[The significance of 8-hydroxy-deoxyguanosine acid in the diagnosis of nonalcoholic steatohepatitis].

Objective: To evaluate the significance of serum 8-hydroxy-deoxyguanosine acid(8-OHdG) in the diagnosis of nonalcoholic steatohepatitis (NASH). Methods: Patients or healthy subjects were enrolled at the Second Hospital of Tianjin Medical University and the Second People's Hospital of Tianjin from May 2013 to December 2015. A total of 41 patients with nonalcoholic fatty liver disease were enrolled in the study, including 20 nonalcoholic simple fatty liver (NAFL) patients and 21 NASH patients whose diagnosis were proven by liver biopsy. The other 32 healthy subjects were studied as controls. Serum 8-OHdG, ALT, AST and GGT were tested. Nonalcoholic fatty liver disease activity score (NAS) and expression of 8-OHdG in liver was investigated between NAFL patients and NASH patients. The correlations between serum 8-OHdG and serum ALT, AST, GGT, and 8-OHdG in liver tissue in NASH group were investigated. In addition, the receiver operating characteristic (ROC) curve analyses for ALT and 8-OHdG levels were performed in NAFL patients and NASH patients, and the cut-off value was determined. Results: Serum 8-OHdG values in healthy controls, NAFL and NASH patients were (0.19±0.16) µg/L, (0.22±0.16) µg/L, (0.42±0.21) µg/L respectively. The serum 8-OHdG and serum ALT, GGT and 8-OHdG in liver tissue were all positively correlated in NASH group with respective correlation coefficient r values as 0.454 7, 0.382 9, and 0.497 6. AUC of 8-OHdG was 0.901 with cut-off value 0.39 µg/L. Its sensitivity was 88.3% and specificity was 81.5%, which were higher than those of ALT. Conclusion: The value of serum 8-OHdG would be used as a marker for the diagnosis of NASH.

Role of IL-10 polymorphisms in susceptibility to hepatitis B virus-related hepatocellular carcinoma.

We conducted a case-control study to investigate the role of three common single nucleotide polymorphisms of IL-10 (-592G/A, -819T/C, and -1082A/C) in the development of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). The study included 173 HBV-related HCC patients and 182 healthy controls. A polymerase chain reaction-restriction fragment length polymorphism assay was applied to assess the sequence variants of interest. Compared with control subjects, HCC patients were more likely to be older (t = 1.94, P = 0.03), have a family history of cancer (chi square = 17.86, P < 0.001), and exhibit higher alanine transaminase (t = 13.32, P < 0.001) and aspartate transaminase (t = 12.63, P < 0.001) levels. Using unconditional logistic regression analyses, we found that the GG genotype of -592G/A was associated with increased risk of HCC [odds ratio (OR) = 2.20, 95% confidence interval (CI) = 1.12-4.38], compared to the AA genotype. Under a dominant model, the AG+GG genotype correlated with HBV-related HCC susceptibility compared to the AA genotype, with an OR (95%CI) of 1.56 (1.02-2.48). Moreover, a recessive model showed the GG genotype to be associated with elevated risk of HCC compared to the AA+AG genotype (OR = 1.85, 95%CI = 1.01-3.47). However, no significant association between the -819T/C and -1082A/C variants and development of HBV-related HCC was observed under codominant, dominant, and recessive models. We conclude that the IL-10 -592G/A polymorphism does play a role in susceptibility to HBV-related HCC under codominant, dominant, and recessive models.

Human chorionic gonadotropin promotes expression of protein absorption factors in the intestine of goldfish (Carassius auratus).

Protein use is crucial for the ovulation and spawning of fish. Currently, limited information is available regarding the expression of protein absorption factors during the breeding seasons of teleosts and thus how various proteins involved in this process is not well-understood. The expression of CDX2, CREB, gluatamate dehydrogenase, LAT2, aminopeptidase N, PepT1, and SP1 were significantly elevated from the non-breeding season to the breeding season in female goldfish, and all proteins except PepT1 and SP1 were elevated in male goldfish. Injection of human chorionic gonadotropin upregulated the expression of all proteins except for aminopeptidase N in female goldfish and SP1 in male goldfish, suggesting a luteinizing hormone-inductive effect on protein absorption factors. Protein use in the intestine is increased during the breeding seasons as a result of increased luteinizing hormone.

