To prepare a lipid nanoparticle (LNP)-based subunit vaccine of Mycobacterium tuberculosis (Mtb) antigen EsxV and study its immunological characteristics, the LNP containing EsxV and c-di-AMP (EsxV: C: L) was prepared by thin film dispersion method, and its encapsulation rate, LNP morphology, particle size, surface charge and polyphase dispersion index were measured. BALB/c mice were immunized with EsxV: C: L by nasal drops. The levels of serum and mucosal antibodies, transcription and secretion of cytokines in lung and spleen, and the proportion of T cell subsets were detected after immunization. EsxV: C: L LNPs were obtained with uniform size and they were spherical and negatively charged. Compared with EsxV: C immunization, EsxV: C: L mucosal inoculation induced increased sIgA level in respiratory tract mucosa. Levels of IL-2 secreted from spleen and ratios of memory T cells and tissue-resident T cells in mice were also elevated. In conclusion, EsxV: C: L could induce stronger mucosal immunity and memory T cell immune responses, which may provide better protection against Mtb infection.
Animals ; Mice ; Mycobacterium tuberculosis ; Antigens, Bacterial ; Immunization ; Nanoparticles ; Vaccines, Subunit ; Mice, Inbred BALB C

Objective:To understand the etiology and clinical characteristics of hospitalized severe community-acquired pneumonia(SCAP) in Changchun, and provide scientific basis for its etiology diagnosis and targeted treatment.Methods:The study subjects included 618 children with clinical diagnosis of SCAP who were hospitalized from January 2016 to December 2019.We collected pharyngeal swabs and alveolar lavage fluid from children.Virus isolation, bacterial culture, time-of-flight mass spectrometry, PCR/RT-PCR, colloidal gold method and Optochin test were used to detect the antigen, nucleic acid and protein profiles in the specimen.Results:There were more boys than girls in hospitalized children with SCAP.The peak age of onset was 7 to 12 months.Most cases occurred in winter and spring.The highest detection rate of SCAP virus was 56.15%(347/618); 73.49%(255/347) were positive for one virus, among which the top five were respiratory syncytial virus (27.8%), influenza A virus (23.9%), influenza B virus (16.1%), rhinovirus (12.2%) and metapneumovirus (10.2%). Two viruses were positive for 19.88%(69/347); three viruses were positive for 4.32%(15/347); four viruses were positive for 2.31%(8/347). Atypical microbial infections were 29.77%(184/618), of which Mycoplasma pneumoniae accounted for 95.65%(176/184). Bacterial infections were 17.31%(107/618), mainly Streptococcus pneumoniae(39.25%, 42/107) and Staphylococcus aureus(24.30%, 26/107). The mixed infection of multiple pathogens was 7.61%(47/618), among which the mixed infection rates of Mycoplasma pneumonia with Streptococcus pneumoniae, virus were 40.43% and 34.04%, respectively.High fever, faster breathing, and perioral cyanosis were risk factors for SCAP, with OR and 95% CI of 7.71 and 4.56-13.04, 2.43 and 2.02-2.93, 3.53 and 2.56-4.86, respectively.Viral co-infection occurred in 36.96%(34/92) of complications such as heart failure, toxic encephalopathy, and myocardial damage; Mycoplasma pneumoniae and other pathogens co-infected 35.29% of children with pleural effusion. Conclusion:The pathogens of SCAP in Changchun are mainly viruses notably, respiratory syncytial virus is the dominant pathogen, followed by Mycoplasma pneumoniae.The bacterial pathogen is mainly Streptococcus pneumoniae.High fever, faster breathing, and cyanosis around the mouth are risk factors for severe pneumonia.Multi-pathogen mixed infection is prone to serious complications.

