AIM:To establish a mouse model of chronic skin fibrosis by combining skin injury with bleomycin(BLM)induction.METHODS:Male SPF-grade BALB/c mice were randomly allocated into five groups(n=10 per group):control(Ctrl),high-dose BLM(BLM-H),medium-dose BLM(BLM-M),low-dose BLM(BLM-L),and BLM-control(BLM-Ctrl).A 6 mm full-thickness skin excision was performed on the dorsal region of mice,followed by subcuta-neous injections of BLM at four points around the wound.Mice in the Ctrl group were injected with saline,whereas the BLM-Ctrl group received injections without skin excision.The wound healing rates and times were assessed statistically.Histopathological alterations in wound tissues were examined using hematoxylin-eosin and Masson's trichrome staining.Enzyme-linked immunosorbent assay(ELISA)was employed to measure matrix metalloproteinases(MMPs),and Western blot analysis was conducted to detect collagen type I(COL I)and COL III expression.RESULTS:Compared to the Ctrl group,wound healing rates were significantly reduced(P<0.05 or P<0.01)and healing times significantly prolonged in BLM-H,BLM-M,and BLM-L groups.Histological analysis indicated significantly delayed epithelialization,thicker der-mis,increased collagen deposition,and heightened inflammatory infiltration in the BLM-H group relative to the Ctrl group(P<0.05 or P<0.01).ELISA revealed significantly elevated expression of MMP-2,MMP-3,and MMP-9 in the BLM-H group compared to controls(P<0.01).Western blot results demonstrated significantly increased COL I and COL III pro-tein levels in the BLM-H group compared to the Ctrl group(P<0.05 or P<0.01).CONCLUSION:A mouse model in-volving a 6 mm full-thickness dorsal skin excision combined with subcutaneous injections of BLM(5 mg/kg)at four perile-sional points daily for 21 consecutive days is suitable for investigating chronic skin fibrosis wounds.

AIM:To establish a mouse model of chronic skin fibrosis by combining skin injury with bleomycin(BLM)induction.METHODS:Male SPF-grade BALB/c mice were randomly allocated into five groups(n=10 per group):control(Ctrl),high-dose BLM(BLM-H),medium-dose BLM(BLM-M),low-dose BLM(BLM-L),and BLM-control(BLM-Ctrl).A 6 mm full-thickness skin excision was performed on the dorsal region of mice,followed by subcuta-neous injections of BLM at four points around the wound.Mice in the Ctrl group were injected with saline,whereas the BLM-Ctrl group received injections without skin excision.The wound healing rates and times were assessed statistically.Histopathological alterations in wound tissues were examined using hematoxylin-eosin and Masson's trichrome staining.Enzyme-linked immunosorbent assay(ELISA)was employed to measure matrix metalloproteinases(MMPs),and Western blot analysis was conducted to detect collagen type I(COL I)and COL III expression.RESULTS:Compared to the Ctrl group,wound healing rates were significantly reduced(P<0.05 or P<0.01)and healing times significantly prolonged in BLM-H,BLM-M,and BLM-L groups.Histological analysis indicated significantly delayed epithelialization,thicker der-mis,increased collagen deposition,and heightened inflammatory infiltration in the BLM-H group relative to the Ctrl group(P<0.05 or P<0.01).ELISA revealed significantly elevated expression of MMP-2,MMP-3,and MMP-9 in the BLM-H group compared to controls(P<0.01).Western blot results demonstrated significantly increased COL I and COL III pro-tein levels in the BLM-H group compared to the Ctrl group(P<0.05 or P<0.01).CONCLUSION:A mouse model in-volving a 6 mm full-thickness dorsal skin excision combined with subcutaneous injections of BLM(5 mg/kg)at four perile-sional points daily for 21 consecutive days is suitable for investigating chronic skin fibrosis wounds.