Estimate of 2009 H1N1 influenza cases in Shenzhen--the biggest migratory city in China.

The 2009 novel H1N1 influenza pandemic had a significant impact on Shenzhen's population with 2063 laboratory-confirmed human H1N1 cases and five deaths being reported. We used parameters from two population-based surveys and the Shenzhen Influenza Surveillance System to estimate the total number of H1N1 influenza infections in Shenzhen in the 2009 pandemic. The attack rate of influenza-like illness (ILI) in family households was 11·2% (95% CI 9·4-13·0), with 80·2% (95% CI 77·8-82·5) seeking medical care. The ILI attack rate in workers was 38·1% (95% CI 34·3-41·7) with 72·5% (95% CI 66·9-78·0) seeking medical care. The average H1N1 positive rate in individuals reporting ILI and testing by polymerase chain reaction was 22·7%. A total of 611 000-768 000 people, or 4·7-5·9% of the Shenzhen population, are estimated to have experienced H1N1 influenza. The estimated total number of cases of H1N1 is likely to be 330 times greater than the number of laboratory-confirmed cases.

Characterization and ontogenetic expression analysis of the myosin light chains from the fast white muscle of mandarin fish Siniperca chuatsi.

Three full-length complementary DNA (cDNA) clones were isolated encoding the skeletal myosin light chain 1 (MLC1; 1237 bp), myosin light chain 2 (MLC2; 1206 bp) and myosin light chain 3 (MLC3; 1079 bp) from the fast white muscle cDNA library of mandarin fish Siniperca chuatsi. The sequence analysis indicated that MLC1 and MLC3 were not produced from differentially spliced messenger RNAs (mRNA) as reported in birds and rodents but were encoded by different genes. The MLC2 encodes 170 amino acids, which include four EF-hand (helix-loop-helix) structures. The primary structures of the Ca(2+)-binding domain were well conserved among the MLC2s of seven other fish species. The ontogenetic expression analysis by real-time PCR showed that the three light-chain mRNAs were first detected in the gastrula stage, and their expression increased from the tail bud stage to the larval stage. All three MLC mRNAs showed longitudinal expression variation in the fast white muscle of S. chuatsi, especially MLC1 which was highly expressed at the posterior area. Taken together, the study provides a better understanding about the MLC gene structure and their expression pattern in muscle development of S. chuatsi.

[Establishment of a nomogram predicting risk factors of postoperative perineal wound complications after abdominoperineal resection for rectal cancer].

