstitial pneumonia (NSIP) and usual interstitial pneumonia (UIP) were common in SLE-IP patients.the ground-glass opacities were more common in NSIP type,while Grid shadows and honeycomb shadows were more common in UIP type.The dry cough,chest tightness / shortness of breath,Raynaud's phenomenon,wet rales,triglyceride increased,anti-Sm antibody positive rate,anti-U1-nRNP positive rate between two groups were statistically significant (P＜0.05).Logistic regression analysis showed that the risk factors of SLE-IP were dry cough,chest tightness / shortness of breath,Raynaud's phenomenon,wet rales,triglyceride increased,anti-Sm antibody positive and anti-U1-nRNP positive.Conclusion The presence of dry cough,chest tightness / shortness of breath,Raynaud's phenomenon,wet rales,triglyceride increased,anti-Sm antibody positive and anti-U1-nRNP positive all suggest the probability of interstitial pneumonia in SLE patients.HRCT plays an important role in the diagnosis of interstitial pneumonia in lupus,which is valuable to improve the prognosis.

Objective The primary culture of synovial fibroblasts is a convenient tool to study the pathology and physiology of synovial tissues .An improved method was constructed in this study by C57BL /6 mice to study the mechanism of rheumatoid ar-thritis(RA) .Methods The synovium around the hip joints were collected .Attention should be paid to eliminate the egg-yolk like yellow oval substance in the middle of the synovium .The synovium was transferred into a 1 .5 mL Eppendorf tube containing 0 .5%type Ⅳ collagenase and cut into 1 mm3 blocks or so .The Eppendorf tube was placed in 37 ℃ Constant temperature orbital shaker incubator for 60 min .After digestion ,the tube was placed on the Vortex for a high-speed oscillation for 1 .5 minutes to guarantee the separation of cells .Results Within about 1 week ,the first passage was performed by the trypsin digestion method .On day 10 , the number of synovial macrophages reached the maximum and then decreased gradually .After the third generation (day 15 to 20) , the synovial macrophages generally disappeared .Vimentin was suitable for the immunofluorescence cytochemical staining for the synovial fibroblasts .The cell purity was indicated as > 95% .The cytometric analysis indicated that purity of Vimentin and CD90 .2-labelled cells was over 95% ;the purity of CD54-labelled cells was 80% approximately .Conclusion It is a simple and effective method for primary culture of synovial fibroblasts in mice .

Objective To learn the effect of breast removal of polyacrylamide hydrogel by areola incision;and to provide an effective method for removal of polyacrylamide hydrogel.Methods According to the accepted operation,45 cases with breast polyacrylamide hydrogel were divided into 25 cases of observation group and 20 cases of control group.The observation group was treated with areola medial semicircular incision breast removal of polyac-rylamide hydrogel,and the control group was treated with areola small incision suction to remove polyacrylamide hydrogel.The operative time,intraoperative blood loss,postoperative drainage tube extraction time,postoperative 7d breast shape and perioperative complications of the two groups were observed;postoperative follow -up was performed to evaluate the effect of surgery.Results The operation time of the observation group (190.2 ±36.1)min was longer than (119.5 ±44.0)min of the control group,the difference was statistically significant between the two groups (t =5.924,P <0.05);the intraoperative blood loss of the observation group (250.4 ±23.9)mL was higher than (89.7 ± 30.4)mL of the control group,the difference was statistically significant (t =19.865,P <0.05 ).The difference between postoperative 7d breast normal morphology rates of the two groups was not statistically significant (χ2 =2.571,P >0.05);the postoperative drainage tube extraction time of the observation group (3.7 ±0.5)d was more than (2.3 ±0.3)d of the control group,the difference was statistically significant (t =11.021,P <0.05 ).The complication rate of the observation group was 12.0%(3 /25),which of the control group was 10.0%(2 /20),the difference was not statistically significant (χ2 =0.045,P >0.05).All cases of the two groups were followed up;the mean follow -up time was (6.7 ±2.1)months.The good rate of surgery of the observation group was 92.0 % (23 /25),which of the control group was 70.0%(14 /20),the observation group was higher than the control group (χ2 =5.718,P <0.05).Conclusion For patients with complications after polyacrylamide hydrogel injection for augmentation mammoplasty;the breast polyacrylamide hydrogel removal surgery by areola medial semicircular incision is recommen-ded as the preferred.

BACKGROUND:Fisher-Lewis rat kidney transplant models are the international common chronic renal alograft rejection models, but their application is greatly limited because of difficulty in model preparation and high costs. OBJECTIVE:To explore a new method of establishing SD-Wistar rat models of chronic renal alograft rejection. METHODS: Fifty-six pairs of SD-Wistar rats were subjected to left kidney orthotopic transplantation. The right kidneys of the recipients were intact and used as internal controls. 23 rat recipients were randomly divided into model group (n=15) and control group (n=8). The rats in the model group were injected with cyclosporine microemulsion for 10 days (2 mg/kg/day,i.p.) after kidney transplantation. The rats in the control group were not treated with immunosuppressive therapy. RESULTS AND CONCLUSION:The irreversible acute rejection occurred in al the transplanted kidneys of rats in the control group within 4 weeks, leading to the necrosis of transplanted kidney. Moderate inflammatory cel infiltration appeared in the transplanted kidneys of rats in the model group at 4, 8 and 12 weeks after transplantation. Typical histopathological changes of chronic rejection were observed within 12 weeks after transplantation. The Banff total scores were increased with time after transplantation. Al these histopathological changes were not observed in the intact right kidneys of rat recipients in both groups. The valey value of