Objective To explore the role of granzyme B( GzmB) in infiltrated myocardial cells of mice with viral myocarditis( VMC) induced by coxsackievirus B3(CVB3) during the injury of myocardium and further to elucidate the pathogenesis of VMC. Methods Thirty-six BALB/c male mice, 5 to 6 weeks old, were randomly divided into two groups: the normal group, 10, and the model group, 26. On the eleventh day all mice were killed and the hearts were taken out for inspection whether there were pathological changes in the myocardium by macroscopy and microscopy. The expression of GzmB was detected in myocardium with immunohistochemistry and hemi-quantitive reverse transcript-polymerase chain reaction ( RT-PCR). Results GzmB positive cells in the mice myocardial cells infiltrated were detected by immunohistochemistry in the model group,and the number of GzmB positive cells is remarkable positive correlation to myocardial pathological score(r =0. 859, P =0.000) , however, no such cells were discovered in the normal murine hearts. Test of RT-PCR showed markedly high expression of GzmB mRNA in mice myocardium of the model group, however, no expression of GzmB mRNA in the normal group. Conclusion The expression of GzmB in the infiltrating myocardiocytes probably plays some role during the injury of myocardium, which mediated cytotoxicity effect is one of important mechanisms of myocardial injury in viral myocarditis. The expression quantity of GzmB in the infiltrating myocardiocytes is significant positive correlation to the severity of myocardial injury.

Humans ; Phosphorus Compounds ; toxicity ; Poisoning ; therapy

Objective To construct a subtractive cDNA library of genes transactivated by XTP3 protein with suppression subtractive hybridization technique in order to clone genes associated with transactivation. Methods mRNA was isolated from HepG2 cells transfected by pcDNA3.1(-)-XTP3 and pcDNA3.1(-) empty vector respectively, and then cDNA was synthesized. After restriction enzyme Rsa Ⅰ digestion, small sizes cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2 respectively. After being hybridized with driver cDNA twice and underwent two times of nested PCR, tester cDNA was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.Coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. Results The amplified library contained 30 positive clones. Colony PCR showed that 23 clones contained 200-1 000bp inserts. Sequence analysis was performed also. It was found that 20 kinds of known and 2 kinds of novel cDNA sequences may be target genes transactivated by XTP3 protein. Conclusion The subtractive library of genes transactivated by XTP3 protein was constructed successfully. It provides a foundation for the further research on pathogenesis of the viral proteins.

Objective To investigate the correlation between serum soluble epithelial cadherin (sE-cad) and postoperative recurrence and prognosis in patients with advanced gastric cancer.Methods The level of serum sE-cad in 85 patients with advanced gastric cancer (advanced gastric cancer group) was detected by ELISA technique preoperative and postoperative 1 month,and compared with 30 healthy controls(control group).The patients in advanced gastric cancer group were followed up for 3 years,the level of serum sE-cad in recurrent patients and non-recurrent patients was compared.Results The level of serum sE-cad in advanced gastric cancer group preoperative was significantly higher than that in control group [(24.3 ± 14.8) μ g/L vs.(9.4 ± 3.8) μ g/L,P ＜ 0.01].The level of serum sE-cad in advanced gastric cancer group was significantly decreased postoperative 1 month [(12.5 ± 6.4) μ g/L vs.(24.3 ± 14.8) μ g/L,P ＜0.01].The level of serum sE-cad in recurrent patients was significantly higher than that in non-recurrent patients and postoperative 1 month [(20.7 ±9.8)μg/L vs.(12.5 ±6.4),(14.8 ±6.2) μg/L,P＜0.01].Univariate analysis revealed that preoperative high serum sE-cad level was related with tumor size,differentiated degree,lymph node metastasis ratio,depth of tumor invasion (P ＜0.05),but had no relationship with histological type(P＞ 0.05).Elevated preoperative serum sE-cad level negatively affected the postoperative survival rate and recurrence rate.Multivariate Logistic regression anaiysis revealed that preoperative serum sE-cad level was an independent risk factor for postoperative 3 years survival rate in advanced gastric cancer (HR =2.068,P =0.013).Conclusions Preoperative elevated serum sE-cad level is related with pathologic features in patients with advanced gastric cancer,and may be an important prognostic factor.Postoperative monitoring the level of serum sE-cad is useful for evaluating the prognosis and recurrence.

Abstract: Objective　To understand the characteristics of mutations associated with resistance among 72 multidrug-resistant tuberculosis (MDR-TB) strains using whole genome sequencing (WGS) and to evaluate the performance of WGS for predicting MDR-TB drug resistance. Methods　The clinical strains isolated from patients who visited the outpatient department of Tianjin Center for Tuberculosis Control from January to September in 2020 were collected. Identification tests using p-nitrobenzoic acid (PNB) medium were performed. Drug susceptibility tests (proportion method) on L-J medium were performed. After excluding duplicate strains, 72 MDR-TB strains were selected for WGS. Data were analyzed by using online databases and the phenotypic drug susceptibility test results were compared with resistance profiles predicted by WGS. Results　All of 72 MDR-TB strains belonged to linage 2, and there was no significant difference in rate of pre-extensive drug-resistant tuberculosis (pre-XDR-TB) between modern type and ancestral type (χ2=0.287, P=0.592). A total of 81 mutation types were found from resistance-related genes for 12 anti-tuberculosis drugs, and the common mutation types in different drug-resistant strains were: streptomycin (SM): rpsL Lys43Arg; isoniazid (INH): katG Ser315Thr; rifampicin (RIF): rpoB Ser450Leu; ethambutol (EMB): embB Met306Val; ofloxacin (OFX), levofloxacin (LFX), moxifloxacin (MFX): gyrA Asp94Gly; kanamycin (KAM), capreomycin (CAP), amikacin (AMK): rrs 1401a>g; para-aminosalicylic acid (PAS): folC Ile43Thr. Nine mutation types were found in 9 prothionamide (PTO)-resistant strains, one type for each strain. The sensitivity and specificity of WGS for predicting resistance to different drugs were SM: 98.15% and 88.89%, INH: 90.28% and -, RIF: 98.62% and -, EMB: 79.49% and75.76%, OFX: 97.30% and 85.71%, KAM: 85.71% and 98.46%, PAS: 27.27% and 95.08%, PTO: 81.82% and 60.66%, CAP: 60.00% and 98.51%, LFX: 97.22% and 83.33%, MFX: 97.30% and 85.71%, AMK:100.00% and 100.00%, respectively. Conclusion　WGS is a rapid and promising method which has high consistency with the phenotypic drug sensitivity test. Therefore, it has good application prospects in predicting drug resistance in MDR-TB.

