Enhancement of vitamin A combined vitamin D supplementation on immune response to Bacille Calmette-Guerin vaccine revaccinated in Chinese infants

Objective:To investigate whether there is an association between diameter of bacille CalmetteGuerin(BCG)sears and effect of purified protein derivative(PPD)reaction anil to determine whether vitamin A(VA)combined vitamin I)(VD)supplementation influences the immune response to BCG revueeinated in Chinese infants.Methods:A cross-section and 3-month community-randomised trial was conducted.A total of 5 629 infants at 3,6 and 12 months of age in Junan County of China were examined for BCG scar fonnation.Then,597 revuccinated infants were randomly assigned to supplementation(n=307)and control(n=290)groups.The supplementation group were daily assigned to 1 500 IU VA and 500 IU VD for 3 months.Then all infants were subjected to skin test with PPD.Results:The diameter of BCG sears was positively con-elated with diameter of skin indurations of PPD(r=0.17,P<0.05)in the 5 629 infants.The rate of positive response to PP1)was higher in the supplementation group than in the control group(96.1%versus 89.7%,P<0.05,prevalence ratio 1.07.95%CI 1.02-1.12).The prevalence ralio of PPD response for the supplementation group compared with that for the control group was 1.07(95%CI 1.01-1.13)for the males and 1.08(95%CI 1.00-1.17)for the females.For the supplementation group,the males got larger tuberculin induration than the females[(0.73±0.2l)cm versus(0.67±0.20)cm.P<0.05)after intervention.Conclusions:The diameter of BCG scars was effectively correlated with PPD response,which indicates BCG scar formation may be an useful tool Io evaluate the effect of tuberculosis prevention.VA combined VD supplementation may play an immunoregulatory rale in BCG revuecination.This may contribute to the prevention of childhood tuberculosis.

The natural course of bacillus Calmette-Guerin induced bladder lesions:A long-term follow-up study and systematic review

Objective:Bacillus Calmette-Gue´rin(BCG)instillation is the standard adjuvant treatment for intermediate-and high-risk non-muscle-invasive bladder cancer after transurethral resection.Nevertheless,its toxicity often causes bladder complications.On follow-up cystoscopy,post-BCG bladder lesions can be pathologically benign,urothelial carcinoma recurrence,or other types of bladder malignancy.Only a small number of case reports have been published on post-BCG bladder lesions.Their clinical features,natural course,and management remain unknown.Methods:We retrospectively studied cystoscopic videos and medical records of BCG-treated bladder cancer patients at our center.During a long-term follow-up,we took biopsies on tumor-like lesions and described their changes.In addition,we summarized previous studies on post-BCG bladder lesions by systematic literature searching and review.Results:We described a series of three cases with post-BCG bladder lesions mimicking tumor recurrence from a total of 38 cases with follow-up data for more than 5 years.Those lesions could last,grow,or disappear spontaneously,and remain pathological benign for years.In systematic review,we identified and analyzed a total of 15 cases with post-BCG bladder lesions with detailed clinical information.Eleven of the 15 were benign and have a good prognosis with nephrogenic adenoma being the most common pathological type.Conclusion:Based on previous studies and our experience,benign lesions after BCG instillation cannot distinguish with cancer recurrence by cystoscopy alone,even under narrow band imaging mode.Nonetheless,given most of them have a good prognosis,random biopsy or transurethral resection might be spared in the patients with long-term negative biopsy and urine cytology.

Granulomatous prostatitis after bacille Calmette-Guérin instillation resembles prostate carcinoma:A case report and review of the literature

BACKGROUND Bacille Calmette-Guérin(BCG)instillation is recommended in patients with nonmuscle-invasive bladder cancer who have intermediate-risk and high-risk tumors.However,granulomatous prostatitis is a rare complication induced by BCG instillation,which can easily be misdiagnosed as prostate cancer.Here,we report a case of granulomatous prostatitis that resembled prostate cancer.CASE SUMMARY A 64-year-old Chinese man with bladder cancer received BCG instillation.Three days later,he stopped BCG instillation and received anti-infective therapy due to the urinary tract infection.Three months after BCG restart,he had rising total prostate-specific antigen(PSA)(9.14 ng/mL)and decreasing free PSA/total PSA(0.09).T2-weighted images of magnetic resonance imaging(MRI)showed a 28 mm×20 mm diffuse low signal abnormality in the right peripheral zone,which was markedly hyperintense on high b-value diffusion-weighted MRI and hypointense on apparent diffusion coefficient map images.Considering Prostate Imaging Reporting and Data System score of 5 and possibility of prostate cancer,a prostate biopsy was conducted.Histopathology showed typical features of granulomatous prostatitis.The nucleic acid test for tuberculosis was positive.He was finally diagnosed with BCG-induced granulomatous prostatitis.Thereafter,he stopped BCG instillation and received anti-tuberculosis treatment.During 10 mo follow-up,he had no evidence of tumor recurrence or symptoms of tuberculosis.CONCLUSION Temporarily elevated PSA and high followed by low signal abnormality on diffusion-weighted MRI are important indicators of BCG-induced granulomatous prostatitis.

