Attenuated Salmonella typhimurium VNP20009 is a widely used natural oncolytic bacterium, which has great application potential given its unique characteristics, including clinical safety, tumor targeting specificity, and explicit genome sequence. Here, we show that tumor progression can be effectively reduced by intraperitoneal administration with VNP20009 in a mouse model of melanoma (all animal experiments were conducted in accordance with the Animal Ethics Committee of China Pharmaceutical University); co-culture experiment in vitro demonstrated that VNP20009 can induce the polarization of macrophage M1, accompanied by expression of inflammation-related factors; flow cytometry analysis showed that VNP20009 induced the increase of immune cell infiltration in tumor. Further analysis showed that T cells infiltration in tumor-draining lymph node (TDLN) increased, and VNP20009 induced the activation of CD4⁺ T cells and CD8⁺ T cells in tumor. Our results demonstrate that VNP20009 treatment significantly inhibited melanoma tumors by remodeling tumor-associated macrophages to an M1-like phenotype, as well as recruiting and activating cytotoxic T cells, combined with its own antigenic activity to exert anti-tumor immunity.

Objective:To construct radiomics models of micro-calcification in carotid plaques, and compare their diagnostic values.Methods:Fifty-two patients with large atherosclerotic cerebral infarction admitted to Department of Neurology, Third Affiliated Hospital of Soochow University from May 2017 to November 2019 were enrolled. All patients underwent conventional carotid artery Doppler ultrasound to detect carotid plaques and Micropure? ultrasound to detect micro-calcifications in the plaques. A cross-section image with maximum numbers of micro-calcifications was chosen when there were micro-calcifications in carotid plaques; otherwise, a cross-section image with the largest area of the plaque was chosen. After all images were normalized by Photoshop software, the plaques were delineated as regions of interest using MaZda 4.6 software and 283 texture features of the plaques were automatically extracted. The texture features with the strongest predictive value were selected through consistency analysis (intrclass correlation coefficient [ICC]>0.75), two-sample t-test, Least absolute shrinkage and selection operator (Lasso) regression. The predictive models were constructed by RandomForest (RF) and Support vector machine (SVM) classifiers. The training set and test set were divided by 7: 3 to analyze the classification accuracy. Receiver operating characteristic (ROC) curves were used to calculate the area under the curve (AUC) to evaluate the diagnostic values of the models. Delong test was used to compare the difference between the diagnostic values of the 2 classifiers in test set. Results:A total of 148 plaque images from 52 patients were enrolled, including 104 plaques with micro-calcification and 44 plaques without micro-calcification. Nine texture features were finally selected after ICC analysis, T test and Lasso regression: 5 image gray histogram features were mean, variance, skewness, kurtosis and 99 th percentile (Perc. 99%); 1 autoregressive model feature was Teta3, and 3 wavelet transform features were WavEnLH_s-3, WavEnLH_s-4, and WavEnLH_s-6. With RF classifier, accuracy of the diagnostic model was 0.93, enjoying AUC of 0.92; with SVM classifier, that was 0.91, enjoying AUC of 0.90; Delong test showed that the diagnostic values of the 2 classifiers in test set were significantly different ( Z=1.000, P=0.320). Conclusion:Radiomic models constructed by RF and SVM classifiers can identify micro-calcification in carotid plaques, and the 2 classifiers share equivalent diagnostic values.