Objective:To explore the effect of the resection of the primary lesion on the prognosis for patients with stage Ⅳ breast cancer.Methods:A total of 132 breast cancer patients who were first diagnosed as stage Ⅳ in the Hebei Cancer Hospital from June 2008 to June 2015 were divided into two groups: the primary resection group ( n=85) and the unresection group ( n=47). The influences of primary resection, timing of operation, lymph node removal or dissection and radiotherapy on the prognosis of stage Ⅳ breast cancer patients were analyzed. Results:Multivariate Logistic regression analysis showed that visceral metastasis was an independent influencing factor for primary lesion resection in stage Ⅳ breast cancer patients ( OR=2.590, 95% CI: 1.090-6.159). Multivariate Cox regression analysis showed that primary resection was an independent factor for the improvement of prognosis in stage Ⅳ breast cancer patients ( OR=0.582, 95% CI: 0.400-0.847). The median overall survival (OS) was 37.20 months in the resection group, which was higher than 24.10 months in the unresection group ( χ2=8.108, P=0.004). Among patients aged ≥50 years old, the median OS was 39.30 months in the resection group and 23.03 months in the unresection group, and the difference was statistically significant ( χ2=14.191, P<0.001). The median OS was 38.00 months in the 66 patients with the operation time from diagnosis to resection of primary lesion<6 months ( n=66), and 35.20 months for ≥6 months ( n=19) ( χ2=4.430, P=0.035), the difference was statistically significant ( χ2=4.430, P=0.035). The median OR of axillary lymph node dissection and axillary lymph node excision group were 45.37 months and 33.44 months, respectively, the difference was statistically significant ( χ2=7.832, P=0.005). The median OS of postoperative radiotherapy group and non-radiotherapy group were 44.80 months and 33.20 months, respectively, the difference was not statistically significant ( χ2=2.950, P=0.086). Conclusion:Resection of the primary lesion may prolong the survival time of some advanced breast cancer patients.

Objective:To explore the effect of the resection of the primary lesion on the prognosis for patients with stage Ⅳ breast cancer.Methods:A total of 132 breast cancer patients who were first diagnosed as stage Ⅳ in the Hebei Cancer Hospital from June 2008 to June 2015 were divided into two groups: the primary resection group ( n=85) and the unresection group ( n=47). The influences of primary resection, timing of operation, lymph node removal or dissection and radiotherapy on the prognosis of stage Ⅳ breast cancer patients were analyzed. Results:Multivariate Logistic regression analysis showed that visceral metastasis was an independent influencing factor for primary lesion resection in stage Ⅳ breast cancer patients ( OR=2.590, 95% CI: 1.090-6.159). Multivariate Cox regression analysis showed that primary resection was an independent factor for the improvement of prognosis in stage Ⅳ breast cancer patients ( OR=0.582, 95% CI: 0.400-0.847). The median overall survival (OS) was 37.20 months in the resection group, which was higher than 24.10 months in the unresection group ( χ2=8.108, P=0.004). Among patients aged ≥50 years old, the median OS was 39.30 months in the resection group and 23.03 months in the unresection group, and the difference was statistically significant ( χ2=14.191, P<0.001). The median OS was 38.00 months in the 66 patients with the operation time from diagnosis to resection of primary lesion<6 months ( n=66), and 35.20 months for ≥6 months ( n=19) ( χ2=4.430, P=0.035), the difference was statistically significant ( χ2=4.430, P=0.035). The median OR of axillary lymph node dissection and axillary lymph node excision group were 45.37 months and 33.44 months, respectively, the difference was statistically significant ( χ2=7.832, P=0.005). The median OS of postoperative radiotherapy group and non-radiotherapy group were 44.80 months and 33.20 months, respectively, the difference was not statistically significant ( χ2=2.950, P=0.086). Conclusion:Resection of the primary lesion may prolong the survival time of some advanced breast cancer patients.

