Objective To establish a real-time PCR based method for detection of survivin gene samples methylation with dye SYBR Green Ⅰ.Methods DNA samples from 25 pairs of gastric cancer tissue and matched normal gastric tissues were digested with mCpG-sensitive Hpa Ⅱand Msp Ⅰ, and detected by SYBR Green Ⅰ real-time PCR for survivin exon 1 specific primers. The concentration of digested DNA was monitored by real-time quantitative PCR with survivin intron 2 specific primers. A normal genomic DNA which served as a standard was diluted serially 10-fold to determine the sensitivity of real-time PCR. The products were further verified by agarose gel electrophoresis analysis.Results After HpaⅡ enzyme digestion, a peak of PCR product derived from methylated targeted gene was found in the melting curve at Tm (91.5±0.5) ℃, which was verified as 338 bp strap. The control gene (survivin intron 2) characteristic of Tm (79.5±0.5) ℃ was amplified in all digested samples, indicating no non-specific degradation occurred in the genome DNA. The sensitivity of this method was 100 copies/μL. The new established PCR system was applied to detecting 25 cases of lung cancer tissue samples. As a result, the frequency of survivin exon 1 demethylation was 96%.Conclusion As a reliable new method for detection of survivin exon 1 methylation, SYBR Green Ⅰ real-time fluorescent PCR assay is rapid, accurate, sensitive, timing and simple.

Objective To assess the efficacy of high intensity focused ultrasound (HIFU) for the ablation of feeding-artery in uterine fibroids.Methods A total of 45 women with uterine fibroids who had children accepted HIFU treatment.Color Doppler and contrast-enhanced ultrasound were used to target the supply vessels entered the fibroid.HIFU treatment was focused primarily on locations where supplying vessels entered the fibroid.Conventional ultrasound,color Doppler ultrasound,contrast-enhanced ultrasound and MRI were performed before and after HIFU treatment to observe the changes of imaging.Results After HIFU treatment,the volume of uterine fibroids decreased to certain extent.The blood signals decreased or disappeared in the treated regions.Bewteen two groups of the uterine fibroids diamete ≥5 cm and ＜5 cm,there were significant differences in the percentage of the nonperfusion regions in the volume of uterine fibroids (≥70％,P ＜0.05).Conclusions Ablation of feeding artery by HIFU is an effective treatment for the uterine fibroids.HIFU treatment is effective for the ablation of feeding-artery in uterine fibroids whose diameter less than 5 cm.

Survivin variants specific real time quantitative RT-PCR was developed to analyze their expression in 53 paired cancer and para-cancerous tissues, and the expression of the wild-type survivin protein was detected by immunohistochemistry. The results showed that survivin mRNA and protein were expressed in gastric cancer and para-cancerous tissues. The survivin-2B was dominantly expressed in para-cancerous tissues, whereas the survivin-DeltaEx3 was more frequently detected in cancer tissues. The positive rate of survivin-2a was 100% in both cancer and para-cancerous tissues, but its relative transcript expression level was not significantly increased in cancer tissues in comparison with para-cancerous tissues. The correlation analysis revealed that the expression of survivin-2a mRNA was significantly associated with that of total survivin (r (s)=0.4178, P=0.0018), whereas inversely to that of survivin-DeltaEX3 (r (s)=-0.4506, P=0.0007). It was suggested that survivin-2a may act as an antagonist of survivin-DeltaEX3. The balance between antiapoptotic survivin iso-forms and nonantiapoptotic ones may play an important role in tumorigenesis and tumor progression. Promising value is hinted to analyze survivin and its variants in tumor early diagnosis and distinguishing malignant tumors from benign ones.
Microtubule-Associated Proteins/genetics ; Microtubule-Associated Proteins/*metabolism ; Protein Isoforms/genetics ; Protein Isoforms/metabolism ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Stomach Neoplasms/*metabolism ; Stomach Neoplasms/pathology ; Tumor Cells, Cultured ; Tumor Markers, Biological/genetics ; Tumor Markers, Biological/*metabolism

