Attenuated Salmonella VNP20009 mutant (ΔhtrA) is a promising candidate for bacteria-mediated tumour therapy in hosts with TNFR1 deficiency.

VNP20009 is a genetically modified strain of Salmonella typhimurium and has a good anticancer effect wildly used in tumour therapy on animal models. For its clinical application, an accurate bio-safety assessment on sensitive models is necessary. In this study, we use TNFR1 KO mice as a susceptive model to assess the virulence of bacterial VNP20009 and its derivative ΔhtrA. By intraperitoneal administration of Salmonella, the increased lethality was observed in TNFR1 KO mice infected with VNP20009, but not with ΔhtrA. We performed a systemically comparative analysis of their toxicity, and ΔhtrA shows a better bio-safety for TNFR1 KO mice. Since the macrophages with TNFR1 deficiency exhibit a reduced ability of bacteria clearance, ΔhtrA with lower survival ability in normal macrophages restores its viability in TNFR1 KO macrophages. Thus, ΔhtrA was further tested for its antitumour effect in TNFR1 KO mice bearing a B16F10 melanoma model. It displays a moderate antitumour effect, suggesting ΔhtrA instead of VNP20009 might be a promising candidate for bacteria-mediated tumour therapy specific to those with low immunity. SIGNIFICANT AND IMPACT OF THE STUDY: VNP20009 is attenuated Salmonella with a good safety widely used for tumour-targeting bacterial therapies. Little is known about its toxicity in hosts with low immunity. This study is the first systemically comparative analysis of their toxicity of VNP20009 and its mutant ΔhtrA in TNFR1-KO mice. Research on toxicity of tumour-targeting Salmonella in mice with immunodeficiency can facilitate the optimization of bacterial therapies with reduced adverse effects in future clinical trials.

[Impact of psoas muscle index on early postoperative mortality and complications after liver transplantation].

Objective: To investigate the relationship between psoas muscle index (PMI) and early postoperative survival rate and the incidence of complications after liver transplantation in adults. Methods: The clinical data of 225 patients (male, n=184; female, n=41) underwent liver transplantation at the Organ Transplantation Department of First Central Clinic Institute of Tianjin Medical University from January 2014 to December 2016 were analyzed, retrospectively.Original disease: hepatitis B liver cirrhosis(44 cases), hepatitis C cirrhosis(10 cases), autoimmune liver cirrhosis(29 cases), other benign liver diseases(24 cases), liver cirrhosis with liver cancer(116 cases), hilar cholangiocarcinoma(1 case) and hepatic vascular sarcoma(1 case). The area of bilateral psoas muscle on the lower edge level of the third lumbar vertebral body was measured through preoperative CT image.The PMI was calculated using this formula: bilateral psoas muscle area (mm(2))/the square of the body height (m(2)). According to the receiver operating characteristic curve and cut-off values, the male and female patients were divided into low PMI group and high PMI group respectively.The χ(2) test, Fisher exact test and t-test was used to compare the differences in perioperative data, survival rate and postoperative complications between the two groups, respectively. Results: There were 44 patients in the low PMI group, and 181 patients in the high PMI group. ICU time was longer (82.5(62.0-128.0) hours vs.69.1(56.0-104.0) hours; P=0.006) and preoperative blood urea nitrogen level (5.86(4.35-15.52) mmol/L vs. 4.94(4.05-7.06) mmol/L; P=0.012) was higher in the low PMI group than those in the high PMI group. Incidence rates of grade 5 complication (18.2%) and grade 4a complication (18.2%) were higher in the low PMI group, and 120-day cumulative survival rate was lower than that in high PMI group(81.8% vs. 95.6%, P=0.001). On the other hand, there were no significant differences in preoperative white blood cell count level, serum creatinine level, operative time, anhepatic period time, intraoperative blood loss, and incidence of postoperative grade 3 complications between the two groups(all P>0.05). Conclusions: There is a significant correlation between PMI and early postoperative survival rate and incidence of complications.Patients with lower PMI has poor prognosis after liver transplantation.

The cytological effects of HBCDs on human hepatocyte L02 and the potential molecular mechanism.