BACKGROUND: MicroRNAs (miRNAs) are non-coding small molecule RNAs that are widely found in eukaryotic organisms, although some miRNAs have been found in tumors, the expression and effects of miR-665 on small cell lung cancer (SCLC) are unclear. The aim of this study was to analyze the effects of miR-665 on proliferation, cycle, invasion and migration of SCLC cells, and to explore the role of miR-665 in SCLC and its working mechanism. METHODS: The expression of miR-665 in SCLC tissues and adjacent normal tissues was detected by qRT-PCR. TargetScan predicted potential target genes for miR-665 and validated with dual luciferase reporter assays, qRT-PCR and Western blot. CCK8 assay, flow cytometry, Transwell and wound healing assay to detect the effects of miR-665 and LLGL1 on proliferation, invasion, migration and S-phase fraction of SCLC cell line NCI-H446, NCI-H1688. A nude mouse xenograft model of SCLC was constructed and the effect of miR-665 on tumor growth in mice was observed. RESULTS: The expression of miR-665 in SCLC tissues was significantly higher than that in non-tumor normal tissues. MiR-665 could target 3'-UTR of LLGL1 and inhibit its expression. Compared with non-tumor normal tissues, the expression of LLGL1 was significantly lower in SCLC tissues. Inhibition of miR-665 expression could inhibit proliferation, S-phase fraction, invasion and migration ability of SCLC NCL-H446 cells, and interference LLGL1 expression could reverse this inhibition effect. Up-regulation of miR-665 expression could promoted proliferation, S-phase fraction, invasion and migration ability of SCLC NCI-H1688 cells, but this promotion effect was also reversed by overexpression of LLGL1. In a nude mouse xenograft model of SCLC, inhibition of miR-665 expression could up-regulate LLGL1 protein expression and inhibit tumor growth, while up-regulation of miR-665 expression could produce opposite results. CONCLUSIONS The expression of miR-665 is closely related to SCLC. miR-665 can promote the biological behavior of SCLC cells by inhibiting the expression of target gene LLGL1, and miR-665 play a role in tumor-promoting genes in SCLC.

[Abstract] Objective：To explore the regulatory effect of miR-9 on biological behaviors of small cell lung cancer (SCLC) cells by targeting zinc finger E-box binding homeobox 2 (ZEB2), and to analyze the role of miR-9 in SCLC and its possible mechanism. Methods: qPCR, WB and immunohistochemistry methods were used to detect the mRNA and protein expressions of ZEB2 in cancer tissues and corresponding adjacent tissues of 67 SCLC patients who received surgical treatment at the Department of Oncology, Fourth Hospital of Hebei Medical University from February 2018 to November 2019. TargetScan was used to predict the potential target gene of miR-9, which was later verified by Dual luciferase reporter gene assay, qPCR and WB methods. CCK-8 method, Flow cytometry and Transwell experiment were used to detect the effect of miR-9 and ZEB2 over-expression on the biological behaviors of NCI-H446 cells, and WB was used to detect the protein expressions of E-cadherin, N-cadherin and Vimentin in cells. NCI-H446 cells overexpressing miR-9 were used to construct SCLC nude mouse xenograft model, and the effect of miR-9 on the growth of xenografts was observed. Results: The mRNA and protein expression levels of ZEB2 in SCLC tissues were significantly higher than those in adjacent tissues (P<0.01). There is a potential binding site on the 3' UTR of ZEB2 to bind with miR-9. Compared with the control group, the mRNA and protein expression levels of ZEB2 in NCI-H446 cells of the miR-9 over-expression group were significantly reduced (P<0.01); the proliferation, migration and invasion abilities of NCI-H446 cells were significantly suppressed (P<0.05 or P<0.01), and the expression of EMT protein was reduced; However, simultaneous over-expression of ZEB2 could reverse above effects. In in vivo experiments, the size and weight of transplanted tumors in the miR-9 over-expression group were significantly lower than those in the control group (P<0.05 or P<0.01). The expression of ZEB2 protein in the tumor tissues of nude mice in the miR-9 overexpression group was significantly lower than that in the control group (P<0.01). Conclusion: miR-9 can inhibit the biological behaviors of SCLC cells and the growth of NCI-H446 transplanted tumors in nude mice by targeting and regulating ZEB2.

