[ ABSTRACT] AIM:To investigate the inhibitory effect of ginsenoside Re on intimal hyperplasia induced by bal-loon-injury and to explore the role of NF-κB p65 signaling pathway in the process.METHODS:SD rats (n=40) were di-vided into 5 groups randomly: sham operation group, model group, low-dose ginsenoside Re group, middle-dose ginsen-oside Re group and high-dose ginsenoside Re group.The carotid artery intima injury model was established by 2F balloon catheters in all groups except the sham operation group.The day after modeling, the animals in model group and sham op-eration group were administered intragastrically with distilled water, and the rats in low-dose, middle-dose and high-dose ginsenoside Re groups were given ginsenoside Re at doses of 12.5 mg/kg, 25mg/kg and 50 mg/kg, respectively.After 14 continuous days, the morphological changes of the injured arteries were observed by HE staining and the lumen area, intima area and media area as well as the ratio of intimal area/media area were determined.The expression of tumor necrosis fac-tor-α( TNF-α) and interleukin-1β( IL-1β) were detected by real-time PCR.The proliferating cell nuclear antigen ( PC-NA) and nuclear factor-kappa B ( NF-κB) p65 were examined by immunohistochemistry.RESULTS:Compared with sham operation group, the vessel cavity was narrowed (P<0.01), the mRNA levels of TNF-αand IL-1β, and the protein ex-pression of PCNA and NF-κB p65 were increased in model group (P<0.05).Compared with model group, the vascular intimal hyperplasia was alleviated obviously (P<0.05), and the mRNA levels of TNF-αand IL-1β, and protein expres-
sion of PCNA and NF-κB p65 were decreased in medium and high-dose ginsenoside Re groups (P<0.05).CONCLU-SION:Ginsenoside Re inhibits the vascular neointimal hyperplasia induced by balloon-injury in rats, and the molecular mechanism may be related to the inhibition of NF-κB p65 signaling pathway.

Objective To investigate the clinical significance of IL-27 in hepatocellular carcinoma ( HCC) in the early stage , and to compare it with alpha-fetoprotein ( AFP) in diagnostic performance .Methods Enzyme-linked immunosorbent assay was used to detect the serum level of IL-27 in the HCC group and control group .Receiver operator characteristic ( ROC) curve was used to ana-lyze two indicators and the logistic regression model predicted value ( PRE) was obtained .Results The study indicated the concentra-tion of IL-27 in HCC group [(364.19 ±177.55)pg/ml] was significantly higher than the control group [(255.49 ±94.33)pg/ml], the area under the curve (AUC) of IL-27 and AFP were (0.804) and (0.818), respectively.Logistic regression obtained the regres-sion model PRE that contains AFP and IL-27, with a predicted area under the ROC curve for HCC (0.901), while the diagnostic sen-sitivity (84.8%) and specificity (96.7%) were significantly higher than the capability of individual diagnosis of each indicator .Con-clusions In this study we obtained the model which can be used for clinical diagnosis of hepatocellular carcinoma in future .

Objective To investigate the effect of combined antiplatelet drugs on clinic and aggregation activity of platelet in patient with aucte myocardial infarction(AMI).Methods Consecutive 97 cases with AMI were redomized to receive clopidogrel plus aspirin(treated group,n=48) or aspirin alone(placebo group,n=49) for 30 days.Before and after treatment,the clinical recurrent angina,improvement of heart function,death,ST segment changes of electrocardiogram and the rate of maximun aggregation of paltelet were tested and analysed.Results Recurrent angina occurred in 10.0%(5 cases) in the treated group and 20%(10 cases) in the placebo group.The total frequency of improvement of heart function occurred in 36.7%(18 cases) in the treated group and 22.4%(11 cases) in the placebo group(?~2=3.638,P=0.045).Death occurred in 4%(2 cases) in the treated group and 8%(4 cases) in the placebo group.ST segment of electrocardiogram decreased from (0.36?0.13)mV to (0.13?0.08)mV,compared with placebo from (0.35?0.14)mV to (0.16?0.90)mV(t=3.012,P=0.04).The rate of maximum aggregation of platelet induced by ADP reduced from (74.54?8.99)% to (34.09?9.23)%,the placebo from (72.30?7.78)% to (56.54?6.92)%(t=13.42,P

BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.