Intravesical bacillus Calmette-Guerin(BCG)in treating non-muscle invasive bladder cancer—analysis of adverse effects and effectiveness of two strains of BCG(Danish 1331 and Moscow-I)

Objective:To compare the differences in adverse effects and efficacy profile between bacillus Calmette-Guerin(BCG)Danish 1331 and BCG Moscow-I strain in management of non-muscle invasive bladder cancer.Methods:Clinical data of 188 cases of non-muscle invasive bladder cancer treated with BCG between January 2008 and December 2018 in our institute were collected prospectively and analysed retrospectively,and 114 patients who completed a minimum of 12 months of follow-up were analysed.Patient and tumor characteristics,strain of BCG,adverse effects,and tumor progression were included for analysis.Intravesical BCG was instilled in intermediate-and high-risk patients.Six weeks of induction BCG,followed by three weekly maintenance BCG at 3,6,12,18,and 24 months was advised in high-risk patients.Results:Overall 68 patients received BCG Danish 1331 strain and 46 patients received Moscow-I strain.Patient and tumor characteristics were well balanced between the two groups.The median follow-up period was 42.5 months and 34.5 months in Danish 1331 and Moscow-I groups,respectively.Adverse events like dropout rate,antitubercular treatment requirement,and need of cystectomy were higher in Moscow-I group(n=31,67.4%)when compared to Danish 1331 strain(n=33,48.5%)(p=0.046).On direct comparison between Danish 1331 and Moscow-I strain,there was similar 3-year recurrence-free survival(80.0%vs.72.9%)and 3-year progression-free survival(96.5%vs.97.8%).Conclusion:Study results suggest no significant differences between Danish 1331 and Moscow-I strain in recurrence-free survival and progression-free survival,but a significantly higher incidence of moderate to severe adverse events in BCG Moscow-I strain.

Repeated inoculations of Mycobacterium bovis Bacille Calmette-Guérin (BCG) are needed to induce a strong humoral immune response against antigens expressed by the bacteria

The cellular immune response elicited by Mycobacterium bovis Bacille Calmette-Guérin (BCG) has been carefully investigated, but the humoral immune response has been partially neglected. BALB/c mice were immunized with BCG strain used to immunize humans. Anti-BCG antibodies, as assayed by ELISA, began to appear in the sera after the third week of immunization and plateaued three weeks after the 8th immunization. The total immunoglobulins (Igs) were purified by caprylic acid method from pooled serum collected after the 8th immunization. Anti-BCG antigen antibodies were detected in the total Igs preparation as well as in IgG, IgM, IgA, IgG1, IgG2a, and IgG2b, but not in the IgG3. Distinct BCG proteins were recognized the IgGs in Western blot analysis. Opsonization of BCG bacilli by the purified Igs potentiated internalization of the bacteria by murine Raw 264.7 macrophages. The intracellular BCG elimination coincided with the induction of NO production, which was more pronounced in cells infected with opsonized BCG compared to those infected with the non-opsonized bacteria. Coincidently, the production of NO was also higher in macrophages infected with opsonized BCG (maximal NO production at 48 h of incubation). The obtained results demonstrate that repeated inoculations of BCG effectively activate the humoral immune response, justifying the use of BCG as a live recombinant vaccine vector to insert genes encoding virulence factors controlled by antibodies.