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases

Purpose: Capecitabine is an extensively used oral prodrug of 5-fluorouracil in treatment of colon cancer and is known to cause hand-foot syndrome (HFS). As the target enzyme for capecitabine, thymidylate synthase (TYMS) plays a key role for 5-fluorouracil metabolism and has been associated with some side effects caused by capecitabine. The aim of our study is to identify the possible genetic predictors of capecitabine-induced HFS (CAP-HFS) in Chinese colorectal cancer patients. Materials and Methods: Whole exons of TYMS were sequenced for 288 extreme phenotype HFS patients, including 144 severe or early-onset (first 2 cycles) moderate HFS extreme cases and 144 extreme controls with no reported HFS. The associations between polymorphisms and CAP-HFS were analyzed using logistic regression under an additive model. Results: We identified a novel risk mutation (c.1A>G, chr18:657743), was associated with severe HFS in an extreme case who was affected during the first cycle of treatment. Moreover, we identified three new variants, rs3786362, rs699517, rs2790, and two previously reported variants, 5’VNTR 2R/3R and 3′-untranslated region 6-bp ins-del, which were significantly associated with CAP-HFS (p < 0.05). In silico analysis revealed that the effect of these polymorphisms in the TYMS region on the development of HFS might not be restricted solely to the regulation of TYMS expression, but also the TYMS catalytic activity through the indirect effect on ENOSF1 expression. Conclusion This study identified new polymorphisms in TYMS gene significantly associated with CAP-HFS, which may serve as useful genetic predictors for CAP-HFS and help to elucidate the underlying mechanism of HFS.

Purpose: Capecitabine is an extensively used oral prodrug of 5-fluorouracil in treatment of colon cancer and is known to cause hand-foot syndrome (HFS). As the target enzyme for capecitabine, thymidylate synthase (TYMS) plays a key role for 5-fluorouracil metabolism and has been associated with some side effects caused by capecitabine. The aim of our study is to identify the possible genetic predictors of capecitabine-induced HFS (CAP-HFS) in Chinese colorectal cancer patients. Materials and Methods: Whole exons of TYMS were sequenced for 288 extreme phenotype HFS patients, including 144 severe or early-onset (first 2 cycles) moderate HFS extreme cases and 144 extreme controls with no reported HFS. The associations between polymorphisms and CAP-HFS were analyzed using logistic regression under an additive model. Results: We identified a novel risk mutation (c.1A>G, chr18:657743), was associated with severe HFS in an extreme case who was affected during the first cycle of treatment. Moreover, we identified three new variants, rs3786362, rs699517, rs2790, and two previously reported variants, 5’VNTR 2R/3R and 3′-untranslated region 6-bp ins-del, which were significantly associated with CAP-HFS (p < 0.05). In silico analysis revealed that the effect of these polymorphisms in the TYMS region on the development of HFS might not be restricted solely to the regulation of TYMS expression, but also the TYMS catalytic activity through the indirect effect on ENOSF1 expression. Conclusion This study identified new polymorphisms in TYMS gene significantly associated with CAP-HFS, which may serve as useful genetic predictors for CAP-HFS and help to elucidate the underlying mechanism of HFS.

BACKGROUND: Fetal bovine serum is widely used as a growth supplement for cell culture medium; however, animalbornepathogens increase the risk of transmitting infectious agents. Platelet-rich fibrin is recently considered as a successfulalternative but leukocytes present limits to its allogeneic feasibility. The aim of this study was to explore the effects ofallogeneic fibrin clot (AFC) without leukocytes on inducing odontogenic/cementogenic differentiation of human dentalpulp stem cells (hDPSCs) and human periodontal ligament stem cells (hPDLSCs) in vitro and in vivo. METHODS: AFC was prepared by high-speed centrifugation and leukocytes were almost removed, and AFC serum wasobtained through three freeze–thaw cycles. hDPSCs and hPDLSCs were treated with AFC serum to investigate theodontogenic or cementogenic associated markers by real-time polymerase chain reaction. hDPSCs were treated with AFCserum and placed inside of dentin canal, hPDLSCs were treated with AFC serum to wrap outside of dentin, the mixture wasthen transplanted into the subcutaneous of nude mice for 12 weeks. RESULTS: AFC serum exhibited enough growth factors and cytokines to induce odontogenic/cementogenic differentiationof hDPSCs and hPDLSCs in vitro. Furthermore, AFC seurum could induce hDPSCs to differentiate into odontoblastslikecells and pulp-like tissues, and hPDLSCs to regenerate cementum-like tissues. CONCLUSION AFC could be an alternative safe source with growth factors for the expansion of human dental mesenchymalstem cells (hDMSCs).

