Objective:To investigate the short-term efficacy, safety and factors affecting recurrence of intermediate-risk and high-risk non-muscle invasive bladder cancer (NMIBC) treated with intravesical instillation of domestic Bacillus Calmette-Guérin (BCG) infusion.Methods:This study was a retrospective cohort study. We collected the data of 163 patients with NMIBC treated with domestic BCG after transurethral resection of bladder tumor (TURBT) from October 2016 to October 2020 in the Department of Urology, the Affiliated Hospital of Qingdao University. There were 140 males and 23 females, the age was(67.3±10.3) years old. 23 cases had only been received BCG and 140 cases received other chemotherapy drugs before. The induction scheme of instillation was started after the TURBT at once a week for 6 consecutive weeks, continue instillation at every two weeks for 3 doses, then maintenance instillation once a month for 10 consecutive times, for a total of 19 instillations. Kaplan-Meier analysis was used to calculate recurrence-free survival. Binary Logistic regression was used to analyze factors and stepwise regression (backward method) was employed to identify independent risk factors for recurrence after BCG instillation. The incidence of adverse reactions was recorded. Measurement data with normal distribution were expressed as mean±standard deviation( ± s), measurement data with skewed distribution were expressed as M( Q1, Q3), and count data were expressed as frequency and percentage(%). Results:A total of 23 cases experienced recurrence within the 13-month of instillation, of which 7 cases were found to have progressed by pathological biopsy, and the cumulative recurrence-free rate was 85.9%. The results of binary logistic regression analysis showed that the history of re-TURBT ( P=0.010) was an independent predictor for recurrence after BCG intravesical instillation. Adverse events occurred in 110 cases. The main symptoms of the 65 cases were urgency of urination, pollakiuria and dysuria with urinary irritation, urinary tract infection in 15 cases, hematuria in 10 cases, and other symptoms in 20 cases. Instillation was terminated in 7 cases due to side effects, no serious adverse events such as spread of tuberculous bacteria were observed in the cases. Conclusions:Patients with NMIBC treated with intravesical instillation of domestic BCG have significant short-term efficacy, while patients who have previously received re-TURBT is an independent predictor for recurrence. The domestic BCG have the characteristic of slight side effect, good efficacy and safety.

Objective:A recombinant adenoviral vector vaccine based on non-replicating human adenovirus type 5 (Ad5), encoding the conserved domain of respiratory syncytial virus G protein (RSV-G) was constructed. The immunogenicity and protective efficacy of this vaccine were subsequently evaluated in mice.Methods:The recombinant Ad5 vector plasmid (Ad5-Gbcc-Gacc) was constructed by inserted conserved domains of RSV A and RSV B. The recombinant adenovirus Ad5-Gbcc-Gacc was rescued in HEK293A cells. The genome of virus Ad5-Gbcc-Gacc was identified by multi-enzyme digestion, and the expression of Ad5-Gbcc-Gacc was verified by Western blot. Recombinant adenovirus was used to immunize BALB/c mice via intramuscular injection with signal dose, and then challenged with RSV Long strain at week 6. The levels of G specific IgG and antibody subtypes in serum were detected by enzyme-linked immunosorbent assay, the level of neutralizing antibodies was determined by micro-neutralization assay. After challenge, the mice′s weight was recorded daily, the copies of RSV virus in the lung and nasal tissues were detected. Pathological changes in lung tissue were also examined.Results:Western blot and multi-enzyme digestion identification confirmed the successful rescue of the recombinant adenovirus. Ad5-Gbcc-Gacc elicit high titers of specific IgG, robust neutralizing antibodies, and a balanced Th1/Th2 immune response in mice. In comparison to unimmunized controls, mice immunized with Ad5-Gbcc-Gacc reduced the viral copies in both lung and nasal tissue, and exhibited only minimal pathological damage of lung tissue following RSV challenge. In conclusion, Ad5-Gbcc-Gacc induced robust immunogenicity and offers protective effects against RSV infection in murine models.Conclusions:Ad5-Gbcc-Gacc induce robust immunogenicity and can protect mice from RSV challenge, which lays a foundation for further development of RSV vaccine based on G protein.