OBJECTIVETo evaluate the impact of the biomarkers and the clinicopathological characteristics on the prognosis of malignant melanoma (MM).

METHODSThe clinical data of 127 MM cases were retrospective analyzed. The surgical specimens of MM were analyzed with immunohistochemistry for detecting HMB45, S-100 and vimentin expressions, and univariate and multivariate regression analysis was performed to analyze their correlation to the prognosis of the patients.

RESULTSAmong the 127 MM cases, the positivity rates of HMB45, S-100 and vimentin were 89.8%, 92.1% and 78.0%, respectively. Univariate analysis showed that the patients' age, ulcer, Clark classification, postoperative tumor margin, AJCC, treatment outcomes, and S-100 were significantly correlated to the prognosis, and multivariate analysis indicated that age, Clark classification, S-100, tumor margin and outcomes were the independent predictive factors for the prognosis of MM.

CONCLUSIONS-100, age, Clark classification, S-100, tumor margin and treatment outcomes were the independent prognostic factors for MM, and HMB45 and vimentin have no predictive value in the prognosis of MM.

Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Female ; Humans ; Male ; Melanoma ; diagnosis ; mortality ; pathology ; Melanoma-Specific Antigens ; metabolism ; Middle Aged ; Multivariate Analysis ; Prognosis ; Retrospective Studies ; S100 Proteins ; metabolism ; Skin Neoplasms ; diagnosis ; pathology ; Survival Rate ; Vimentin ; metabolism

To explore the role of dentritic epidermal T lymphocytes ( DETCs) in immune rejection of skin allograft in mice and its related mechanism. Methods (1) Full-thickness skin was harvested from back of one male wild type (WT) C57BL/6 mouse. Epithelial cells were isolated for detection of the expression of DETCs and their phenotype with flow cytometer. Another male WT C57BL/6 mouse was used to harvest full-thickness skin from the back. Epidermis was isolated for observation of the morphological characteristics of DETCs with immunofluorescence technology. (2) Four male green fluorescence protein (GFP)-marked C57BL/6 mice, 7 female WT C57BL/6 mice (group WT), and 7 female ybT lymphocytes 8 gene knock-out (GK) C57BL/6 mice (group GK) were used. Full-thickness skin in the size of 1.4 cm x 1.4 cm on the back of mice in groups WT and GK were excised, and the wounds were transplanted with full-thickness skin in the size of 1.2 cm x 1.2 cm obtained from male GFP-marked C57BL/6 mice. The survival time of skin grafts was affirmed with small animal in vivo imager and naked eyes and recorded. (3) Two male WT C57BL/6 mice were used to isolate epithelial cells. Cells were inoculated into 48-well plate and divided into activation group (A) and control group (C) according to the random number table, with 4 wells in each group. Cells in group A were treated with 10 pL concanavalin A in the concentration of 2 microg/mL for 24 hours, while those in group C with PBS in the same volume as that in group A. The expression of interferon y in DETCs was detected with flow cytometer. (4) Four male GFP-marked C57BL/6 mice were used as donors. Fourteen female WT C57BL/6 mice were used as receptors and divided into interferon gamma neutralizing group (IN) and control group (C) according to the random number table, with 7 mice in each group. The skin transplantation model of C57BL/6 male to C57BL/6 female was established as in part (2). Before surgery and 72 hours after, mice in group IN were intraperitoneally injected with 200 pL interferon y neutralizing antibody in the concentration of 1 mg/mL, and those in group C with normal saline in the same volume as that in group IN. The survival time of skin grafts was observed and recorded using the methods in part (2), and the result of group IN was compared with that of group GK in part (2). The survival curve of skin grafts was processed with Log-rank ( Mantel-Cox) test. Results (1) The positive expression rate of DETCs in epithelial cells of skin in mouse was 7.27%, and they were all CD3 cells. DETCs were found to be scattered in the epidermis of skin in mouse with dendritic morphology. (2) The survival time of skin grafts of mice in group GK was 22-35 d, obviously longer than that in group WT (12-16 d, y2 = 14. 10 , P < 0.001). (3) Expression of interferon gamma was detected in 22. 70% DETCs in group A, which was obviously higher than that in group C (0.51%). (4) The survival time of skin grafts of mice in group IN was 19-24 d, which was obviously longer than that in group C (12-16 d, chi 2 = 13.60, P < 0.001) but close to that in group GK as in part (2) (chi2 = 0.06, P = 0.810). Conclusions DETCs are involved in promotion of immune rejection of skin allograft probably by secretinf interferon gamma.
Allografts ; Animals ; Epidermis ; Female ; Graft Survival ; immunology ; physiology ; Interferon-gamma ; immunology ; metabolism ; Lymphocytes ; Male ; Mice ; Mice, Inbred C57BL ; Skin ; Skin Transplantation ; T-Lymphocytes ; immunology