Objective: To understand the clinical features and pathogenic spectrum of encephalitis and meningitis syndrome in children. Methods: A total of 667 cases of children with encephalitis or meningitis diagnosed and documented at Changchun Children′s Hospital from May 2012 to July 2015 were enrolled.A variety of samples in diffe-rent types were collected and presented, including 335 cerebrospinal fluid specimens, 530 blood samples, and 332 stool samples.All the samples were collected from the patients within 72 hours on admission.Moreover, these samples are analyzed and tested, including PCR for enterovirus(EV), herpesvirus(HSV), mycobacterium tuberculosis(TB) and Mycoplasma pneumoniae(MP) nucleic acid in cerebrospinal fluid samples; fecal specimens were tested for EV, enterovirus 71 (EV71), coxsackievirus A6 (CA6), coxsackievirus A16 (CVA16), coxsackievirus A10 (CVA10) nucleic acids; degenerate primers to amplify Echovirus 30 (Echo30). Clinical data of children were collected. Results: The peak incidence of encephalitis and meningitis syndrome was from June to August, age distribution was from 0 to 15 years old, the proportion of children aged from 0-6 accounted for 81.41%; the highest proportion was among 0-1 years old infants, occupying 32.38%; 408 males and 259 females; the main symptoms were fever(586 cases), apathy(337 cases), vomiting (307 cases) and headache(203 cases). And clinical signs included drowsiness (103 cases), neck stiffness (71 cases), meningeal irritation (12 cases), and pathological reflex (313 cases), etc.The clinical diagnosis included 272 cases of viral encephalitis, 332 cases of severe hand, foot and mouth disease complicated by encephalitis, 30 cases of bacterial meningitis, and 33 other cases; the etiological detection included: the positive rates of EV, EBV and Echo30 in cerebrospinal fluid specimens were 59.72%, 3.16% and 70.00%, respectively.And EV71, CVA16, CVA6, EV71+ CA16 and EV71+ CVA16+ CVA6 nucleic acids were detected in fecal samples, in which the highest detection rate was EV71(98.96%). Conclusions In Changchun Children′s Hospital, the children with encephalitis and meningitis are mainly viral encephalitis.The main symptoms were fever, apathetic, drowsiness, vomiting and headache.Signs included, neck stiffness, meningeal irritation, and pathological reflexes, etc.The main pathogen of the disease is EV71.

Objective: To detect the expression of miR-133a-3p in gastric cancer (GC) tissues and plasma of GC patients, and to investigate its effect on the proliferation of GC cells as well as its correlation toprognosis of GC patients. Methods: 52 cases of cancertissues (non-necrosis part) and corresponding adjacent tissues as well as the pre-operative peripheral blood samples from GC patients, who underwent surgery at Department of General Surgery, the Forth Hospital of Hebei Medical University(Shijiazhuang, China) between May 2012 and May 2013, were collected for this study. The plasma sample (n=35) from healthy donors were obtained during their physical examination. RT-qPCR was adopted to detect the expression of miR-133a-3p in gastric cancer tissues, adjacent tissuesand plasma samples of GC patients and healthy volunteers. The relationships between miR-133a-3p expression and the median DFS as well as clinicopathological parameters were also analyzed. CCK-8 assay was adopted to detect the effect of miR-133a-3p silence or over-expression on proliferation of gastric cancer SGC7901 cells. Results: miR-133a-3p was dramatically decreased in gastric cancer tissues (P<0.01), and its expression was associated with TNM stage, tumor infiltration (T), lynphonode metastasis (N), and vascular tumor thrombus (all P<0.01); miR-133a-3p was significantly increased in the plasma of GC patients (P<0.01), and its expression was associated with TNM stage, lynphonode metastasis (N), and vascular tumor thrombus (all P<0.05). miR-133a-3p expression was positively correlated with serum CA199 level of GC patients (P<0.01). The median DFS of patients with high miR-133a-3pexpression in cancer tissues was significantly longer than that of the patients with low expression(20.8 vs 14.8 months, P<0.05); The median DFS of patients with high plasma miR-133a-3p expression was significantly shorter than that of the patients with low expression (14.4 vs 20.3 months, P<0.05). Over-expression of miR-133a-3p could significantly inhibit the proliferation of gastric cancer SGC7901 cells, while miR-133a-3p silence could significantly promote the proliferation (all P<0.05). Conclusion: miR-133a-3p could significantlyinhibit the proliferation of SGC7901 cells; miR-133a-3p aberrantlyexpressed in gastric cancer tissues and plasma, and obviously correlated with prognosis of gastric cancer patients, which may be used as a potential clinical bio-maker for early diagnosis and treatment as well as the prognosis prediction of gastric cancer.