Objective: To investigate the risk factors of perineal incision complications after abdominoperineal resection (APR) for rectal cancer, and to establish a nomogram model to predict the complications of perineal incision. Methods: A case-control study was conducted to retrospectively collect the medical records of 213 patients with colorectal cancer who underwent APR at the First Affiliated Hospital of Nanjing Medical University from January 2010 to December 2016. The complications of perineal incision after APR were classified according to the modified Clavien-Dindo classification of surgical complications (Version 2019), and the complications of grade II and above were defined as "clinically significant complications" .Twenty-two factors related to complication of perineal incision, such as gender, age, surgical procedure, surgical approach, perineal repair, placement of drainage tube, skin position of drainage tube, operation time, intraoperative blood loss, preoperative radiotherapy and chemotherapy, intraoperative local perfusion chemotherapy, tumor classification, pathological grade, tumor T stage, tumor TNM stage and so on, were analyzed by chi-square test for univariate risk factor of complication in all variables, and variables with P<0.2 in univariate analysis were further included in multivariate analysis. Logistic regression analysis was used to screen out independent risk factors. R software (R 3.3.2) was introduced. The rms software package was used to construct a nomogram prediction model. The C-index was calculated (higher meaning better consistency with actual risk) to evaluate the discriminant degree of the model. The Bootstrap method was used to repeat the sampling for internal verification. A total of 42 patients with colorectal cancer who underwent APR from January 2017 to December 2017 at the First Affiliated Hospital of Nanjing Medical University were externally validated, and the corrected C-index was calculated. The model conformity was determined by comparing the C-index calibration difference between the predicted and actual risks. Results: Of the 213 patients with colorectal cancer, 131 were male and 82 were female, with mean age of (59.6±11.6) years. The incidence of postoperative perineal incision complications was 20.2% (43/213), including 27 cases of Clavien-Dindo II and above complications. Univariate analysis showed that the Eastern Cancer Cooperative Group (ECOG) score, preoperative albumin, skin position of drainage tube, intraoperative blood loss, preoperative radiotherapy and chemotherapy were associated with complications of postoperative perineal incision (All P<0.05) . Multivariate analysis showed that preoperative albumin levels ≤38 g/L (OR=105.261, 95% CI: 7.781 to 1423.998, P<0.001), perinead drainage (OR=11.493, 95% CI: 1.379 to 95.767, P=0.024), intraoperative blood loss >110 ml (OR=6.476, 95% CI: 1.505 to 27.863, P=0.012) and preoperative radiotherapy and chemotherapy (OR=7.479, 95% CI: 1.887 to 29.640, P=0.004) were postoperative clinically significant independent risk factors for perineal incision complications. The nomogram model was established. Preoperative albumin level <38 g/L was for 100 points, the preoperative chemoradiotherapy was for 52.5 points, the intraoperative blood loss >110 ml was for 28.5 points, and the perineal drainage was for 17.5 points. Adding all the points was the total score, and the complication rate corresponding to the total score was the predicted rate of the model. The model had a C-index of 0.863. After internal verification, the C-index dropped by 0.005. External verification showed a C-index of 0.841. Conclusions: Preoperative nutritional status, skin position of drainage tube, intraoperative blood loss and preoperative radiotherapy and chemotherapy may affect the occurrence of perineal wound complications after APR for rectal cancer. The nomogram model constructed in this study is helpful for predicting the probability of clinically significant complications after APR.

Autophagic and apoptotic cell death in amniotic epithelial cells.

The aim of this paper is to determine if autophagic cell death is associated with apoptosis and whether it participates in the process of term amniotic rupture. Forty pieces of fresh term amnions, including twenty from a position near the margin of the placentas and twenty from the margin of the naturally ruptured part of the placentas in term gestation were collected, respectively. The amnions were examined by scanning electron microscopy (SEM) and amniotic epithelial (AE) cells were examined by transmission electron microscopy (TEM). Autophagic and apoptotic cell death (PCD) were assayed by laser scanning confocal microscopy (LSCM) or flow cytometry using monodansylcadaverin (MDC) and propidium iodide (PI) stain. BCL(2) and BAX were examined by immunoblotting. Under SEM the amniotic epithelia appeared normal in the position near the placenta. They had an atrophied appearance in the margin of their natural broken parts. In the AE cells PCD was divided into three subtypes by TEM: autophagic cell death with positive stains of MDC and PI; apoptotic cell death; and the mixed type. Quantitative detection showed that there were more death cells, including autophagic and apoptotic, in the AE cells near the ruptured parts than near the placentas. An increased expression of BAX and a decreased expression of BCL(2) protein in the AE cells near the broken margin were observed. Apoptotic and autophagic cell death by the intrinsic pathway are the basic event in the AE cell and they are involved in the cause of membrane rupture of the human amnion in term gestation.

2-(3',5'-Dimethoxybenzylidene) cyclopentanone, a novel synthetic small-molecule compound, provides neuroprotective effects against ischemic stroke.