Objective To investigate the effect of the aqueous extract of Chuan Xiong Rhizoma(CR)on brain metastasis of melanoma B16F10 cells in mice.Methods C57BL/6J mouse models of brain metastasis of melanoma were established by ultrasound-guided intraventricular injection of Luc-labeled B16F10 cells,and brain tumor growth was monitored by in vivo imaging.The mouse models were then randomized for daily gavage of saline or aqueous extract of CR(equivalent crude drug concentration of 1 mg/g).Behavioral tests were used to evaluate the neuroprotective effects of CR in the tumor-bearing mice,and the changes in proteins associated with blood-brain barrier integrity,neuronal cell proliferation and apoptosis,and microglial cell apoptosis and activation were observed using immunofluorescence assay.The efficacy of CR combined with temozolomide(25 mg/kg)against brain metastases of B16F10 cells was observed by in vivo imaging.Results CR-treated mouse models did not show obvious progression of brain metastases and had a reduced rate of body weight loss and lowered protein expressions of ZO-1,claudin-5,occludin,P-gp,TNF-α,AQP4 and PDGFRβ.In the behavioral tests,the CR-treated mice showed prolonged stay on the wooden stick with a shortened time of sticky stick removal.Immunofluorescence assay showed increased proliferation and decreased apoptosis of neuronal cells and microglia in CR-treated mice.CR treatment significantly increased the levels of CD86,CD206,IL-4 and IL-10 and decreased the levels of CD163 and IL-1β in the microenvironment of brain metastases.The mice receiving combined treatments with CR and temozolomide showed significantly lower intensity of fluorescent signals in the brain than those treated with temozolomide alone.Conclusion CR does not promote brain metastasis of melanoma while inducing opening of the blood-brain barrier,and its combined use with TMZ results in enhanced inhibition against brain metastasis of melanoma B16F10 cells in mice.

Tumor metastasis is closely related to high mortality rate of cancer.It is well known that glutamine plays an important role in the malignant progression of cancer.Notably,as an important carbon and nitrogen donor,glutamine has been found to be closely related to tumor metastasis in recent years.Glutamine is not only involved in regulating the proliferation of tumor cells,but is also closely related to the migration and invasion of tumor cells.Furthermore,various enzymes along with transporters in the metabolism of glutamine are involved in the process of tumor metastasis through different signaling pathways.This review provided a summary of the role of glutamine in tumor metastasis in recent years and proposed therapeutic targets to provide new strategies for the clinical treatment of tumor metastases.

Objective To investigate the effect of the aqueous extract of Chuan Xiong Rhizoma(CR)on brain metastasis of melanoma B16F10 cells in mice.Methods C57BL/6J mouse models of brain metastasis of melanoma were established by ultrasound-guided intraventricular injection of Luc-labeled B16F10 cells,and brain tumor growth was monitored by in vivo imaging.The mouse models were then randomized for daily gavage of saline or aqueous extract of CR(equivalent crude drug concentration of 1 mg/g).Behavioral tests were used to evaluate the neuroprotective effects of CR in the tumor-bearing mice,and the changes in proteins associated with blood-brain barrier integrity,neuronal cell proliferation and apoptosis,and microglial cell apoptosis and activation were observed using immunofluorescence assay.The efficacy of CR combined with temozolomide(25 mg/kg)against brain metastases of B16F10 cells was observed by in vivo imaging.Results CR-treated mouse models did not show obvious progression of brain metastases and had a reduced rate of body weight loss and lowered protein expressions of ZO-1,claudin-5,occludin,P-gp,TNF-α,AQP4 and PDGFRβ.In the behavioral tests,the CR-treated mice showed prolonged stay on the wooden stick with a shortened time of sticky stick removal.Immunofluorescence assay showed increased proliferation and decreased apoptosis of neuronal cells and microglia in CR-treated mice.CR treatment significantly increased the levels of CD86,CD206,IL-4 and IL-10 and decreased the levels of CD163 and IL-1β in the microenvironment of brain metastases.The mice receiving combined treatments with CR and temozolomide showed significantly lower intensity of fluorescent signals in the brain than those treated with temozolomide alone.Conclusion CR does not promote brain metastasis of melanoma while inducing opening of the blood-brain barrier,and its combined use with TMZ results in enhanced inhibition against brain metastasis of melanoma B16F10 cells in mice.