BACKGROUND: Fetal bovine serum is widely used as a growth supplement for cell culture medium; however, animalbornepathogens increase the risk of transmitting infectious agents. Platelet-rich fibrin is recently considered as a successfulalternative but leukocytes present limits to its allogeneic feasibility. The aim of this study was to explore the effects ofallogeneic fibrin clot (AFC) without leukocytes on inducing odontogenic/cementogenic differentiation of human dentalpulp stem cells (hDPSCs) and human periodontal ligament stem cells (hPDLSCs) in vitro and in vivo. METHODS: AFC was prepared by high-speed centrifugation and leukocytes were almost removed, and AFC serum wasobtained through three freeze–thaw cycles. hDPSCs and hPDLSCs were treated with AFC serum to investigate theodontogenic or cementogenic associated markers by real-time polymerase chain reaction. hDPSCs were treated with AFCserum and placed inside of dentin canal, hPDLSCs were treated with AFC serum to wrap outside of dentin, the mixture wasthen transplanted into the subcutaneous of nude mice for 12 weeks. RESULTS: AFC serum exhibited enough growth factors and cytokines to induce odontogenic/cementogenic differentiationof hDPSCs and hPDLSCs in vitro. Furthermore, AFC seurum could induce hDPSCs to differentiate into odontoblastslikecells and pulp-like tissues, and hPDLSCs to regenerate cementum-like tissues. CONCLUSION AFC could be an alternative safe source with growth factors for the expansion of human dental mesenchymalstem cells (hDMSCs).

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires

Background: To evaluate the facial asymmetry, three-dimensional computed tomography (3D-CT) has been used widely. This study proposed a method to quantify facial asymmetry based on 3D-CT. Methods: The normal standard group consisted of twenty-five male subjects who had a balanced face and normal occlusion. Five anatomical landmarks were selected as reference points and ten anatomical landmarks were selected as measurement points to evaluate facial asymmetry. The formula of facial asymmetry index was designed by using the distances between the landmarks. The index value on a specific landmark indicated zero when the landmarks were located on the three-dimensional symmetric position. As the asymmetry of landmarks increased, the value of facial asymmetry index increased. For ten anatomical landmarks, the mean value of facial asymmetry index on each landmark was obtained in the normal standard group. Facial asymmetry index was applied to the patients who had undergone orthognathic surgery. Preoperative facial asymmetry and postoperative improvement were evaluated. Results: The reference facial asymmetry index on each landmark in the normal standard group was from 1.77 to 3.38. A polygonal chart was drawn to visualize the degree of asymmetry. In three patients who had undergone orthognathic surgery, it was checked that the method of facial asymmetry index showed the preoperative facial asymmetry and the postoperative improvement well. Conclusions The current new facial asymmetry index could efficiently quantify the degree of facial asymmetry from 3D-CT. This method could be used as an evaluation standard for facial asymmetry analysis.

OBJECTIVE To investigate the in vivo antitumor effect of S.pulvinata extract (SP) and its underlying mechanisms.METHODS A solid tumor H22-transplanted model in mice was established and randomly divided into normal saline control group,5-FU chemotherapy group (30 mg/kg),low-dose SP group (63 mg/kg),and high-dose SP group (189 mg/kg).All mice received respective treatment once daily for 15 days.The tumor weights were monitored,and the inhibition rates as well as organ indexes were calculated.The potential mechanism was investigated using TUNEL staining,qPCR and Western blot analysis.RESULTS Compared with the control group,SP significantly inhibited tumor growth without producing obvious side-effects on body weight and immune organs.Observations from a TUNEL staining experiment demonstrated that SP noticeably induced apoptosis in tumor tissues.Moreover,qPCR showed that SP upregulated the mRNA levels of Bax and downregulated those of Bcl-2,resulting in the release of cytochrome c,which is in agreement with their protein expressions.Further Western blot analysis revealed that SP significantly activated and induced the cleavage of caspase-3,caspase-8 and caspase-9.CONCLUSION The findings clearly demonstrates that the mechanism of SP involves the induction of apoptosis.The study suggests that SP possesses potent in vivo antitumor effects against HCC.