Multi-organ failure with atypical liver granulomas following intravesical Bacillus Calmette-Guerin instillation

Bacillus Calmette-Guerin(BCG) intravesical instillation has been adopted in the treatment of patients with superficial bladder cancer.BCG-induced disseminated infection,though rare,has been associated with the histological finding of epithelioid granulomas in different organs,including the liver.We report the case of an adult patient with multi-organ failure,who developed sepsis,acute respiratory failure and acute hepatic failure with encephalopathy whose liver biopsy confirmed the presence of atypical,granulomatous-like lesions.Recovery was observed only after empirical therapy for Mycobacterium bovis with isoniazid,rifampicin,ethambutol and steroids was introduced.This case highlights the importance of a thorough patient assessment in order to exclude other more common causes of hepatic granulomas and to confirm diagnosis.Histological findings may be non-specific when the liver is involved in BCGinduced disseminated infection.

Effects of Bacillus amyloliquefaciens Biofertilizer on Brassica juncea var. multiceps Growth and N_2O Emission from Soil

In this study, Bacil us amyloliquefaciens A3 was continual y incubated in shake fIasks contalning wastewater from sweet potato starch production as an ef-fective biofertiIizer for cuItivation of Brassica juncea var. multiceps（XueIihong）. Based on pot experiments in the greenhouse, the effects of chemical fertiIizers （CN）, biofertiIizer （BF）, inactivated broth （BI）, starch wastewater （SW） and the combination of biofertiIizer and chemical fertiIizer （BC） on the yield, NO3- content and NO2- con-tent of XueIihong, soiI physicochemical properties and N2O emission were investi-gated. The resuIts showed that the yield of XueIihong in BC and CN treatments was improved by five times compared with CK; BF and SW treatments had insignifi-cant impact on the yield of XueIihong. Compared with CN treatment, BCL treatment exhibited simiIar improving effects on the yield of XueIihong, in which NO3- content of XueIihong and soiI was reduced by 16.4%-73.6% and 22%-29%, which reduced the risk of nitrogen eIuviations in soiI; average N2O fIux （FPV30） in BCL treatment was reduced by 58.3%-73.1% compared with CN treatment. In concIusion, B. amy-loliquefaciens is a feasibIe Iow-cost biofertiIizer for sustalnabIe vegetabIe farming with a great potential for starch wastewater utiIization.

Control Effects of Bacillus subtilis DJ-6 and Pyraclostrobin Alone and in Combination Against Fusarium oxysporum

[Objective] This study was conducted to screen a synergistic biological fungicide complex to control Fusarium wilt, reducing the use of chemical pesticides. [Method] The inhibitory effects of Bacil us subtilis DJ-6 and pyraclostrobin alone or in combination at five ratios against Fusarium oxysporum were detected by mea-suring mycelium growth rate in laboratory tests. The growth promotion and disease control effect of combined or single use of 20% pyraclostrobin and 2 ×1011 cfu/g B. subtilis DJ-6 WP at 1∶1 000, 1∶2 000 and 1∶3 000 dilutions were detected in field trials. [Result] The EC50 values of combined use of B. subtilis DJ-6 and pyra-clostrobin at ratios of 1∶1, 1∶2, 1∶3, 1∶4 and 1∶5 against F. oxysporum were 5.311 5, 4.008 6, 3.570 6, 3.350 9 and 3.218 9 μg/ml, with the synergistic ratios （SR） of 2.28, 1.77, 1.53, 1.64, 1.11, among which the synergetic effect at 1∶1 was the best. The fungicidal activity of pyraclostrobin was greater than that of B. subtilis DJ-6 in laboratory tests. Field trials revealed that al the 1∶1 000, 1∶2 000 and 1∶3 000 dilu-tions of 20% pyraclostrobin·2×1011 cfu/g B. subtilis DJ-6 WP in combination, 1∶1 000 dilution of 1 ×1012 cfu/g B. subtilis DJ-6 WP and 1∶2 000 dilution of 250 g/L pyra-clostrobin EC promoted the growth of strawberry by increasing plant height, leaf petiole, leaf blade area and stem diameter. Among them, the treatments with 1∶1 000 and 1∶2 000 of 20% pyraclostrobin · 2×1011 cfu/g B. subtilis DJ-6 WP in combina-tion had better effects than other treatments. The control effects of al the treat-ments were measured 30 and 80 d after fungicide application. The control effects of 1∶1 000 dilution of 20% pyraclostrobin and 2×1011 cfu/g B. subtilis DJ-6 in combina-tion were up to 100% and 93.11%, which were higher than those in al other treat-ments. The second highest control effects were found in the treatment with 1∶ 2 000 dilution of 20% pyraclostrobin and 2×1011 cfu/g B. subtilis DJ-6 in combination, they were 92.49% and 86.49%, higher than those in other treatments except the 1∶1 000 dilution of 20% pyraclostrobin and 2 ×1011 cfu/g B. subtilis DJ-6 in combination. The control effects of 1∶3 000 dilution of 20% pyraclostrobin and 2×1011 cfu/g B. subtilis DJ-6 in combination were 82.61% and 72.42%, higher than those in treatment with 1∶1 000 dilution of 1×1012 cfu/g B. subtilis DJ-6 WP, but lower than those in treat-ment with 1∶2 000 dilution of 25% pyraclostrobin EC. [Conclusion] Al the results re-vealed that the combination use of 20% pyraclostrobin and 2 ×1011 cfu/g B. subtilis DJ-6 WP at 1∶1 000 to 1∶2 000 dilution had better control effect against strawberry Fusarium wilt.

