This study aims to evaluate the accuracy of portable hemoglobinometer (Hemocue Hb 201+ hemoglobin analyzer) in patients with maintenance hemodialysis (MHD) and its diagnostic value for anemia. The data of venous hemoglobulin (Hb) and fingertip capillary hemoglobulin (DHb) in MHD patients from Lingnan Hospital, the Third Affiliated Hospital of Sun Yat-sen University were retrospectively analyzed, and the correlation and difference between DHb and Hb and the accuracy of DHb in the diagnosis of anemia were evaluated. A total of 105 patients were included in the study. There was no significant difference between the paired DHb and Hb [(109±21) g/L vs. (108±20) g/L, t=-1.284, P=0.202]. Pearson correlation analysis showed that DHb was positively correlated with Hb ( r=0.929, P<0.001). Linear regression analysis showed that DHb and Hb met the regression equation Hb=0.88×DHb+12.23, and P<0.001. Bland-Altman analysis showed that the differences between the paired DHb and Hb was (1.0±7.8) g/L with the limit of agreement as (-14.2, 16.2) g/L. The mean percentage of the differences in Hb was 1% with limit of agreement as (-13.7%, 15.7%). A DHb of >110 g/L was 0.90 sensitive and 0.83 specific to identify patients with an Hb >110 g/L and its positive and negative predictive values were 0.84 and 0.90, respectively. It suggests that, in MHD patients, Hemocue Hb 201+ analyzer shows good accuracy, and can be used to monitor the Hb trend and serve as a screen method for those reaching target Hb.

AIM: To detect the expression of P53 and mTOR in pterygium tissues and healthy conjunctival tissues, and to explore the relationship between the expression of P53 and mTOR, and the relationship between the expression of P53 and mTOR and the important clinical features of pterygium.METHODS: The surgical specimens of 43 patients(43 eyes)who underwent pterygium excision and autologous conjunctival transplantation in the First Affiliated Hospital of Shihezi University from November 2022 to May 2023 were collected. Healthy conjunctiva group was selected from the healthy conjunctival tissue that originated from the temporal conjunctiva of 13 patients. Totally 10 pterygium specimens and 6 normal conjunctival specimens were selected and the qPCR was used to detect the mRNA expression levels of P53 and mTOR in pterygium and normal conjunctival tissues. Another 33 cases of pterygium and 7 cases of normal conjunctival tissues were collected and the expression of P53 and mTOR in pterygium and normal conjunctival tissues were detected by immunohistochemistry. IPP6.0 software was used to calculate the average optical density, the correlation between the expression levels of P53 and mTOR, and the correlation between the expression levels of P53 and mTOR and the important clinical features of pterygium were analyzed.RESULTS: According to qPCR results, the mRNA expression levels of TP53 and mTOR in the pterygium group were significantly higher than those in the healthy conjunctiva group(all P<0.05). According to the immunohistochemical staining results, the expression levels of P53 and mTOR proteins in the pterygium group were significantly higher than those in the healthy conjunctiva group(P<0.05). The expression of P53 was positively correlated with the expression of mTOR(r=0.417, P<0.05). The expression of P53 in the group of outdoor activity time > 3 h was higher than that in the group of outdoor activity time ≤3 h(P<0.05). The expression of P53 in the group of pterygium head invasive limbal distance > 2 mm was higher than that in the group of pterygium head invasive limbal distance ≤2 mm(P<0.05). There was no difference in the expression of pterygium between the two groups of patients aged > 40 years and ≤40 years(P>0.05). There was no significant difference in the expression of mTOR between the groups of outdoor activity time > 3 h and ≤3 h, the group of pterygium head invasion distance > 2 mm and ≤2 mm, and the group of > 40 years old and ≤40 years old(all P>0.05). The expression of P53 was positively correlated with the duration of outdoor activities(r=0.484, P<0.01)and the distance of limbal invasion(r=0.479, P<0.01). The expression of mTOR was not correlated with age, duration of outdoor activities, and distance of limbus invasion(all P>0.05).CONCLUSION: The overexpression of P53 and mTOR in pterygium showed a positive correlation, suggesting that the abnormal expression of P53 and mTOR may play a role in the pathogenesis of pterygium, which provides an experimental basis for further exploring the pathogenesis of pterygium; the expression of P53 is positively correlated with the time of outdoor activities and the distance of pterygium invasion. The P53 plays a role in evaluating the severity of pterygium, and provides new ideas for the clinical diagnosis and treatment of pterygium.