Objective: To investigate the safety, efficacy and prognostic factors of S-1 versus capecitabine in patients with advanced breast cancer (ABC). Methods: From January 1, 2012 to December 31, 2014, 154 ABC patients with pathological diagnosis were separated into two groups: S-1 with or without the 3rd generation chemotherapy drug (group S-1) and capecitabine with or without the 3rd generation chemotherapy drug (Group capecitabine). The efficacy, side effects and prognostic factors were compared between the two groups. Results: There were 70 patients in group S-1 and 84 patients in group capecitabine. The objective response rates (ORR) were 31.4% (22/70) in group S-1 and 28.6% (24/84) in group capecitabine. The disease control rates (DCR) were 74.3% (52/70) and 83.3% (70/84), respectively. There were no significant differences in DCR and ORR between two groups (P>0.05). The DCR of patients treated with capecitabine monotherapy was significantly higher than that of S-1 monotherapy [94.4%(17/18) and 64.0%(16/25), P=0.028]. The median PFS was 7.5 and 8.9 months for the patients in the group S-1 and group capecitabine, with no statistically significant difference (P=0.423). The 1-year survival rates of group S-1 and group capecitabine were 81.4% and 66.7%, respectively, with no significant differences(P=0.020). Univariate analysis showed that ER and/or PR status (P=0.004), T stage (P=0.041), and molecular typing (P=0.046) were associated with PFS. Multivariate analysis showed ER and/or PR status (P=0.034) was an independent prognostic factor related with PFS. The incidence of hemoglobin reduction was 14.3% (10/70) and 36.9% (31/84) in the group S-1 and group capecitabine, and the differences were statistically significant (P=0.002). There was no significant difference in the incidence of leukopenia, neutropenia, thrombocytopenia and hand-foot syndrome between the two groups(P>0.05). Conclusions S-1 and capecitabine are both effective for advanced breast cancer. Neither ORR nor DCR were significantly different between these two groups. The incidence of gastrointestinal reactions and thrombocytopenia of S-1 was slightly lower than that of capecitabine.

Objective To compare the dosemetry between helical tomotherapy (HT) and intensity-modulated radiotherapy (IMRT) for early-stage postoperative breast cancer and provide more valuable evidences to the clinical researches. Methods Clinical trails of dosimetric comparing between HT and IMRT for early-stage breast cancer were obtained from PubMed, Embase, Sciencedirect, CNKI, VIP and Wanfang databases, evaluated and analyzed with the Cochrane Collaboration's RevMan 5.2 software. Results 10 studies were included with a total of 135 patients. Compared to IMRT plans, HT plans provided a significantly better conformity index (P<0.000 1), mean (P<0.000 01) and maximal dose (P=0.003) of the planning target volume (PTV). HT plans had a lower heart maximal dose (P=0.005), V20 (P=0.05), V30 (P=0.003), and ipsilateral lung maximal dose (P=0.003), V20 (P=0.02), as while as had a higher contralateral breast V5 (P=0.01), mean (P=0.05) and maximal dose (P<0.000 01). There was no significantly difference between HT and IMRT plans for homogeneity index of PTV, heart V5, V10, mean dose, ipsilateral lung V5, V10, V30, mean dose, contralateral breast V10, contralateral lung mean and maximal dose (all P >0.05). Conclusion Compared to IMRT plans, HT plans have the dosimetry superiority for early-stage breast cancer with significantly better coverage and dose conformity while maintaining lower doses to high risk organs.