Effects of Bacillus amyloliquefaciens Biofertilizer on Tea Yield and Quality

[Objective] This study was aimed to explore the effects of Bacil us amy-loliquefaciens biofertilizer on tea yield and quality. [Method] The field plot experiment was conducted with the biofertilizer treatments and control to investigate 100-bud weight and main biochemical components. [Result] The treatments by Bacil us amy-loliquefaciens biofertilizer, which was fermented using sweet potato starch wastewa-ter （SPSW） as culture medium, improved 100-bud weight and tea quality significant-ly under the concentration of 0.8×108, 1.6×108 and 3.2×108 cfu/ml with the dose of 1 L/m2 for 4 times. At the optimum concentration of 1.6 ×108 cfu/ml, the biofertilizer treatment increased the 100-bud weight by 22.3%, water extracting materials by 21.9%, amino acids content by 8.83%, tea polyphenol content by 9.76%, and de-creased theine content by 8.32%, respectively. Compared with the control, there was no significant difference between the SPSW treatment and the control. [Con-clusion] The production of the B. amyloliquefaciens biofertilizer could consume SP-SW, and the application of the biofertilizer could improved the tea yield and quality, which provided references for the development of ecological agriculture.

Bacillus sp. probiotic supplementation diminish the Escherichia coli F4ac infection in susceptible weaned pigs by influencing the intestinal immune response, intestinal microbiota and blood metabolomics

Background:Probiosis is considered a potential strategy to reduce antibiotics use and prevent postweaning diarrhea(PWD).This study investigated the effect of Bacillus amyloliquefaciens DSM25840 or Bacillus subtilis DSM25841 supplementation on growth,health,immunity,intestinal functionality and microbial profile of post-weaning pigs after enterotoxigenic E.coli(ETEC)F4 challenge.Methods:Sixty-four post-weaning piglets(7748 g±643 g)were randomly al ocated to four groups:control basal diet(CO);CO+1.28×10~6 CFU/g of B.amyloliquefaciens(BAA);CO+1.28×10~6 CFU/g feed of B.subtilis(BAS);CO+1 g colistin/kg of feed(AB).At day(d)7,animals were chal enged with 10~5 CFU/m L of ETEC F4 ac O149 and then fol owed for fecal score and performance until d 21.Blood was col ected at d 6,d 12 and d 21 for immunoglobulins,at d 8 for acute phase proteins,at d 8 and d 21 for metabolomics analysis.Jejunum was sampled for morphometry,quantification of apoptosis,cel proliferation,neutral and acid mucine and Ig A secretory cel s,and microarray analysis at d 21.Jejunum and cecum contents were col ected for microbiota at d 21.Results:AB and BAS reduced the fecal score impairment compared to CO(P<0.05)at d 14.Body weight(BW),average daily weight gain(ADWG),average daily feed intake(ADFI)and gain to feed ratio(G:F)did not differ between Bacil us groups and CO.AB improved BW at d 7,d 14 and d 21,ADWG ADFI and G:F from d 0 to d 7(P<0.05).At d 8,CO had higher plasma arginine,lysine,ornithine,glycine,serine and threonine than other groups,and higher haptoglobin than AB(P<0.05).At d 21,CO had lower blood glycine,glutamine and Ig A than BAS.Morphology,cel s apoptosis and mucins did not differ.BAS and AB increased the vil us mitotic index.Transcriptome profile of BAS and AB were more similar than CO.Gene sets related to adaptive immune response were enriched in BAA,BAS and AB.CO had enriched gene set for nuclear structure and RNA processing.CO had a trend of higher Enterobacteriaceae in cecum than the other groups(P=0.06).Conclusion:Bacil us subtilis DSM25841 treatment may reduce ETEC F4ac infection in weaned piglets,decreasing diarrhea and influencing mucosal transcriptomic profile.