Objective:To investigate the monitoring value of echocardiography for atrial septostomy during the treatment of extracorporeal membrane oxygenation(ECMO).Methods:Sixteen patients underwent atrial septostomy during the treatment of ECMO in Wuhan Asia Heart Hospital from Jan.2017 to Dec.2019 were included. The diameter of the ostomy, the direction of interatrial shunt, the tricuspid regurgitation degree and the pulmonary artery pressure were measured by echocardiography daily after atrial septostomy.Results:There were no significant differences in the diameter of the ostomy in 16 patients(the range of coefficient of variation 0.86%-4.77%). The direction of interatrial shunt changed gradually from right - left atrial shunt to left - right atrial shunt. Pulmonary arterial pressure decreased gradually(the range of coefficient of variation 14.70%-32.13%) as estimated by tricuspid regurgitation pressure gradient.Conclusions:Atrial septostomy is an effective measure to relieve right heart pressure and improve right heart function in patients with ECMO. The echocardiography can be helpful for the monitoring of atrial septostomy.

Objective:To investigate the effect of decreasing DKC1 gene expression on radiosensitivity of HeLa cells.Methods:A cell model with low expression of DKC1 gene was established by shRNA technology with lentivirus as vector, and the interference efficiency was verified by RT-PCR and Western blot assay. Cells were divided into two groups of interference (Lv-shDKC1) and its negative control. Telomerase activity was detected by TRAP-ELISA, and telomere length was measured by Real-time PCR. Cell survival was obtained through clone formation assay and fitted by multi-target single-hit model, and radiobiological parameters ( D0, Dq, N, SF2) and radiosensitization ratio (SER) were calculated. Results:After DKC1 interfering, the expression levels of mRNA and protein of DKC1 in HeLa cells were significantly decreased by (71.330±4.112)% ( t=25.53, P<0.05) and (35.520±3.804)% ( t=4.833, P<0.05), respectively. Compared with the blank control group and negative control group, the telomerase activity of Lv-shDKC1 group decreased significantly from 0.900±0.044 and 0.897±0.031 to 0.713±0.021 ( F=31.44, P<0.05), the relative telomere length was significantly decreased from 4.233±0.306 and 4.633±0.379 to 2.667±0.404 ( F=39.15, P<0.05). The telomerase activity and relative telomere length of blank control group and Lv-shDKC1 negative control group had no significant difference( P>0.05). SF2 in the interference group (0.571±0.006) was significantly lower than that of the blank control group (0.861±0.009) and the Lv-shDKC1 negative control group (0.807±0.002) ( F=1812, P<0.05), and the radiosensitization ratio (SER) of shDKC1 interference was 1.508. Conclusions:Downregulation of DKC1 in human cervical cancer HeLa cells enhances the radiosensitivity through inhibiting the activity of telomerase and shortening the length of telomere. DKC1 gene may become a new target of radiosensitization.

OBJECTIVE：To study the spectrum-ef fect relationship of anti-hepatoma effect of C Ⅱ-3 extract from Periplaneta americana，and to preliminarily clarify the anti-hepatoma active components of it. METHODS ：Based on UHPLC fingerprints of 10 batches of C Ⅱ-3 samples，with the help of UHPL C-Q-TOF/MS，the compounds corresponding to each chromatographic peak were identified qualitatively by standard substances and related literatures. Using the IC 50 value of each batch of C Ⅱ-3 sample against human hepatoma cells HepG 2 as anti-hepatoma activity index ，the spectrum-effect relationship of fingerprint and anti-hepatoma effect was established and analyzed by the combination of grey relational analysis （GRA）and orthogonal partial least squares（OPLS）. RESULTS ：There were 25 common peaks in UHPLC fingerprints of 10 batches of C Ⅱ-3 samples，and 10 chemical compounds were identified ，which were cyclo- （Tyr-Pro）（peak 24），cyclo-（Gly-Phe）（peak 15），hypoxanthine（peak 3）， adenine（peak 7），phenylalanine（peak 10），inosine（peak 11），N-acetyldopamine（peak 16），cyclo-（Pro-Ala）（peak 13），2-hydroxy propionyl（peak 22），cyclo-（Pro-Ser）（peak 6）. The IC 50 of 10 batches of C Ⅱ-3 samples to HepG 2 cells was 70.550-200.303 μg/mL. Among 25 common peaks ，the order of GRA correlation （r）of anti-hepatoma activity was peak 20＞23＞24＞15，all of r values were greater than 0.7；the order of variable importance projection （VIP）of OPLS analysis was peak 23＞18＞15＞24＞7＞14＞6＞ 2＞20，all of VIP values were greater than 1. The standard regression coefficients of peak 7，15，20，23，24 were all greater than 0；while the standard regression coefficients of peak 2，6，14，18 were all less than 0. Conjoint analysis shows that the order of anti-hepatoma activity was peak 20＞23＞24＞15. CONCLUSIONS：unknown chemical ingredients （peak 20， ），cyclo-（Tyr-Pro）（peak 24）and cyclo- （Gly-Phe）（peak 15） in C Ⅱ-3 may be main anti-hepatoma active components.