2-(3',5'-Dimethoxybenzylidene) cyclopentanone (DMBC) is a novel small-molecule compound synthesized by our group. Here, we found that in rat models of permanent middle cerebral artery occlusion (pMCAO), intraperitoneal injection (ip) of DMBC at 1h after ischemia reduced infarct volume, improved neurological deficits and increased the protein levels of microtubule-associated protein 2 (MAP 2) and glial fibrillary acid protein (GFAP) in the ischemic cortex. Post-treatment of DMBC still produced neuroprotective effects even when administered at 6h after ischemia. In the oxygen-glucose deprivation (OGD)-induced astrocytes or HT22 cell injury, DMBC treatment decreased the OGD-induced lactate dehydrogenase (LDH) leakage and increased the GFAP levels in astrocytes. In addition, Annexin-V-Fluos staining analysis revealed that DMBC treatment attenuated both OGD-induced apoptosis and necrosis in astrocytes. Western blotting analysis showed DMBC treatment inhibited the ischemia or OGD-induced increases in active cathepsin B in the ischemic cortex or in astrocytes or HT22 cells. Immunofluorescence analysis demonstrated that DMBC treatment blocked the ischemia or OGD-induced release of cathepsin B from the lysosomes into the cytoplasm in the ischemic cortex or in astrocytes or HT22 cells. Taken together, our results indicate that DMBC can offer neuroprotective effects against cerebral ischemia with an extended therapeutic window and its mechanism might be associated with inhibition of the cathepsin B activation.

Cytokine expression during acute graft-versus-host disease after allogeneic peripheral stem cell transplantation.

Acute graft-versus-host disease (aGVHD) is one of the major problems following allogeneic peripheral blood stem cell transplantation (allo-PBSCT). In order to investigate the pathogenesis of human aGVHD, we analyzed cytokine gene expression and cytokine secretion by peripheral blood mononuclear cells (PBMC) in 30 patients who underwent allo-PBSCT. In this study, reverse transcription-polymerase chain reaction (RT-PCR) was used to explore the mRNA expression of interleukin (IL)-2, interferon (IFN)-gamma, IL-4, IL-10 and IL-12 in the PBMC of allo-PBSCT patients with aGVHD and in controls. The concentrations of these cytokines and of IL-18 were also measured by means of ELISA in medium obtained from cultured leukocytes after stimulation with PHA or LPS. Compared with the normal allo-PBSCT group (n=14), IL-2 and IFN-gamma were detected more frequently in aGVHD group (n=16). IL-12 and IL-18 were elevated, while IL-10 level decreased in the same group. There is no difference in IL-4 gene expression between patients with or without aGVHD, but the concentration of IL-4 for patients with grade II-IV aGVHD decreased. These data suggest that IL-2, IFN-gamma, IL-12 and IL-18 play important roles in the development of aGVHD in humans.

Molecular comparison of Giardia lamblia isolates.

Giardia lamblia (also Giardia duodenalis, Giardia intestinalis) isolates have been variably divided into two or three genotypes by different investigators. We have compared the triose phosphate isomerase sequences of the three genotypes (Groups 1, 2, and 3) described by Nash and shown that Groups 1 and 2 are similar, while Group 3 is markedly different from Groups 1 and 2, indicating that Group 1/2 and Group 3 correspond to the two major genotypes identified by other investigators. We have also analysed three Chinese isolates and showed that two fit into Group 3, while the third contained a mixture of Groups 1 and 3 isolates. These results confirm the relatedness of G. lamblia isolates from throughout the world, and established the feasibility of using DNA amplification and sequence analysis for detecting mixed isolates.

Four isolates of Giardia lamblia cultivated axenically in China and the restriction endonuclease analysis of their DNA.

Axenic culture of 4 Giardia lamblia isolates, 3 from humans and 1 from a rabbit, have been established in China for the first time. The trophozoites of Giardia used for the initial cultivation were obtained from the purified cysts excysted in suckling gerbil (Meriones unguiculatus) and BALB/c mice. Modified TYI-S-33 medium was used in establishment of the culture. DNA banding patterns were compared using restriction-endonuclease analysis and then followed by Southern blot analysis with genomic DNA as a probe. The 4 isolates showed identical banding patterns after being digested with Hind III, Bgl II, Pts I, Hae III, and Hinf. Two different banding patterns were observed after further digestion with Alu I and southern blot analysis. This indicated that isolates of human and rabbit origins do not have strict host specificity and that cross transmission may occur between humans and rabbits.