Objective: To investigate the change in intestinal flora in Mongolian students with anxiety,so as to provide basis for exploring the relationship between flora and secretion expression in vivo. Methods: The Self rating Anxiety Scale(SAS)was used to assess anxiety in medical college students; then a semi structured interview was administered. Fecal samples that met the inclusion criteria were collected and divided into anxiety (SAS score≥50) and control groups (no anxiety, SAS score<50) according to the standard score of SAS. Samples provided by Mongolian female students were selected from each group. The total bacterial DNA was extracted from the fecal samples for PCR amplification and NovaSeq 2x250bp high throughput sequencing was performed for the V3- V4 region of 16S rDNA gene to obtain the biological information of the intestinal flora. The intergroup OTU, structural diversity, significant difference, and LEfSe analyses were performed with information mining of the literature think tanks. Results: Anxiety existed in 23.86% of the Mongolian students,and 16.96% of the Han people. A Chi square test showed no significant difference in detection of anxiety between Mongolian and Han college students ( P =0.07). Analysis of the alpha diversity index showed that the Shannon index, Simpson index, Chao1 index, and Observed species did not differed significantly between the two groups( t =8.0， 9.0 ，6.0，6.5). The difference in abundance of some bacteria was significant at the Class, Order, Family, and Genus levels between the two groups( t =-2.26-2.57，-5.08-3.58，-2.65-2.09， P <0.05). Conclusion The alpha diversity index showed that there was no significant difference in the abundance and diversity of intestinal flora between the two groups. While there were significant differences at different classification levels, the results suggest that the structure of intestinal flora can change in students with anxiety.

Objective:To study combined adjuvant transcatheter arterial chemoembolization (TACE) with anti-tumor drug treatment on early hepatocellular carcinoma (HCC) recurrence in patients with microvascular invasion (MVI) after partial hepatectomy with curative intent.Methods:The clinical and pathological data of 169 patients with HCC who underwent partial hepatectomy with curative intent from January 2015 to December 2018 at the Affiliated Hospital of Qingdao University were retrospectively analyzed. MVI was diagnosed by postoperative histopathology. There were 147 males and 22 females, with the median age 56 years(ranged 32-79 years). The patients were divided into surgery group ( n=62, patients who did not receive adjuvant therapy), TACE group ( n=42, patients who only received TACE) and combined group ( n=65, patients who received TACE with anti-tumor drug) according to the therapies after resection. Patients in each group were further divided into grade M1 (mild) and grade M2 (severe) subgroups according to the severity of MVI. All patients were followed-up for observing tumor recurrence. The relapse-free survival in the three groups were compared using the Kaplan-Meier method and the log-rank test was used to compare the tumor-free survival rates. Results:The tumor-free survival rates of 169 patients at 1 and 2 years after operation were 59.2% and 40.8%. The tumor-free survival rates at 1 and 2 years after operation were 45.2% and 25.8% in surgery group, 61.9% and 40.5% in TACE group, 70.8% and 52.3% in combined group respectively. The differences among the three groups were significant: TACE group was better than surgery group, and combined group was better than TACE group, combined group was better than surgery group (all P<0.05). In TACE group and combined group, tumor-free survival rates of M1patients better than M2 patients, and the difference was significant ( P<0.05). Among M1 patients and M2 patients, tumor-free survival rates of combined group patients were better than surgery group and TACE group, the difference was significant (all P<0.05). The cumulative tumor-free survival rate was not significantly affected by different antineoplastic agents. Conclusion:Adjuvant TACE reduced the early recurrence rate of HCC patients with MVI. Adjuvant TACE combined with anti-tumor drug further reduced early tumor recurrence.