Objective To investigate the correlations of expression of interferon inducible protein 10 ( IP-10) mRNA in peripheral blood mononuclear cells ( PBMCs) with serum levels of HBsAg and HBV DNA in patients with HBV-related acute on chronic liver failure ( HBV-ACLF ) , and to assess its value in predicting disease prognosis .Methods Eighty patients with HBV-ACLF, 60 patients with chronic hepatitis B (CHB), and 25 healthy subjects were enrolled during October 2013 and February 2015.IP-10 mRNA in PBMCs was measured by real time quantitative polymerase chain reaction ( RT-qPCR) , and the difference in IP-10 mRNA expression among three groups was compared by ANOVA .Serum levels of HBsAg and HBV DNA were also measured in patients with HBV-ACLF, and Pearson correlation test was performed to analyze the correlations of IP-10 mRNA with HBsAg and HBV DNA levels .t test was used to analyze the differences in IP-10 mRNA, HBsAg and HBV DNA levels between fatal and surviving cases after 3-month entecavir therapy in HBV-ACLF group.Receiver operating characteristic curves ( ROC) was used to evaluate the prognostic value of IP-10 mRNA, HBsAg and HBV DNA in HBV-ACLF patients .Results IP-10 mRNA level in HBV-ACLF group was 0.998 ±0.186, which was higher than those in CHB patients and healthy controls (0.641 ±0.083 and 0.412 ±0.062, t=3.841 and 16.661, P<0.01).Pearson correlation analysis showed that IP-10 mRNA level was negatively correlated with HBsAg level in HBV-ACLF patients (r=-0.576, P<0.01), but positively correlated with HBV DNA level (r=0.547, P<0.01).After 3-month entecavir therapy , IP-10 mRNA level in surviving cases of HBV-ACLF group was 0.894 ±0.164, which was lower than that in fatal cases ( 1.103 ±0.177, t =-4.328, P <0.01 ); HBsAg level in surviving cases was higher than that in fatal cases (3.303 ±0.565 vs.2.605 ±0.844, t =3.251, P<0.01).The area under ROC of IP-10 mRNA in evaluating prognosis of HBV-ACLF was 0.820, which was higher than those of HBsAg (0.663) and HBV DNA (0.570).Conclusions IP-10 mRNA in patients with HBV-ACLF is over-expressed and is correlated with HBsAg and HBV DNA levels .It may be used for predicting the prognosis of patients with HBV-ACLF.

Long noncoding RNA (lncRNA) is a class of RNA molecules that longer than 200 nucleotides.lncRNA is lack of functional open reading frames and has no protein coding ability.Recently, accumulating evidences indicate that lncRNA plays active roles in tumor carcinogenesis and progression.The aberrant expression of lncRNA is significantly correlated with the growth,metastasis,and therapy-resistance of digestive system tumors.Thus,lncRNA may be served as new targets for the diagnosis,treatment,and progno-sis of patients with digestive system tumors.

Objective To investigate the expression levels of serum LINC00978 in gastric cancer patients and its correlation with clinicopathological characteristics,and evaluate its clinical diagnostic value.Methods The expression levels of LINC00978 in gastric cancer cells and serum samples from the patients with gastric cancer or gastric benign diseases and healthy controls were detected by real-time qRT-PCR,and its correlations with clinicopathological parameters were analyzed.The diagnostic efficacy of serum LINC00978 was evaluated by the receiver operating characteristic (ROC) curve.Results The expression levels of LINC00978 in MGC-803 cells were significantly higher than that in GES-1 cells (18.88 ± 1.15 vs 1.00 ± 0.03,t =-21.926,P ＜ 0.05).The expression levels (median[P25,P75]) of serum LINC00978 in gastric cancer patients (3.525 [1.385,8.954]) were significantly higher than those in the patients with gastric benign diseases (0.419 [0.258,1.369],Z =-4.834,P ＜ 0.01) and healthy controls (0.814 [0.351,2.510],Z =-4.686,P ＜ 0.01).The expression levels of serum LINC00978 in gastric cancer patients were significantly related to lymphatic metastasis (r =0.448,P ＜ 0.01),invasive depth (r =0.369,P ＜ 0.01) and TNM stage (r =0.383,P ＜ 0.01),and those after operation significantly lower than before operation (0.17 [0.15,0.39] vs 0.98 [0.59,1.61],Z =-5.731,P ＜ 0.01).There was no significant difference in serum LINC00978 levels between the patients with gastric benign diseases and healthy controls (Z =-1.693,P ＞ 0.05).The area under the ROC curve (AUCROC) and 95％ confidence interval (CI) of serum LINC00978 levels in the diagnosis of gastric cancer were 0.807 and 0.732-0.882,respectively.When the cutoff value was 1.806,the sensitivity and specificity for the diagnosis of gastric cancer were 71.4％ and 75.9％,respectively.Conclusion The expression levels of serum LINC00978 in gastric cancer patients increase and are related to lymphatic metastasis,invasive depth and TNM stage,which may be served as a novel biomarker for the diagnosis of gastric cancer.