Effects of Bacillus coagulans supplementation on the growth performance and gut health of broiler chickens with Clostridium perfringens-induced necrotic enteritis

Background: The poultry industry is in need of effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE) due to Clostridium perfringens.Methods: This study was conducted to investigate the effects of feeding Bacil us coagulans on the growth performance and gut health of broiler chickens with C. perfringens-induced NE. Two hundred and forty 1-day-old broiler chicks were randomly assigned to a 2 × 2 factorial arrangement with two dietary B. coagulans levels(0 or 4 × 109 CFU/kg of diet) and two disease chal enge statuses(control or NE chal enged).Results: NE-induced reduction in body weight gain was relieved by the addition of B. coagulans into broiler diets compared with the NE-infected birds. NE infection damaged intestinal morphological structure, promoted intestinal C.perfringens growth and liver invasion, and enhanced anti-C. perfringens specific sI gA concentrations in the gut and specific IgG levels in serum compared with the uninfected birds. NE infection significantly(P < 0.05) decreased mucin-2(at 14 d post-infection(DPI), tol-like receptor 2(TLR2, at 7 and 14 DPI), TLR4(at 7 and 14 DPI), tumor necrosis factor super family15(TNFSF15, at 7 and 14 DPI), lysozyme(LYZ, at 14 DPI) and fowlicidin-2(at 7 and 14 DPI) mR NA levels, whereas it dramatical y(P = 0.001) increased IFN-γ mR NA levels at 7 DPI. However, chal enged birds fed diets supplemented with B.coagulans showed a significant(P < 0.01) decrease in gut lesion scores, decreased C. perfringens numbers in the cecum and liver, and an increase in fowlicidin-2 mR NA levels in compared with the uninfected birds. In addition, compared with the non-supplemented group, dietary inclusion of B. coagulans improved intestinal barrier structure, further increased specific sI gA levels and alkaline phosphatase(IAP) activity in the jejunum, enhanced the expression of jejunum lysozyme mR NA, and inhibited the growth, colonization, and invasion of C. perfringens; in contrast, it reduced serum-specific IgG concentrations and jejunum IFN-γ mR NA levels.Conclusion: These results indicated that dietary B. coagulans supplementation appeared to be effective in preventing the occurrence and reducing the severity of C. perfringens-induced NE in broiler chickens.

Characterization of two alkyl hydroperoxide reductase C homologs alkyl hydroperoxide reductase C_H1 and alkyl hydroperoxide reductase C_H2 in Bacillus subtilis

AIM: To identify alkyl hydroperoxide reductase subunit C(AhpC) homologs in Bacillus subtilis(B. subtilis) and to characterize their structural and biochemical properties. AhpC is responsible for the detoxification of reactive oxygen species in bacteria.METHODS: Two AhpC homologs(AhpC_H1 and AhpC_H2) were identified by searching the B. subtilis database; these were then cloned and expressed in Escherichia coli. AhpC mutants carrying substitutions of catalytically important Cys residues(C37S, C47 S, C166 S, C37/47 S, C37/166 S, C47/166 S, and C37/47/166 S for AhpC_H1; C52 S, C169 S, and C52/169 S for AhpC_H2) were obtained by site-directed mutagenesis and purified, and their structure-function relationship was analyzed. The B. subtilis ahp C genes were disrupted by the short flanking homology method, and the phenotypes of the resulting AhpC-deficient bacteria were examined.RESULTS: Comparative characterization of AhpC homologs indicates that AhpC_H1 contains an extra C37, which forms a disulfide bond with the peroxidatic C47, and behaves like an atypical 2-Cys AhpC, while AhpC_H2 functions like a typical 2-Cys AhpC. Tryptic digestion analysis demonstrated the presence of intramolecular Cys37-Cys47 linkage, which could be reduced by thioredoxin, resulting in the association of the dimer into higher-molecular-mass complexes. Peroxidase activity analysis of Cys→Ser mutants indicated that three Cys residues were involved in the catalysis. AhpC_H1 was resistant to inactivation by peroxide substrates, but had lower activity at physiological H2O2 concentrations compared to AhpC_H2, suggesting that in B. subtilis, the enzymes may be physiologically functional at different substrate concentrations. The exposure to organic peroxides induced AhpC_H1 expression, while AhpC_H1-deficient mutants exhibited growth retardation in the stationary phase, suggesting the role of AhpC_H1 as an antioxidant scavenger of lipid hydroperoxides and a stress-response factor in B. subtilis. CONCLUSION: AhpC_H1, a novel atypical 2-Cys AhpC, is functionally distinct from AhpC_H2, a typical 2-Cys AhpC.