Establishment of an axenic culture of Giardia lamblia through preliminary passage in suckling gerbil.

Cysts of Giardia lamblia from fresh stool specimens of a 13-year-old boy, who lived in suburban Beijing and had suffered from recurrent diarrheas for years, were concentrated, purified and suspended in physiological saline. Six suckling gerbils (Meriones unguiculatus) were each inoculated with 0.2 ml of this suspension which contained 20,000 cysts/ml. On day 8 after inoculation, the infected animals were sacrificed and trophozoites of G. lamblia which appeared numerous in the upper small intestines were isolated aseptically. The trophozoites were inoculated into modified TYI-S-33 medium and cultivated at 37 C. On day 14 of cultivation, the organism grew luxuriantly and formed a cellular monolayer on the inner wall of the culture tube. The organism replicated every 15 +/- 2 hours and the peak growth occurred 120 hours after seeding. No bacterial growth was detected by repeatedly transferring the culture onto blood agar plates and into beef broths. Several lots of the culture have been subjected to cryopreservation in liquid nitrogen and the viability of the thawed organism ranged from 50% to 80% 1-10 weeks after being cryopreserved. The thawed parasites grew well in subcultures. This axenic culture has been maintained for one year and 4 months, and more than 150 subcultures have been made.

Detecting giardia lamblia antigen in fecal matter with counterimmuno-electrophoresis in diagnosis of giardiasis.

Feces specimens from 35 patients with parasitologically confirmed giardiasis, 41 with acute gastroenteritis, 23 with acute bacillary dysentery, 40 normal persons as well as 15 jirds experimentally infected with Giardia lamblia were used for detecting parasite antigen by counterimmunoelectrophoresis (CIE). It revealed that 33 (94%) of the 35 patients with giardiasis and 14 (93%) of 15 infected jirds were positive, while the other cases, either normal person or normal jird were negative. CIE was also performed in 4 with giardiasis before and after metronidazole treatment. Prior to metronidazole administration, they were all CIE positive, but from the second day on, all became CIE negative. Apparently, detecting giardia antigen in feces specimens by CIE is not only sensitive in detecting current infection but also helpful in evaluating the effects of antigiardia agents on giardiasis.

Colon replacement from esophagus. Clinical experience from 240 cases.

The clinical experience in the colon replacement of the esophagus in 240 cases is reported. The overall operative morbidity was 17.5%; the incidence of the anastomotic leaks was 10.4%; and the mortality was 2.80%. The points in surgical technique were: 1) utilizing the left colic artery as the supporting vessel if possible, based on the anatomy of the colon vessel; 2) using the colon segment in an isoperistaltic position, which has been proved much physiological; 3) single-layer anastomosis which is simple and reliable, with minimal inflammation and quick healing; and 4) choosing the channel of the colon transplant according to the disease condition, the age, and the function of the heart and the lung of the patients.

[The characteristics of Bacillus thuringiensis strain YBT833 and its transformants that containing different ICP genes].

Four different transformants were selected by transferring cry1C into Bacillus thuringiensis strain YBT833. Southern blot and Plasmid profiles results all proved that cry1C was transferred into strain YBT833. However, it was found by PCR analysis that transformant YBT833-1 kept all indigenous ICP(insecticidal crystal protein) genes of strain YBT833 while transformant YBT833-2 lost cry1Ab, and transformant YBT833-3 lost all ICP genes. SDS-PAGE showed that transformants of YBT833-1, YBT833-2 and YBT833 all had 130 kDa and 65 kDa peptide bands, but transformant YBT833-3 did not have 65 kDa peptide band. Bioassay results showed that the toxicities of all transformants to Spodoptera exigua, Heliothis armigera and Plutella xylostella were lower than that of strain YBT833, except the toxicity of transformant YBT833-2 to P. xylostella which was obviously higher than that of YBT833.

[Phylogenetic relationships among Viperidae, Crotalinae based on mitochondrial 12S rRNA sequence variations].