The concentration of hexabromocyclododecanes (HBCDs) in the environment media and organism samples are gradually rising with the increase of HBCDs usage. This study is designed to investigate the cytological effects of HBCDs on human hepatocyte L02 and explore the potential molecular mechanism. The results of CCK-8 assay showed that high concentration of HBCDs (>20 µM) significantly suppressed cell survival, while comparatively lower dose of HBCDs (10(-13)-10(-7)M) slightly stimulated cell proliferation (P < 0.05). In the mean time, high concentration HBCDs markedly induced cell apoptosis and DNA damage, accompanying with increase of intracellular Ca(2+)level and decrease of mitochondrial membrane potential (P < 0.05). ROS level induced by low concentration of HBCDs was comparatively lower than that by high concentration of HBCDs. In addition, low concentration HBCDs exposure (10(-13)-10(-7)M) resulted in up-regulation of PCNA protein expression level in a time-dependent manner. However, high concentration HBCDs exposure led to increase of Apaf-1 expression level. In conclusion, loss of mitochondrial membrane potential and activation of Apaf-1 mediated pathway involve the L02 cell apoptosis induced by high concentration HBCDs exposure. However, low concentration HBCDs exposure could stimulate cell proliferation of L02 cells, which might be associated with enhancement of PCNA expression.

The psoas muscle depletion index is related to the degree of cirrhosis and skeletal muscle loss in patients with end-stage liver disease.

Objective: To establish a new psoas muscle depletion index (PDI) from healthy young donors and to explore the correlation between the PDI and the severity of cirrhosis in patients with endstage liver disease (ESLD). Methods: Clinical data of 461 healthy donors were collected during the period 2014-2019, and clinical data of 331 patients with ESLD were collected during the period 2014-2018. The patients were divided into four groups by PDI severity: PDI ≥ 0.90, PDI = 0.75-0.90, PDI = 0.50-0.75 and PDI ≤ 0.50 (Gsev). Differences in international normalised ratio (INR), total bilirubin and serum creatinine levels, and Child-Pugh (CP) and model for end-stage liver disease (MELD) scores were compared. The sarcopenia incidence according to the PDI and the psoas muscle index (PMI) in different weight groups were also compared. Results: Gsev had the highest CP (10.2 ± 2.1) and MELD (20.1 ± 7.4) scores and total bilirubin (166.3 ± 192.0 umol/L) and blood creatinine (92.9 ± 90.2 umol/L) levels and the lowest haemoglobin (93.8 ± 21.7 g/L) and blood albumin (30.9 ± 5.8 g/L) levels. Gsev showed significant changes in INR (1.74 ± 0.65) and blood sodium (135.3 ± 5.65 mmol/L). If PDI <0.75 was used as the diagnostic criterion for sarcopenia, the incidence was 53.3% in patients weighing >90 kg and 53.6% in those weighing <60 kg. This differed from the PMI, with an incidence of 3.3% in patients weighing >90 kg. Conclusions: The PDI had no significant correlation with body height, body weight or body mass index (BMI) in healthy individuals and patients with ESLD. The PDI was significantly correlated with the severity of cirrhosis and loss of skeletal muscle.

Increased levels of lead in the blood and frequencies of lymphocytic micronucleated binucleated cells among workers from an electronic-waste recycling site.

In recent years, adverse health effects of chemicals from electronic waste (e-waste) have been reported. However, little is known about the genotoxic effects of chemicals in e-waste. In the present study, air concentrations of the toxic metals at e-waste and control sites were analyzed using inductively-coupled plasma mass spectrometry. Levels of toxic metals (lead, copper and cadmium) in blood and urine were detected using atomic absorption spectrophotometry in 48 exposed individuals and 56 age- and sex-matched controls. The frequencies of lymphocytic micronucleated binucleated cells (MNBNCs) were determined using a cytokinesis-block micronucleus assay. Results indicated that blood lead levels were significantly higher in the exposed group (median: 11.449 µg/dL, 1st/3rd quartiles: 9.351-14.410 µg/dL) than in the control group (median: 9.104 µg/dL, 1st/3rd quartiles: 7.275-11.389 µg/dL). The exposed group had higher MNBNCs frequencies (median: 4.0 per thousand, 1st/3rd quartiles: 2.0-7.0 per thousand) compared with the controls (median: 1.0 per thousand, 1st/3rd quartiles: 0.0-2.0 per thousand). Additionally, MNBNCs frequencies and blood lead levels were positively correlated (r = 0.254, p<0.01). Further analysis suggested that a history of working with e-waste was a predictor for increased blood lead levels and MNBNCs frequencies in the subjects. The results suggest that both the living and occupational environments at the e-waste site may be risk factors for increased MNBNCs frequencies among those who are exposed.