BACKGROUNDTo evaluate the diagnostic values of multiple tumor marker protein biochip detective system for lung cancer.

METHODSThe serum levels of 12 tumor markers, including CA199, NSE, CEA, CA242, CA125, CA153, AFP, ferritin, free-PSA, PSA, β-HCG and HGH, were measured in 108 lung cancer patients, 48 patients with benign pulmonary lesion and 145 healthy by the detective system.

RESULTSThe positive rates were 83.33% (90/108), 52.08% (25/48) and 28.97% (42/145) in lung cancer, benign pulmonary lesion and healthy groups, respectively. The lung cancer group had significantly higher positive rate than that of the controls (Chi-Square=16.75 and 73.32, both P < 0.001); There was significant difference of positive rate in various clinical stages of lung cancer (Chi-Square=7.89, P=0.048), but not in different pathologic classification. Serum CA199, CEA and CA242 levels were closely correlated with clinical staging (F=2.84, P=0.041; F= 3.49, P=0.018; F =5.22, P=0.002). The positive rate of CEA in adenocarcinoma was higher, but no significant difference was observed (Chi-Square=0.71, P=0.07). NSE in small cell lung cancer had the highest positive rate (Chi-Square=19.03, P < 0.001). Combined measurement of the twelve markers had higher sensitivity (Chi-Square= 368.58, P < 0.001), but less specificity (Chi-Square= 369.87, P < 0.001).

CONCLUSIONSCombined measurement of various serum tumor markers using protein biochip can significantly increase the diagnostic sensitivity for lung cancer. Meanwhile, it is also significant for defining clinical stage, identificating pathologic classification, as well as monitoring therapeutic efficacy. As its specificity and positive predictive value are lower, it is more suitable to be used as a surveying tool for symptomless people, especially for high risk people for lung cancer.

Objective To study the biological effects of (Janus Kinase-Signal transducers and activators of transcription,AK-STATs)path-way in hair cell apoptosis by analyzing the expression level of signal transducers and activators of transcription (STATS) of apoptotic HEI-OC1 cells line induced by oxidative stress injury.Methods Using different poisoning concentrations (0,20,40,60μM) of(Tert-Butyl hydroperoxide,T-BHP) built the apoptotic HEI-OC1 cells model while 0 μM was used as the control group.The apoptosis level at different poisoning concentration groups was detected by Annexin V-FITC/PI and the expression level of STATs mRNA by real-time quantitative PCR detecting system.Results The apoptosis rates in different contamination groups(0,20,40,60 μM) were 3.9％,7.4％,32.0％,and 91.2％,respectively.Compared with control group,STAT1 mRNA,STAT3 mRNA,STATS mRNA,and STAT6 mRNA expression levels in different poisoning concentration groups declined significantly (F=5 534.302,P＜0.01;F=146.038,P＜0.01;F=685.929,P＜0.01;F=516.11,P＜ 0.01).No statistically significant differences were found among the poisoning concentration groups (P＞0.05).Compared with the control group,the STAT2 mRNA expression levels changed significantly (F=1 259.148,P＜ 0.01).The STAT2 mRNA expression level in 20 μM contamination group decreased while 40,60 μM contamination group increased significantly (P＜0.01).No expression of STAT4 in HEI-OC1 cell was noted.Conclusion The JAK-STAT2 signal pathway has the biological effects on leading oxidative stress injury apoptosis and JAK-STAT1/STAT3/STAT5/STAT6 has the biological effect of inhibit oxidative stress injury apoptosis.