Objective: To investigate the relationship between the expression of mucin 1 and collagen Ⅳ and the clinical characteristics in invasive breast cancer by using the quantum dots immunofluorescence imaging technique. Methods: The expressions of mucin 1 and collagen Ⅳ were detected simultaneously by quantum dots immunofluorescence imaging technique in 94 cases of breast cancer from July 2007 to July 2010 in Affiliated Hospital of Guilin Medical University. The correlation of mucin 1 and collagen Ⅳ quantitative parameters with clinicopathological features and the prognosis were also analyzed. Results: The positive rates of mucin 1 in human breast cancer tissues marked by quantum dots immunofluorescence imaging technique and immunohistochemistry were 73.4 % (69/94) and 69.1 % (65/94), the positive rates of collagen Ⅳ were 53.2 % (50/94) and 47.9 % (45/94), and the differences were not statistically significant (both P > 0.05) Quantum dots immunofluorescence imaging technique was consistent with conventional immunohistochemistry (IHC) in detecting the expressions of mucin 1 and collagen Ⅳ in human breast cancer tissues (κ = 0.763, P = 0.000; κ=0.759, P = 0.000). The expression of mucin 1 was negatively correlated with collagen Ⅳ (r = -0.883, P < 0.01). The expressions of mucin 1 and collagen Ⅳ were respectively associated with tumor size (F = 3.683, P = 0.029; F = 4.922, P = 0.009), histological grading (F = 3.529, P = 0.033; F = 3.912, P = 0.023), lymph node metastasis (t = -4.868, P = 0.000; t = 3.868, P = 0.000), pathological stage (t = -8.337, P = 0.000; t = 5.962, P = 0.000) and 5-year disease free survival rate (both P = 0.000), and the differences were statistically significant (all P < 0.05). Conclusion The co-detection of mucin 1 and collagen Ⅳ by using quantum dots immunofluorescence imaging technique provides direct evidence determining the biologic behaviors of breast cancer and evaluating the prognosis.

Objective To discuss the diagnostic value of HIT-antibodies in suspected HIT patients with heart diseases.Methods A single center study.We collected 242 blood samples of suspected HIT patients whose platelet count decreased after heparin application during July 1 st ,2012 to June 30th ,2016 in Wuhan Asia Heart Hospital and detected the concentration of HIT antibodies , meanwhile the 4T′s score were calculated.Among the study objects , there are 206 patients received cardiac surgery , 28 received cardiac interventional therapy and 8 received drug therapy.And we divided them into HIT group (44, median age 57.5, 23 females ) and non-HIT group ( 198, median age 63.5, 87 females ) according to clinical diagnosis.Quantitative data was analyzed by independent t-test or Mann-Whitney U test.Qualitative data was analyzed by Fisher′s exact test.We drew ROC curve according to the statistical analysis to determine the optimal threshold value of antibodies in diagnosis of HIT andsensitivity , specificity, negative likelihood ratio, positive likelihood ratio of the HIT antibody detection .Therefore, we can assess the value of HIT antibody detection in HIT clinical diagnosis and treatment .Moreover, we used the optimal threshold value of antibodies to testify the suspected HIT patients .Results The HIT antibody concentration of HIT group (44) and non-HIT group ( 198 ) are 3.2 ( 95% CI:1.8 -5.5 ) U/ml and 0.4 ( 95% CI:0.3 -0.4 ) U/ml, respectively.The concentration of HIT group is much higher than the non-HIT group(P<0.000).When the cut-off value of HIT-Ab is set at 0.9 U/ml, sensitivity and specificity are 93.2%and 91.9%, respectively. And negative likelihood ratio and positive likelihood ratio are 0.07 and 11.53, respectively.When the cut-off value of HIT-Ab is set at 0.6 U/ml, sensitivity and specificity are 100.0%and 73.7%.HIT-Ab and 4T′s score of ROC-AUC are 0.971 and 0.745, respectively.The diagnosis value of HIT-Ab in HIT is significantly higher than the 4T′s score ( P<0.000).Conclusions HIT antibody detection is a simple and effective auxiliary diagnostic method in HIT exclusion .And HIT antibody detection is more optimal than the 4T′s score in HIT diagnosis and treatment .