Treatment of succinonitrile wastewater by immobilized high efficiency microorganism strains.

Using succinonitrile as a sole source of carbon and nitrogen, two bacterium strains named as J-1-3 and J-13-1 were isolated and screened out from the treatment facilities of Shanghai petrochemical wastewater treatment plant treating acrylic fiber production wastewater. The optimal growth conditions of the two strains in the degradation of succinonitrile with varied initial concentrations were determined through flask tests as follows: temperature 30 degrees C, shaker rotary speed of 250 r/min, inoculum percentage 0.1%, and initial pH 6. Results indicate that the two strains, especially J-13-1, exhibited a high efficiency for succinonitrile degradation. Thereafter, the bacterium strains were immobilized separately by sodium alginate and polyvinyl alcohol 1750 +/- 50, and applied to succinonitrile degradation again. Results show that in a very broad range of the initial succinonitrile concentration, i.e., 30-5,000 mg/L, the sodium alginate immobilized grains could degrade more than 80% of succinonitrile after 24 h under the experimental conditions of 30 degrees C, pH 6.5, and shaker rotary speed of 250 r/min. However, the polyvinyl alcohol immobilized grains tended to inflate and break down due to a weak mechanical strength.

Liver Transplant Recipients With End-Stage Renal Disease Largely Benefit From Kidney Transplantation.

BACKGROUND: The incidence of end-stage renal disease (ESRD) after liver transplant (LT) has increased. The actual benefit of kidney transplantation (KT) is not completely understood in LT recipients with ESRD. METHODS: We analyzed Scientific Registry of Transplant Recipients data for all KT candidates with prior LT from 1998 to 2014; the benefits of KT relative to remaining on dialysis were compared by means of multivariate Cox proportional hazards regression analysis. RESULTS: The number of these KT candidates with prior LT has tripled from 98 in 1998 to 323 in 2015; LT recipients with ESRD remaining on dialysis have a 2.5-times increase in the risk of liver graft failure and a 3.6-times increase in the risk of patient death compared with these patients receiving KT. The adjusted liver graft and patient survival rates after donors from donation after cardiac death or expanded-criteria donor kidney transplantation were significantly higher than in patients remaining on dialysis in LT recipients with ESRD. CONCLUSIONS: The number of referrals to KT with prior LT is increasing at a rapid rate. Remaining on dialysis in LT recipients with ESRD has profound increased risks of liver graft failure and patient death in comparison to receiving a KT. LT recipients with ESRD can benefit from expanded-criteria donor and donation after cardiac death kidney transplantation.

Mass spectrometric screening method for microcystins in cyanobacteria.

A screening method for microcystins in cyanobacteria, which consists of the formation of 3-methoxy-2-methyl-4-phenylbutyric acid as an oxidation product of microcystins by ozonolysis, and detection of 3-methoxy-2-methyl-4-phenylbutyric acid by thermospray-liquid chromatography/mass spectrometry or electron ionization-gas chromatography/mass spectrometry using selected ion monitoring, was developed. The ozonolysis made it possible to significantly reduce the formation times of 3-methoxy-2-methyl-4-phenylbutyric acid because the previously required extraction, clean-up and other procedures could be entirely eliminated. The resulting intact 2-methyl-4-phenylbutyric acid was directly analyzed by thermospray-liquid or electron ionization-gas chromatography/mass spectrometry, and the procedures from ozonolysis to analysis of microcystins at the pmole levels were performed within only 30 min. The calibration curves obtained by thermospray-liquid or electron ionization-gas chromatography/mass spectrometry analysis showed a linear relationship from 14 to 830 pmole and from 2.5 to 100 pmole of microcystin-LR, respectively. The method was applied to the detection and determination of the total amount of microcystins in bloom and cultured samples, showing that it provided a means of not only screening for microcystins but of their accurate quantitation.

[The dissolution rate and physical dispersion state of preparations of biphenyl dimethyl dicarboxylate (DDB)].

DDB is poorly soluble in water. The solid dispersions of DDB with easily soluble carriers such as polyethylene glycol 6000 (PEG 6000), polyvinyl pyrrolidone (PVP) and urea were prepared by melting and solvent methods. The two DDB-PEG 6000 systems are thermodynamically stable interstitial solid solutions. The DDB-PVP system is an amorphous precipitate and the DDB-urea system is a simple eutectic physical mixture judged by X-ray diffraction and thermal analysis methods. The dissolution rate of DDB-PEG 6000 pilule and two kinds of DDB tablets were determined. The dissolution rate of DDB-PEG 6000 pilule was found to be faster. The physical dispersion state is an important factor in relation to the dissolution rate of DDB preparations.