ObjectiveTo investigate the mechanism of Ershiwuwei Guijiu pill in preventing and treating postmenopausal osteoporosis （PMOP） by activating the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway and inhibiting excessive autophagy. MethodFemale SD rats were ovariectomized and randomly divided into the sham operation group （Sham）， the operation group （OVX）， the Ershiwuwei Guijiu pill （GJ） group， and the raloxifene hydrochloride （RLX） group， with 10 rats in each group. Enzyme-linked immunosorbent assay （ELISA） and colorimetric methods were used to detect the levels of estrogen， bone metabolism markers in serum， and total superoxide dismutase （T-SOD）， malondialdehyde （MDA）， and glutathione peroxidase （GSH-Px） in tibial tissue. Flow cytometry was used to detect reactive oxygen species （ROS） levels in bone marrow mesenchymal stem cells. Masson staining was used to observe pathological changes in the proximal tibia， and micro-computed tomography （Micro-CT） was used to observe changes in tibial microstructural parameters. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the expression of autophagy-related proteins Beclin1， microtubule-associated protein 1A/1B-light chain 3 （LC3）， autophagy-related 5 （Atg5）， as well as PI3K， Akt， and mTOR in tibial tissue. ResultCompared with the Sham group， the OVX group showed a significant decrease in serum levels of estradiol （E₂） and calcium ion （Ca²⁺）， and T-SOD， GSH-Px， PI3K， Akt， and mTOR mRNA levels in bone tissue （P<0.05， P<0.01）， significantly reduced bone mineral density （BMD）， bone surface/bone volume （BS/BV）， trabecular thickness （Tb.Th）， trabecular number （Tb.N）， and trabecular connectivity （Con） in the tibia （P<0.05， P<0.01）， thinner epiphyseal growth plate， and the bone marrow cavity filled with fat vacuoles. Moreover， the levels of phosphorus （P）， MDA， ROS， and mRNA and protein expression of Beclin1， LC3， and Atg5， as well as trabecular separation （Tb.Sp） were significantly elevated （P<0.05， P<0.01）. Compared with the OVX group， the GJ and RLX groups showed significant increases in serum E₂ and Ca²⁺， and bone tissue levels of SOD， GSH-Px， and the mRNA levels of PI3K， Akt， and mTOR （P<0.05， P<0.01）， significantly increased BMD， BS/BV， Tb.Th， Tb.N， and Con in the tibia， thickened epiphyseal growth plate， and significantly reduced fat vacuoles in the bone marrow cavity （P<0.05， P<0.01）. Additionally， the levels of P， MDA， ROS， Beclin1， LC3， Atg5 mRNA and proteins， and Tb.Sp were significantly decreased （P<0.05， P<0.01）. The ratios of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR， which were significantly reduced in the OVX group （P<0.01）， were significantly increased in the GJ and RLX groups （P<0.01）. ConclusionThe Ershiwuwei Guijiu pill reduces oxidative stress and inhibits autophagy， thereby preventing and treating postmenopausal osteoporosis. Its mechanism may be related to the activation of the PI3K/Akt/mTOR signaling pathway， which inhibits autophagy.

Objective:To investigate the effect of oroxylin A (OrA) on macrophage pyroptosis induced by Mycobacterium tuberculosis ( Mtb) infection and the underlying molecular mechanism. Methods:An in vitro infection model was constructed by infecting J774A.1 cells with Mtb. The MTT method was used to detect the effect of OrA on the viability of J774A.1 cells. Lactate dehydrogenase (LDH) assay kit was used to detect the release of LDH by J774A.1 cells. Western blot was used to detect the protein levels of pyroptosis-related proteins such as NOD-like receptor thermal protein domain associated protein 3 (NLRP3), GSDMD-N, caspase1-p20, apoptosis-associated speck-like protein containing a CARD (ASC), and α7 nicotinic acetylcholine receptor (α7nAChR) as well as the phosphorylation of NF-κB p65. ELISA was used to detect the levels of IL-1β in each group. Immunofluorescence was performed to detect the expression and localization of α7nAChR. Results:MTT results showed that OrA had no cytotoxicity to J774A.1 cells at concentrations below 80 μmol/L. OrA reduced the release of IL-1β, down-regulated the expression of NLRP3, GSDMD-N and caspase1-p20 at protein level, inhibited ASC oligomerization, promoted the expression of α7nAChR at protein level, and inhibited the phosphorylation of NF-κB p65. In the presence of α7nAChR agonist PNU282987, the protein levels of GSDMD-N decreased and the phosphorylation of NF-κB p65 was inhibited.Conclusions:OrA may inhibits Mtb infection-induced macrophage pyroptosis through regulating cholinergic anti-inflammatory pathway.