Effects of dietary Bacillus amyloliquefaciens supplementation on growth performance,intestinal morphology, inflammatory response, and microbiota of intra-uterine growth retarded weanling piglets

Background: The focus of recent research has been directed toward the probiotic potential of Bacillus amyloliquefaciens(BA) on the gut health of animals. However, little is known about BA's effects on piglets with intra-uterine growth retardation(IUGR). Therefore, this study investigated the effects of BA supplementation on the growth performance,intestinal morphology, inflammatory response, and microbiota of IUGR piglets.Methods: Eighteen litters of newborn piglets were selected at birth, with one normal birth weight(NBW) and two IUGR piglets in each litter(i.e., 18 NBW and 36 IUGR piglets in total). At weaning, the NBW piglet and one of the IUGR piglets were assigned to groups fed a control diet(i.e., the NBW-CON and IUGR-CON groups). The other IUGR piglet was assigned to a group fed the control diet supplemented with 2.0 g BA per kg of diet(i.e., IUGR-BA group). The piglets were thus distributed across three groups for a four-week period.Results: IUGR reduced the growth performance of the IUGR-CON piglets compared with the NBW-CON piglets. It was also associated with decreased vil us sizes, increased apoptosis rates, reduced goblet cel numbers, and an imbalance between pro-and anti-inflammatory cytokines in the smal intestine. Supplementation with BA improved the average daily weight gain and the feed efficiency of the IUGR-BA group compared with the IUGR-CON group(P < 0.05). The IUGR-BA group exhibited increases in the ratio of jejunal vil us height to crypt depth, in ileal vil us height, and in ileal goblet cel density. They also exhibited decreases in the numbers of jejunal and ileal apoptotic cel s and ileal proliferative cel s(P < 0.05). Supplementation with BA increased interleukin 10 content, but it decreased tumor necrosis factor alpha level in the smal intestines of the IUGR-BA piglets(P < 0.05). Furthermore, compared with the IUGR-CON piglets, the IUGR-BA piglets had less Escherichia coli in their jejunal digesta, but more Lactobacil us and Bifidobacterium in their ileal digesta(P < 0.05).Conclusions: Dietary supplementation with BA improves morphology, decreases inflammatory response, and regulates microbiota in the smal intestines of IUGR piglets, which may contribute to improved growth performance during early life.