Objective To understand the status quo of dyslipidemia among urban and rural residents in Ganzhou City.Methods The multistage stratified cluster random sampling method was adopted to select the urban and rural residents aged more than 35 years old as the respondents,among them 4 829 respondents had the intact data,the investigation items included age,sex,height,body mass,waist circumference(WC),FBG,Scr,blood pressure,TC,TG,HDL and LDL.The body mass index was calculated.Results The prevalence rates of hypercholesterolemia,high TG and low HDL among the urban and rural residents in Ganzhou City were 8.76 ％,16.38％ and 24.04％ respectively.Compared with the persons of normal blood lipid,FBG,Ser,WC and BMI in the persons of high blood lipid were much higher (P＜0.05).Conclsion The blood lipid abnormality in the urban and rural residents in Ganzhou City is dominated by low HDL and high TG.

Objective To study the clinical effect of enalapril maleate and folic acid tablets in the treatment of H type hypertension, and to observe the blood homocysteine(Hcy), to provide the basis for clinical diagnosis and treatment.Methods 126 patients with H type hypertension were selected.They were divided into study group and control group according to the random number table method,63 cases in each group.The control group was given enalapril maleate, the study group was given enalapril maleate and folic acid tablets.The clinical efficacy of the two groups was compared.The systolic blood pressure, diastolic blood pressure and Hcy level before and after treatment in two groups were observed.Results The total effective rate of the study group was 90.5%(57/63),which of the control group was 74.6%(47/63),the difference was statistically significant(x2=15.371,P<0.05).There was no significant difference in systolic blood pressure and diastolic blood pressure before treatment between the two groups(P>0.05).The systolic pressure after treatment of the two groups were (129.3±5.7)mmHg and (137.9±4.9)mmHg, the diastolic blood pressure were (81.5±3.6)mmHg and (89.6±5.1)mmHg, the differences were statistically significant compared with before treatment(t=8.969,7.851,7.343,6.987,all P<0.05),the difference between the two groups was statistically significant(P<0.05).There was no significant difference in Hcy before treatment between the two groups(P>0.05), after treatment Hcy levels of the two groups were (8.93±0.63)μmol/L and (15.35±0.57)μmol/L respectively, the differences were statistically significant compared with before treatment(t=8.097,8.031,all P<0.05),the difference between the two groups was statistically significant(t=7.573,P<0.05).Conclusion Enalapril maleate and folic acid tablets in the treatment of H type hypertension has good clinical effect, can significantly lower blood pressure, improve Hcy level.

DNA microarray has become an essential medical genetic diagnostic tool for its high-throughput, miniaturization and automation. The design and selection of oligonucleotide probes are critical for preparing gene chips with high quality. Several sets of probe design software have been developed and are available to perform this work now. Every set of the software aims to different target sequences and shows different advantages and limitations. In this article, the research and development of these sets of software are reviewed in line with three main criteria, including specificity, sensitivity and melting temperature (Tm). In addition, based on the experimental results from literatures, these sets of software are classified according to their applications. This review will be helpful for users to choose an appropriate probe-design software. It will also reduce the costs of microarrays, improve the application efficiency of microarrays, and promote both the research and development (R&D) and commercialization of high-performance probe design software.
Base Sequence ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; Sensitivity and Specificity ; Software ; Software Design