Objective Investigate the relationship between gross tumor volume (GTV)-related factors including GTV-T volume,the maximum thickness of the esophageal lesion plane and GTV-T volume/length(GTV-T volume divided by the length of the lesion calculated by the number of GTV-T layers) and the locoregional failure of radical intensity-modulated radiation therapy (IMRT) for esophageal carcinoma.Methods A total of 133 patients with esophageal cancer undergoing radical IMRT were enrolled.The factors related to GTV-T including GTV-T volume,the maximum thickness of the esophageal lesions,GTV-T volume/length were calculated.The relationship between GTV-T related factors and local recurrence of tumors was retrospectively analyzed.Results There was positively linear association between the locoregional failure rate of GTV-T and the volume of GTV-T.The volume of GTV-T tumor was 36 cm3,the maximum wall thickness was 2.5 cm,and the GTV-T volume/length was calculated as 5.3 cm2.These critical values could be utilized to predict the risk of locoregional failure of IMRT for esophageal carcinoma.Conclusions The GTV-T factors can be adopted to predict the local control and the risk of locoregional failure of radical IMRT for esophageal carcinoma to certain extent.

Objective To determine the spectrum of mutations responsible for Phenylalanine hydroxylase (PAH) deficiency on phenylketonuria (PKU) patients of Han Chinese people in the Huaihai region of central China.Methods One hundred and one patients diagnosed with PKU were referred to Xuzhou Maternity and Child Health Care Hospital for genetic counseling/analysis from January 2003 to December 2013.Thirteen exons of PAH gene mutations,as well as their flanking introns,were identified in 202 of chromosomes using polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) and sequencing.Results (1) The spectrum was composed of 24 different mutation types,which had been submitted to the National Center for Biotechnology Information(NCBI) dbSNP databases under accession number SS#2137543837_SS#C2137543860.(2)The most commonly affected region was exon 7 and its flanking introns.The most prevalent mutations were c.728G ＞ A (p.R243Q),followed by c.721C ＞ T (p.R241C),c.1155G ＞ C(p.L385L),c.1068C ＞ A(p.Y356X),c.-71A ＞ C(-71A ＞ C) and c.60 + 62C ＞ T (IVS1 +62C ＞T),accounting for 18.317％,8.416％,4.950％,3.960％,3.465％ and 2.970％ of the mutant chromosomes,respectively.(3)Two novel mutations were identified in PAH gene in PKU patients of Han Chinese people:c.60+62C＞T(IVS1 +62C ＞T) and c.782G ＞T(p.R261L).Conclusions The vast majority of PAH mutations identified corresponded to those observed for the PKU populations in the other regions in China,whereas a few are considerably different from others.The mutational spectrum of PAH gene found in patients with PKU in the Huaihai region exhibit regional association.

Objective To analyze the effects of gross tumor volume(GTV-T)and positive lymph node volume(GTV-LN)on the prognosis of radical concurrent chemoradiotherapy for esophageal squamous cell carcinoma(ESCC). Methods A total of 79 patients with stage N1ESCC undergoing radical radiotherapy in our hospital from 2012 to 2015 were enrolled as subjects. GTV-T and GTV-LN were calculated by the Pinnacle39.0 treatment planning system. The receiver operating characteristic(ROC)curves were used to evaluate the value of the GTV-LN/GTV-T ratio in the prediction of local recurrence(LR)and distant metastasis(DM)of ESCC. Results The median follow-up time was 17.2 months in all patients. The ROC curves were made using the GTV-LN/GTV-T ratio. The optimal cut-off values of GTV-LN/GTV-T ratio for predicting the risk of LR and DM were 0.34 and 0.59, respectively. The statistical analysis revealed that the LR rates were 50% and 8% in patients with GTV-LN/GTV-T ratios of<0.34 and ≥0.34, respectively(P<0.01), while the DM rates were 11% and 43% in patients with GTV-LN/GTV-T ratios of<0.59 and ≥0.59, respectively(P= 0.003). Conclusions The GTV-LN/GTV-T ratio may be a predictor of LR and DM in patients with ESCC. Further studies on the GTV-LN/GTV-T ratio may help to make personalized chemoradiotherapy strategies for patients with ESCC.