卡介苗对1型糖尿病小鼠的免疫治疗作用及其机制

目的评价卡介苗(BCG)对1型糖尿病(T1DM)小鼠的免疫治疗效果,并探索其作用机制。方法将15只SPF级雄性C57BL/6小鼠随机分为造模组(n=10,腹腔注射50 mg/kg链尿佐菌素建立T1DM模型)与对照组(n=5,腹腔注射等量柠檬酸缓冲液);取造模成功即随机血糖≥16.7 mmol/L的10只小鼠分为T1DM组(n=5)与卡介苗(BCG)组(n=5)。随后BCG组小鼠经皮下注射BCG 1×10^(6) cfu/只进行免疫治疗,4周后加强免疫治疗1次,T1DM组与对照组注射相同剂量磷酸盐缓冲液(PBS)。在实验周期(13周)内,每周监测小鼠体重和进食量变化,尾静脉采血检测各组小鼠血糖水平,采用口服葡萄糖耐量实验(OGTT)检测各组小鼠葡萄糖负荷后的血糖调节能力。采用HE染色观察各组小鼠胰腺组织病理变化,免疫组化染色检测胰腺胰岛素水平,酶联免疫吸附法(ELISA)检测各组小鼠血清C-肽含量,实时荧光定量PCR(qRT-PCR)检测各组小鼠脾组织细胞因子mRNA水平,流式细胞术检测各组小鼠脾细胞中调节性T细胞(Treg)比例。结果BCG免疫治疗2次后(第13周),T1DM组小鼠血糖水平明显高于对照组(P<0.001),而BCG组血糖水平虽高于对照组(P<0.01),但明显低于T1DM组(P<0.01)。OGTT结果显示,3组小鼠在葡萄糖灌胃后血糖水平迅速升高,120 min时开始下降,到180 min时,对照组小鼠血糖水平降至正常水平;与对照组比较,T1DM组及BCG组小鼠在120 min后血糖水平虽有下降趋势,但在180 min时仍明显高于对照组(P<0.001),而BCG组与T1DM组间差异无统计学意义(P>0.05)。在初次免疫时(第5周),与对照组比较,T1DM组和BCG组小鼠体重增长百分比明显降低(P<0.0001),进食量明显增加(P<0.0001)。BCG免疫治疗2次后(第13周),与对照组比较,T1DM组小鼠体重增长百分比仍明显降低(P<0.0001),进食量明显增加(P<0.0001);与T1DM组比较,BCG组小鼠体重增长百分比增高(P<0.001),进食量减少(P<0.001),且与对照组比较差异无统计学意义(P>0.05)。HE染色结果显示,与对照组比较,T1DM组小鼠胰岛发生明显的萎缩,而BCG组小鼠胰岛形态与对照组接近,且胰岛内细胞数目明显多于T1DM组。免疫组化染色结果显示,与对照组比较,T1DM组小鼠胰腺中胰岛素阳性面积明显减少(P<0.01),而BCG组胰岛素阳性面积虽然少于对照组(P<0.05),但明显多于T1DM组(P<0.05)。ELISA检测结果显示,与对照组比较,T1DM组、BCG组血清C-肽含量明显降低(P<0.001,P<0.05),但BCG组血清C-肽含量明显高于T1DM组(P<0.05)。qRT-PCR结果显示,与对照组比较,T1DM组脾组织细胞因子mRNA水平无明显变化(P>0.05);BCG组白细胞介素-2(IL-2)、IL-10、转化生长因子-β(TGF-β)mRNA水平与对照组和T1DM组比较均明显升高(P<0.0001),而γ干扰素(IFN-γ)mRNA水平无明显变化(P>0.05)。流式细胞术检测结果显示,与对照组比较,T1DM组小鼠体内Treg细胞百分比降低(P<0.05);与T1DM组比较,BCG组小鼠Treg细胞百分比明显增高(P<0.05),且与对照组比较差异无统计学意义(P>0.05)。结论BCG免疫治疗可诱导Treg细胞增多,调节自身免疫应答,并通过减轻胰腺病理损伤、恢复胰岛细胞功能以促进胰岛素分泌,从而降低T1DM小鼠的血糖水平。

左氧氟沙星联合纤维素酶在体外能有效根除卡介苗生物被膜感染

目的 探究左氧氟沙星(LEV)和纤维素酶(CL)对结核杆菌模式菌—卡介苗(BCG)生物被膜的体外抗菌效果。方法 采用XTT法与结晶紫染色法确定BCG生物被膜的成熟生长周期。不同浓度的LEV和CL单独或联用处理BCG浮游菌及BCG生物被膜,采用结晶紫染色定量生物被膜中生物量的变化。然后将成熟的BCG生物被膜用CL单独处理24 h,通过SYTO 9染色、Calcofluor White Stain染色后,在共聚焦激光扫描显微镜观察活菌数量、生物被膜中纤维素含量变化,扫描电子显微镜下观察生物被膜的结构变化。结果 采用微量肉汤稀释法检测LEV的MIC、MBC和MBEC值分别为4、8和1024μg/mL。浓度为2.56、5.12和10.24 U/mL的CL与1/4×MIC抗生素(LEV)的组合会导致生物被膜生物量减少(P<0.001)。CL在10.24、5.12和2.56 U/mL3种浓度下对成熟BCG生物被膜均表现出明显的分散效应(P<0.001)。结论 当左氧氟沙星与纤维素酶联合使用时,抗生素能有效根除BCG生物被膜感染。