Objective:To establish the reference values and neurological intervention cutoffs for cerebral ventricular size in neonates born at 33 +0-41 +6 weeks of gestation and to investigate the influential factors and reliability of the related indices. Methods:This study prospectively recruited 1 370 1-to 7-day neonates born or hospitalized at the Hunan Provincial Maternal and Child Health Care Hospital from February to August 2021. All the neonates, who were born between 33 +0 and 41 +6 weeks of gestation, were subjected to ultrasound scanning to obtain the indices, including ventricular index (VI), anterior horn width (AHW), thalamo-occipital distance (TOD), and ventricular height (VH). The reference value and neurological intervention cutoff for each index were set. Quantile regression was used to estimate the correlation between each index and continuous covariates [gestational age at birth (GA) and birth weight (BW)]. Mann-Whitney U test was used to analyze the differences in the medians of indices in different categorical covariates groups (males/females, left/right lateral ventricles, vaginal delivery/cesarean section, and singleton/multiple births). Intraclass correlation coefficient (ICC) calculated by a two-way mixed effect model and absolute agreement was used to access intra-rater reliability; ICC via a two-way random effect model and absolute agreement was utilized to rate inter-rater reliability (pool reliability: ICC below 0.50; moderate reliability: ICC between 0.50 and 0.75; good reliability: ICC between 0.75 and 0.90; excellent reliability: ICC exceeding 0.90). Results:The upper limits of reference values for AHW, TOD, VI, and VH in 555 (40.5%) preterm neonates were 2.7-3.5 mm, 20.9-22.5 mm, 12.6-13.7 mm, and 3.8-4.9 mm, and in 815 (59.5%) term newborns were 3.4-4.3 mm, 18.6-21.3 mm, 14.2-14.7 mm, and 3.4-3.8 mm, respectively. The cutoff of neurosurgical intervention for each index was the upper limit of reference value plus 4 mm. AHW median was positively correlated with GA [partial regression coefficient (PRC): 0.12, P<0.05], while TOD and VH medians were negatively correlated with GA (PRC:－0.31 and－0.06, both P<0.05). VI, AHW, and TOD medians were positively associated with BW (PRC: 0.46, 0.23, and 0.97, all P<0.05). The medians of VH, AHW, and TOD in the left cerebral ventricular exceeded those in the right cerebral ventricular, respectively (VH: 2.0 vs 1.8 mm, U=836 071.50; AHW: 1.8 vs 1.7 mm, U=874 141.50; TOD: 13.6 vs 12.5 mm, U=738 409.00, all P<0.05). The medians of AHW and VI in male neonates were greater than those in female newborns, respectively (AHW: 1.8 vs 1.7 mm, U=834 124.00; VI: 11.1 vs 10.8 mm, U=884 156.50, both P<0.05). The neonates delivered vaginally had greater AHW median, but smaller TOD median than those delivered by cesarean section (AHW: 2.0 vs 1.6 mm, U=685 546.00, P<0.001; TOD: 13.1 vs 12.9 mm, U=850 797.00, P=0.010). The AHW median in singleton newborns exceeded that in multiple births (1.9 vs 1.4 mm, U=356 999.00, P<0.001). The lower limits of 95% confidence intervals for intra-rater and inter-rater ICCs exceeded 0.75 and 0.50, respectively. Conclusion:Reference values and surgical intervention thresholds for VI, AHW, TOD, VH of newborns with a gestational age of 33 +0-41 +6 weeks were preliminarily established, and the reliability of these indicators were verified.

Objective:To objectively evaluate the therapeutic effect of Chinese cupping therapy.Method:Near-infrared spectroscopy (NIRS) was used to evaluate the concentration change in oxygenated hemoglobin ([HbO 2]), deoxy-hemoglobin ([Hb]) and blood volume ([tHb]) in 13 healthy volunteers during cupping therapy for the infraspinatus muscle. Results:During the cupping therapy and its pre- and post-treatment period, it was observed that [Hb] and [tHb] in the tissue surrounding the cupping site decreased significantly (both P<0.05), and [HbO 2] increased significantly ( P<0.05), which showed an increase in oxygen uptake. After the cupping was removed, the above hemodynamic changes could still continue for a period of time. Conclusions:Cupping therapy has potential positive therapeutic effects in promoting the hemodynamics for facilitating muscular functions. Functional NIRS surgery has great application potential in the field of physical rehabilitation therapy such as cupping.