Objective:To construct the expression vector of the fusion protein of human serum albumin (HSA) and thymopentin (TP5) and to express it in Pichia pastoris,and to elucidate the biological activity of fusion protein.Methods:The HSA-TP5 fusion gene was constructed by gene recombination and transfected into Pichia pastoris to construct the eukaryotic expression system of HSA-TP5.The recombinant eukaryotic expression plasmid of PPICZα-HSA-TP5 was obtained by agarose gel electrophoresis and purification reagent.The two step fermentation method was used to ferment gene engineering bacteria of HSA-TP5 in high density,and the fermentation supernatant protein was precipitated and concentrated;the purified fusion protein was obtained by cation exchange chromatography and hydrophobic chromatography and analyzed by SDS-polyacrylamide gel electrophoresis.The effect of the fusion protein on the proliferation of lymphocytes was detected by MTT assay.Results:The HSA target gene fragment with length of 1 845 bp was achieved by PCR method.The HSA-TP5-pPICZαC fusion plasmid was identified by restriction endonuclease digestion,and the fragment length was 707 bp.The sequence analysis showed that the HSA and TP5 sequences of the target genes were identical with the gene sequences reported in GenBank and were fused by forward fusion.PCR method confirmed that the eukaryotic recombinant plasmid PPICZ αC-HSA-TP5 was integrated into the yeast genome,and compared with control group,the target gene PCR product length was found to be 1 860 bp.SDS-PAGE analysis showed that the expression level of HSA-TP5 fusion protein was gradually increased with the induction time within 72 h.HSA-TP5 fusion protein was purified by cation exchange chromatography and AKTA multifunctional protein purification system.The MTT assay results showed that HSA-TP5 fusion protein was consistent with TP5 protein in promoting lymphocyte proliferation activity.Conclusion:HSA-TP5 fusion protein can be obtained by constructing the eukaryotic expression system of Pichia pastoris and owns the biological activity.

Objective To validate the value of Cleveland Clinical Score in predicting acute renal injury requiring renal replacement therapy(RRT-AKI) after cardiac valve surgery in Chinese adult patients.Methods An analysis was conducted for all the adult patients who underwent cardiac valve surgery from January 2010 to December 2014 in Changhai Hospital,Shanghai.A total of 3 230 adult patients were included.Based on Cleveland Clinical Score,the patients were divided into 3 risk stages:0 to 2 point,3 to 5 point,and 6 to 8 point.The incidence of RRT-AKI were compared between different stages.And the predictive value of the Cleveland Clinical Score model was assessed by area under the receiver operating characteristic curve(AUC-ROC) and the model calibration was assessed using the Hosmer-Lemeshow test.The patients were also divided into two groups:Non-RRT group and RRT-AKI group.The mortality were compared between these two groups.Results The incidence of RRT-AKI was 1.67％ vs the predicted ratio of RRT-AKI 1.70％ (x2 =0.018,P =0.892).Among the stage 1,2,and 3,the actual incidence of RRT-AKI,was 1.23％,2.66％,and 16.7％ vs the predicted incidence 0.40％,1.80％,and 9.50％,respectively.The AUC-ROC for Cleveland Clinical Score predicting RRT-AKI was 0.64 [95 ％ CI(0.57,0.71),P ＜0.01].Compared with Non-RRT group,the RRT-AKI group got a higher mortality(87.00％ vs 1.50％,x2 =1 330,P ＜0.01).Conclusion The Cleveland Clinical score had no real predictive value for RRT-AKI in Chinese adult patients after cardiac valve surgery.The incidence of RRT-AKI of the whole population and the stage 3 patients could be predicted by the model.And the patients with a high Cleveland score got a higher mortality than that of patients with a low Cleveland score.

Objective To validate the value of Simplified Renal Index Score(SRI) in predicting acute renal injury requiring renal replacement therapy(RRT-AKI) after cardiac valve surgery in Chinese adult patients.Methods An analysis was conducted for all the adult patients who underwent cardiac valve surgery from January 2010 to December 2014 in Changhai Hospital,Shanghai.A total of 3 183 adult patients were included.Based on SRI Score,the patients were divided into 3 risk stages:0 to 1 point,2 to 3 point,and 4 to 8 point.The incidence of RRT-AKI was compared between different stages.And the prediction value of the SRI model was assessed by area under the receiver operating characteristic curve (AU-ROC) and the model calibration was assessed with the Hosmer-Lemeshow (H-L) test.Results After surgery 52 (1.6％) patients developed acute kidney impairment and subsequently underwent renal replacement therapy.Patients with low values of simplified renal index (0-1),medium(2-3) and high values (4 and more) were found to have increasingly higher risk for renal replacement therapy of 0.8％ (95％ CI:0.005-0.012) 、3.8％ (95％ CI:0.026-0.052) 、20％ (95％ CI:0.010-0.720),respectively.TheAU-ROCwas0.68(95％ CI:0.610-0.760,P＜0.01).The H-L test was x2 =2.45,P=0.29.Conclusion SRI model gives a certain clinical significance,suggesting that high-values patients may occur RRT-AKI with a significantly higher risk than low-values patients.However,SRI model cannot give an accurate prediction value for RRT-AKI in Chinese adult patients after cardiac valve surgery.Direct clinical use of the model should be considered cautiously.