This paper analyzed the phylogenetic relationships and classification of pit vipers (Viperidae, Crotalinae) based on mitochondrial 12S rRNA gene sequence variations. We have sequenced mtDNA 12S rRNA gene about 370 bp fragment from Gloydius saxatilis Emelianov, Gloydius shedaoensis Zhao, Gloydius ussurriensis (Emelianov), Trimeresurus stejnegeri Schmidt and Deinagkistrodon acutus, Gloydius brevicaudus (Stejneger) from two different localities, respectively. Combined with the sequence of Dinodon semicarinatus from GenBank, we have constructed two molecular phylogenetic trees using both Maximum-parsimony analysis and Neighbor-joining. Our results support the following conclusions: (1) Deinagkistrodon is a valid genus; (2) we also discussed the origin of Gloydius shedaoensis and showed it is a subspecies of Gloydius saxatilis; (3) Gloydius brevicaudsu and Gloydius ussurriensis are classified two species.

[Excretion of trichloroacetic acid (TCA) in urine of workers exposed to trichloroethylene].

The excretion pattern of TCA in urine of workers exposed to trichloroethylene and the correlation between concentration of TCA in urine and uptake amount of trichloroethylene were investigated by applying Head Space Gas Chromatography. We examined 33 unexposed subjects (control group) and 48 workers (exposure group). The results showed that the concentration of TCA in urine rose obviously after exposure to trichloroethylene. The concentration of TCA in urine and trichloroethylene exposure was positively correlated (X = 2.916Y + 4.619, r = 0.956 8). Based on the observation of 6 volunteers, we found that the concentration of TCA in urine reached its peak about 48 hours after exposure has ceased. It then slowly declined and did not recover completely up to 96 hours.

[Comparative effects of carvedilol and metoprolol in preventing from left ventricular remodeling after acute myocardial infarction in rats].

OBJECTIVE: To compare the effects of carvedilol and metoprolol in preventing from left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in rats. METHODS: Twenty-four hours after ligating left coronary artery, 105 surviving female SD rats were randomly assigned to AMI control, carvedilol 1 mg/(kg.d) and metoprolol 2 mg/(kg.d) groups. Sham-operated rats (n = 16) were selected randomly as non-infarction control. After four weeks of drugs therapy, hemodynamic studies and pathologic analysis were performed. Exclusive of MI size < 35% or > 55%, complete experimental variables were obtained in 46 rats, which were comprised of AMI (n = 11), carvedilol (n = 12), metoprolol (n = 11), and Sham-operated (n = 12) groups. RESULTS: Compared with sham-operated group, left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV) and weight (LVW), were all significantly increased (P < 0.05-0.001), while maximal rate of rise and fall (+/- dp/dt) of LV pressure as well as their corrected values (+/- dp/dt/LVSP) were all significantly decreased (P < 0.01-0.001) in AMI group. In comparison with AMI group, the LVEDP and LVV were all significantly decreased (all P < 0.001), while +/- dp/dt and +/- dp/dt/LVSP were significantly increased (P < 0.05-0.001) in both carvedilol and metoprolol group, with LVW and RVW only decreased in carvedilol group (P < 0.05-0.01). CONCLUSIONS: 1. Carvedilol can effectively attenuate LVRM, and improve hemodynamics and LV function after AMI in rats, 2. Metoprolol has equivalent beneficial effects as carvedilol on hemodynamics, LV dilatation and function, but not LV hypertrophy.

[Children's serum hepatitis after open heart surgery: follow-up of 230 patients].

From 1984.4. to 1989.5, 230 children with congenital heart diseases underwent open heart operation. (mean age: 6.51 +/- 2.94 years, mean body weight: 18.81 +/- 6.71 kg, mean transfused blood volume: 1353 +/- 277 ml.) Follow-up study revealed that 11 patients developed serum hepatitis after surgery. The difference of incidence between our group (4.78%) and natural group (0.039%) is significant (P less than 0.001). Statistical analysis showed that the incidence of serum hepatitis after operation does not depend on the volume of blood transfused (P greater than 0.50). The authors believe that certain preventive measures should be taken.