Survivin variants specific real time quantitative RT-PCR was developed to analyze their expression in 53 paired cancer and para-cancerous tissues, and the expression of the wild-type survivin protein was detected by immunohistochemistry. The results showed that survivin mRNA and protein were expressed in gastric cancer and para-cancerous tissues. The survivin-2B was dominantly expressed in para-cancerous tissues, whereas the survivin-△Ex3 was more frequently detected in cancer tissues. The positive rate of survivin-2a was 100% in both cancer and para-cancerous tissues,but its relative transcript expression level was not significantly increased in cancer tissues in comparison with para-cancerous tissues. The correlation analysis revealed that the expression of survivin-2a mRNA was significantly associated with that of total survivin (rs=0.4178, P=0.0018),whereas inversely to that of survivin-/EX3 (rs=-0.4506, P=0.0007). It was suggested that survivin-2a may act as an antagonist of survivin-△EX3. The balance between antiapoptotic survivin iso-forms and nonantiapoptotic ones may play an important role in tumorigenesis and tumor progression. Promising value is hinted to analyze survivin and its variants in tumor early diagnosis and distinguishing malignant tumors from benign ones.

Objective:To explore diagnostic value of magnetic resonance apparent diffusion coefficient(ADC)combined with dynamic enhanced magnetic resonance imaging(DCE-MRI)for parotid lymphoma of parotid gland.Methods:Clinical data of 74 patients with tumors of parotid gland who admitted to Hainan Western Central Hospital from September 2021 to September 2023 were selected to carry out retrospective analysis,and they were divided into malignant tumor group(15 cases)and benign tumor group(59 cases)according to the diagnosis of surgical pathology.And then,the benign tumor group was further divided into two subgroups according to the result of surgical pathology,which included pleomorphic adenoma group(35 cases)and"parotid lymphoma of parotid gland"group(24 cases).The differences in ADC values and DCE-MRI parameters(Ve,Kep,Ktrans)were compared between malignant tumors and benign tumors,and between pleomorphic adenoma and parotid lymphoma of parotid gland.The efficacies of ADC values and DCE-MRI parameters in diagnosing malignant tumors were analyzed by means of the receiver operating characteristics(ROC)curve.Results:The ADC value,Ve value and Kep of benign tumor group were respectively(1.17±0.41)×10-3 mm2/s,(0.46±0.15)and(1.56±0.57)min,which were significantly higher than those[0.86±0.19× 103mm2/s,0.35±0.08,and 0.80±0.27 min]of malignant tumor group(t=2.841,2.733,5.004,P＜0.05),respectively.Results of binary logistic regression analysis showed that ADC,Ve and Kep were respectively influence factors on benign and malignant parotid tumors(OR=0.045,0.001,0.004,P＜0.05).ROC curve analysis showed that the AUC values of single ADC,Ve,Kep,and the combination of them were respectively 0.726,0.715,0.903 and 0.915 in diagnosing benign and malignant parotid tumors.There was not significant difference in Ktrans value between pleomorphic adenoma group and"parotid lymphoma of parotid gland"group(P＞0.05).The ADC value and Ve value of"parotid lymphoma of parotid gland"group were lower than those of pleomorphic adenoma group,while Kep value of"parotid lymphoma of parotid gland"group was higher than that of pleomorphic adenoma group,and the differences of them between two groups were significant(t=4.568,4.897,4.805,P＜0.05),respectively.Binary logistic regression analysis showed that ADC,Ve and Kep were influential factors of occurring parotid lymphoma of parotid gland(OR=0.042,0.000,13.452,P＜0.05),respectively.ROC curve analysis showed that the AUC values of single ADC,Ve and Kep,and the combined diagnosis of them were respectively 0.742,0.784,0.763 and 0.959 for parotid lymphoma of parotid gland.Conclusion:Magnetic resonance ADC value and DCE-MRI parameter have favorable value in identifying benign and malignant parotid tumors,and in identifying and diagnosing parotid lymphoma of parotid gland and pleomorphic adenoma,and the combined diagnosis has higher diagnostic value.