An on-line solid phase extraction ( SPE ) coupled with HPLC-MS/MS method was developed to determine S-ammuxetine and R-ammuxetine in rat plasma. The sample preparation consisted of the following steps:A protein precipitation extraction used methanol and acetonitrile ( 50:50 , V/V ); an on-line SPE treatment to remove most matrixes in plasma;an enrichment and separation step used a C18 analytical column. S-and R-ammuxetine were determined by tandem mass spectrometry. The SPE column was a Retain PEP Javelin (10 mm × 2. 1 mm × 5 μm), while the chromatographic separation was achieved using a ZORBAX SB-C18 (50 mm × 2. 1 mm × 3. 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (40:60:0. 1, V/V/V, 0. 3 mL/min). The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 292 . 1/154 . 0 and m/z 260. 4/116. 2 were used to measure S-ammuxetine, R-ammuxetine and internal standard (propranolol). The method was linear over a concentration range from 0 . 2 to 1000 μg/L with the correlation coefficients of 0 . 9903 and 0 . 9951 . The average intra-day precision values were 1 . 2% -12 . 0% for S-ammuxetine and 0. 4%-11. 2% for R-ammuxetine, respectively. The average recoveries were 94. 2%-101. 6% for S-ammuxetine and 94. 3% -109. 4% for R-ammuxetine. Compared to the literature, the sensitivity of this method increased dramatically. The present method has been successfully applied to the preclinical rat research of ammuxetine isomers following intragastric administration.

BACKGROUND:Our preliminary study found that the monocusp valves made of ultramicropore expanded
polytetrafluoroethylene (ePTFE) revealed no significant thrombus, calcification, or degradation 20 weeks after implanted into the descending aorta and the left pulmonary artery in sheep, which verified the good property of ePTFE. However, the surface of ePTFE in the left pulmonary artery was covered with obvious neointima.
OBJECTIVE: To assess the biocompatibility of phosphorylcholine-coated ePTFE.
METHODS:ePTFE surface was modified by phosphorylcholine derivative. Then the changes of surface shape, tensile stress at yield and elasticity modulus, water contact angle, and protein absorption capacity of ePTFE after surface modification were observed. (1) Hemolytic test: the leaching solution of phosphorylcholine-coated ePTFE, leaching solution of uncoated ePTFE, normal saline, and distiled water were added to the diluted human blood, respectively. (2) Platelet count test: the phosphorylcholine-coated ePTFE, uncoated ePTFE, high density
polyethylene, and Zymosan A were added to the whole blood samples from healthy volunteers, respectively.
(3) Platelet activation test: the phosphorylcholine-coated ePTFE, uncoated ePTFE, γ-Globulins, and Zymosan A
were added to the whole blood samples from healthy volunteers, respectively.
RESULTS AND CONCLUSION: The mean micropore diameter of ePTFE was significantly decreased after
phosphorylcholine coating (P < 0.001). The hydrophilicity and the ability of suppressing protein adsorption were
significantly strengthened after phosphorylcholine coating (P < 0.001). Phosphorylcholine coating did not influence
ePTFE in biomechanical properties and hemolytic test. The platelet count test and platelet activation test demonstrated that phosphorylcholine coating significantly improved anti-thrombus function of ePTFE. So, phosphorylcholine coating can enhance anti-thrombus function, suppress protein adsorption, and improve biocompatibility of ePTFE.

Objective To establish an evaluation system about animals infected with hantavirus,an observation of the BALB/c mice infected with hantavirus was made.Methods BALB/c mice were infected with hantavirus by intramuscular injection with stock solution.The specific antigen from BALB/c mice tissues after 3 days was detected with enzyme-linked immunosorbent assay (ELISA) and viral RNA with real-time polymerase chain reaction (RT-PCR).Results Within a short term,the specific antigen and viral RNA were detected from the brain and liver at day 3 after infection,but not be detected from the heart,spleen,lung,and kidney samples.Conclusions The results provided ones with some information on animals infected with hantavirus.