N-CWS和BCG膀胱灌注对非肌层浸润性膀胱癌术后疗效及生活质量评价:一项基于2年临床随访的回顾性研究

目的:探讨红色诺卡菌细胞壁骨架(nocardia rubra cell wall skeleton,N-CWS)和卡介苗(bacillus calmette-guerin,BCG)膀胱灌注对非肌层浸润性膀胱癌(non-muscle invasive bladder cancer,NMIBC)患者手术的疗效和生活质量的影响。方法:纳入2016年4月至2021年4月在四川省人民医院泌尿外科收治的86例中高危NMIBC患者,均行手术治疗,并具有完整的临床资料和随访数据。根据膀胱灌注药物,将患者分成N-CWS组(n=41)和BCG组(n=45)。随访2年比较2组患者生活质量、临床疗效及不良反应等指标。结果:N-CWS组患者6个月复发率为4.88%,低于BCG组的6.67%,差异无统计学意义(χ^(2)=0.125,P=0.723);N-CWS组患者1年复发率为9.76%,低于BCG组的17.78%,差异无统计学意义(χ^(2)=1.150,P=0.284);N-CWS组患者2年复发率为21.95%,低于BCG组的28.89%,差异无统计学意义(χ^(2)=3.268,P=0.071)。灌注后6个月N-CWS组患者角色功能、情绪功能及整体功能明显高于BCG组(P<0.05);N-CWS组患者疼痛、泌尿系统症状评分和治疗产生问题评分明显低于BCG组(P<0.05)。灌注后1年N-CWS组患者角色功能、情绪功能及整体功能明显高于BCG组(P<0.05);N-CWS组患者疼痛、泌尿系统症状评分和治疗产生问题评分明显低于BCG组(P<0.05);灌注后2年N-CWS组患者情绪功能高于BCG组(P<0.05),其余各项评分2组比较差异均无统计学意义。N-CWS组患者尿路刺激征、血尿、关节疼痛及发热等不良反应事件发生率明显低于BCG组(P<0.05)。结论:与BCG治疗相比,N-CWS应用于NMIBC术后膀胱灌注治疗疗效相当,可有效提高生活质量,且安全性高,临床可进一步研究并推广。

Effects of Bacillus Calmette-Guerin on immunometabolism,microbiome and liver diseases

Metabolic diseases have overtaken infectious diseases as the most serious public health issue and economic burden in most countries.Moreover,metabolic diseases increase the risk of having infectious diseases.The treatment of metabolic disease may require a long-term strategy of taking multiple medications,which can be costly and have side effects.Attempts to expand the therapeutic use of vaccination to prevent or treat metabolic diseases have attracted significant interest.A growing body of evidence indicates that Bacillus Calmette-Guerin(BCG)offers protection against non-infectious diseases.The non-specific effects of BCG occur likely due to the induction of trained immunity.In this regard,understanding how BCG influences the development of chronic metabolic health including liver diseases would be important.This review focuses on research on BCG,the constellation of disorders associated with metabolic health issues including liver diseases and diabetes as well as how BCG affects the gut microbiome,immunity,and metabolism.

狼毒类中药对结核杆菌抗菌作用的比较

对１２种狼毒类中药的乙醇提取物进行了结核杆菌的体外抑菌试验，实验结果表明，１２种狠毒均有不同程度的抑菌作用。其中以狠毒大戟（Ｅｕｐｈｏｒｂｉａｆｉｓｃｈｅｒｉａｎａ）根、大狠毒（Ｅ．ｎｅｍａｔｏｃｙｐｈａ）根的抑菌作用最强。

卡介苗多糖核酸与减毒活卡介苗治疗哮喘患儿疗效比较及治疗前后细胞因子的变化

目的探讨卡介苗多糖核酸(BCG-PSN)与减毒活卡介苗治疗儿童哮喘临床疗效及其对哮喘患儿TH1/TH2调节的差异。方法急性发作期哮喘患儿60例随机分A、B、C3组,每组20例。A组予全球哮喘防治创议(GI NA方案)+BCG-PSN0.5mg/次,肌注,1次/隔日,共36次;B组予GI NA方案+减毒活卡介苗0.1mL,皮内注射,仅1次;C组单纯GI NA方案治疗。酶联法检测治疗前后血清IgE及外周血单个核细胞(PBMC)诱生的IL-4、IL-5及IFN-γ。结果1.疗效:A组优于B组,B组优于C组(P均<0.05)。2.治疗组治疗后IFN-γ均高于治疗前,IL-4、IL-5、IgE均低于治疗前。对于调整IL-5,A组优于B组和C组(P<0.05,0.01);对于上调IFN-γ及下调IL-4、IgE,A与B组比较无显著差异(P>0.05)。结论1.BCG-PSN的疗效优于减毒活卡介苗。2.减毒活卡介苗一次性小剂量接种对哮喘儿IL-4、IFN-γ、IgE失衡的调节与BCG-PSN多次肌注效果相当,对IL-5失衡的调节不及BCG-PSN。