Objective Toinvestigatetheeffectofpressureandnon-pressureonthedirectimagingoflowerlimbdeepveinswith CTvenography(CTV).Methods 100patients(50malesand50females,aged30－80yearsold,mean (63.5±13.5yearsold)with suspectedlowerextremityvenousdiseasesfrom September2015to October2018 wereretrospectivelyanalyzed.50patientswere scannedafterpressingtheankle(controlgroup),andtheother50werescannedwithoutpressingtheankle(experimentalgroup).Results Therewerenosignificantdifferencesbetweenthecontrolandexperimentalgroupsintheauxiliaryveinscore (t＝－0.20,P＝0.82), femoralveinscore(t＝－0.1,P＝0.91),andtotaliliacveinscore(t＝－0.03,P＝0.97).Conclusion CTV withoutpressingtheankle demonstratescomparableefficacytodirectimagingoflowerlimbdeepveins,withgoodimagequality,reducedcomplicationsandeasy toapply,sothatitshouldbewidelyusedinclinicalpractice.

Methods:Whole blood and serum samples were collected from 86 patients with HBV-ACLF and 42 patients with chronic hepatitis B (CHB). Serum alanine aminotransferase (ALT), total bilirubin (TBil), creatinine (Cr), albumin (ALB) cholinesterase (CHE) and international standardized ratio (INR) were detected by routine method . The CXCL10 mRNA level of peripheral blood mononuclear cells was detected by fluorescence quantitative PCR. The genotype and allele of rs56061981 were detected by direct DNA sequencing. The genotype and allele frequencies between the ACLF and CHB groups were compared. The levels of ALT, TBil, INR, ALB, CHE, model for end-stage liver disease (MELD) and CXCL10 mRNA were compared between the two groups according to whether the T allele was carried.Results:There was no significant difference in sex, age, alcohol history, smoking history, HBV genotype, HBeAg status and HBV-DNA level between CHB group and HBV-ACLF group (P > 0.05). The frequency of CT+ TT genotype and the frequency of T allele at the rs56061981 locus of the CXCL10 gene in group ACLF were significantly higher than that in the CHB group ( χ2=4.83, P=0.03 and χ2=4.95, P=0.03). The difference is significant Genotype CT+ TT is a risk factor for CHB to develop into HBV-ACLF ( OR=2.897, 95% CI: 1.09~7.68). Allele T is a risk factor for CHB to develop into HBV-ACLF ( OR=2.746, 95% CI: 1.10~6.89). The plasma INR and MELD scores of ACLF individuals carrying T alleles were significantly higher than those carrying C alleles (t=2.63, P=0.013 and t=2.7, P=0.011). The difference is significant. Albumin (ALB) and cholinesterase (CHE) was significantly lower than those carrying C alleles (t=2.67, P=0.01 and t=3.545, P=0.001). The difference is significant. But there was no significant difference in serum TBL and ALT between the two groups. Conclusions:The polymorphism of CXCL10 gene rs56061981 is significantly correlated with the susceptibility and severity of ACLF. Allele T may be a susceptible gene of ACLF, and it can also be used as a predictor of the severity of the disease.