[Bioavailability studies on the preparations of biphenyl dimethyl dicarboxylate(DDB)].

Since the bioavailability of the suspension and the tablet of DDB given orally is only 20-30%, we have prepared four kinds of DDB solid dispersion preparations (DDB pilule I with polyethylene glycol 6000 as the vehicle, DDB pilule II with polyethylene glycol 6000 and absorption accelerator as the vehicle, capsule of DDB-urea fusing mixture and DDB-polyvinyl pyrrolidone coprecipitate), and the bioavailability of these preparations were studied in rabbits, rats and human volunteers by HPLC method. All four preparations showed better absorption than the DDB tablet, and the area under serum DDB concentration-time curve of pilule II was 19 fold that of the tablet in rabbits, meaning that the absorption of pilule II is the best of the four preparations. After administration of the four solid dispersion preparations, the fecal excretion of DDB were all lower than the tablet in both animals and human volunteers. The protective action of pilule II against CCl4 hepatotoxicity was about six times stronger than that of the suspensions. Therefore, there are good reasons to use DDB pilule II instead of the tablets of suspension in the clinic.

Hepatocyte-targeted delivery using ph-sensitive liposomes loaded with lactosylnorcantharidin phospholipid complex: preparation, characterization, and therapeutic evaluation in vivo and in vitro.

Liposomes loaded with lactosyl-norcantharidin phospholipid complex (LPC) were prepared, in which soybean phosphatidylcholine was used to improve the liposolubility of lactosyl-norcantharidin (Lac-NCTD). The pH-sensitive LPC liposomes (pH-LPC-lips) were obtained by electrostatic adsorption of the carboxymethyl chitosan onto the surface of the liposomes. The in vitro drug release of pH-LPC-lips and LPC-lips was investigated in dissolution media with pH ranging from 1.0 to 8.0. The in vitro antitumor activity and cellular uptake of Lac-NCTD and its liposomes to HepG2 cells were studied. The pH-LPC-lips demonstrated strong cytotoxicity against the cells and easily permeated the cell membrane. The in vivo antitumor activities of Lac-NCTD and its liposomes were evaluated in mice bearing H22 liver tumors. The pH-LPC-lips displayed the best tumor inhibitory effect. The optical imaging results indicated that Cy7- labeled pH-LPC-lips showed excellent hepatocyte specificity in H22 tumor-bearing mice. Therefore, pH-LPC-lips can be regarded as liver-targeting agents that combine targeting and active releasing.

Laser photocoagulation treatment of choroidal melanoma.

Ten cases of choroidal melanoma were treated with Argon and/or Krypton laser photocoagulation in this retrospective study. Each patient received an average of 9.6 laser treatment sessions. Three female and seven male patients were treated and followed for an average of 69.9 months. Clinical regression was observed in seven cases (70%). Tumour size in these successfully treated cases ranged from 4.5 x 5.0 x 1.5 mm to 7.0 x 7.0 x 3.5 mm. Continued growth occurred in one case (10%), tumour recurrence which failed to respond to additional laser treatment in another (10%), and although a recurrence appeared to respond to additional laser in a third case (the largest tumour in the series 11.0 x 9.0 x 2.5 mm) extraocular extension was found at enucleation (10%). Complications of laser treatment included cystoid macular oedema (Case 5), branch retinal vein occlusion (Cases 3 and 6), vitreous haemorrhage from neovascularisation at the edge of the scar, optic atrophy and thrombotic glaucoma (Case 8), and macular involvement by laser (Cases 4, 7 and 10). All patients remain alive without any clinical evidence of metastatic disease at the end of the follow-up period. These results suggest that laser photocoagulation treatment is a useful option in the management of small choroidal melanomas. It avoids the psychological trauma of enucleation, and maintains some vision, without compromising life expectancy.

The frequencies and doses of medical exposure in China.

A sampling survey of medical exposure frequency covered 15,000 hospitals in 24 provinces, while a sampling measurement of patient doses covered 2,000 hospitals in 14 provinces. From these surveys and measurements, we compiled survey data of 11 million cases. The data were analyzed by computer, and the frequencies, averages of skin and organic dose per examination, and population doses per capita (He, LSD, GSD, SSD) were found. The possible risks of medical exposure in China are also estimated in this paper.