Abstract： Objective　To analyze the species distribution of microorganisms in culture tubes reported positive byBACTEC™ MGIT960 (hereafter referred to as "MGIT960") after species identification using matrix-assisted laser desorption/ionization time of flight massspectrometry (MALDI-TOF MS) technique. Methods　From 2021 to 2022, a total of 2 662 positive tubes reported by the MGIT 960 instrument at Tuberculosis Reference Laboratory of Tianjin Center for Tuberculosis Control were collected. Liquid cultures were independently inoculated to blood plate and neutral L-J medium, and the resulting isolate strains were identified using the MALDI-TOF MS method. According to the MALDI-TOF MS results, the non-repetitive results of the same patient on the same culture medium were analyzed for the composition ratio of strain distribution. For the strains not identified by MALDI-TOF MS, 38 strains were selected for 16S rRNA gene sequencing. Results　A total of 605 isolates were obtained from blood plates, and 501 of those were analyzed. Among them, Mycobacterium accounted for 17.76% (89/501), predominant by Mycobacterium abscess 10.18% (51/501) and Mycobacterium fortuitum 3.19% (16/501). Bacteria other than Mycobacterium accounted for 68.06% (341/501), with the main ones being Nocardia farcinica 15.57% (78/501), Gordonia sputa 9.38% (47/501) and Gordonia bronchialis 7.58% (38/501). There were 71 unidentifiable strains, making up 14.17% (71/501). A total of 2 378 strains were isolated from neutral L-J mediums, 1 748 of which were used in the incoming analysis. Among these, 78.72% (1 376/1 748) were Mycobacterium, 60.53% (1 058/1 748) were Mycobacterium tuberculosis (MTB), 4.69% (82/1 748) were Mycobacterium chimaer intracellulare group and 3.55% (62/1 748) were Mycobacterium Lentiflavum. Bacteria other than Mycobacterium accounted for 17.11% (299/1 748) in neutral L-J medium isolates, with Nocardia farcinica 4.35% (76/1 748), Gordonia sputa 2.69% (47/1 748) and Gordonia bronchialis 2.12% (37/1 748) as the main species. There were 73 strains that couldn't be identified, comprising 4.18% (73/1 748). The 38 strains that not identified by MALDI-TOF MS were all found to be Actinobacteria and Firmicutes by sequencing. Conclusions　A variety of nontuberculous Mycobacterium and bacteria other than Mycobacterium were found in the positive culture tubes reported by the MGIT960 instrument, most of which could be quickly identified by mass spectrometry. Bacteria other than Mycobacterium are mainly Nocardia and Gordonia, which should be paid attention to in differential diagnosis.

Fifteen patients with chronic renal failure(CRF)underwent laparoscopic cholecystectomy(LC)at carbon dioxide(CO2)pneumoperitoneum pressure of 10-12 mm Hg(Group A,n=9)or 13-15 mm Hg(Group B,n=6).Renal function and urinary volume(UV)of Group A showed no remarkable change following the operation.But in Group B,the levels of blood urine nitrogen(BUN)and serum creatine(Scr)were increased significantly,and creatinine clearance rate(Ccr)and UV were remarkably decreased(P＜0.05).These variants gradually retumed tO the preoperative levels after 1 week.The analysis showed that laparoscopic choleeystectomy at CO2 pneumoperitoneum pressure of 10-12 mm Hg in CRF patients might be safe.Higher CO2 pneumoperitoneum pressure could result in reversible renal Email:yuanhuizhong2000@yahoo.com.cnfunction change.

Objective To explore the impairment of time perception in patients with Parkinson's disease (PD).Method Thirty-two individuals with idiopathic PD were compared with 32 matched healthy controls using a time reproduction task.Result Compared with healthy controls for the 3-second duration reproduction task, PD patients in the setting of a delay by 1 s ( (2.78 ±0.31 ) s and (2.15 ±0.34) s,respectively) and a delay by 5 s ( (2.74 ±0.32) s and (2.08 ±0.37) s, respectively) showed impairment in time perception with a significant under-estimation of the duration ( t = - 7.459, - 7.533, both P ＜0.01 ).When compared with healthy controls for the 5-second duration reproduction task, PD patients in the setting of a delay by 1 s ( (3.99 ±0.39) s and (3.11 ±0.52) s) and 5 s ( (4.05 ±0.40) s and (2.96 ±0.54) s) also showed impairment in time perception with a significant under-estimation of the duration (t =-7.609 and - 9.120, both P ＜ 0.01 ).When compared with healthy controls for the 600-millisecond duration reproduction task, PD patients in the setting of delay by 1 s ((0.91 ±0.18) s and (0.90 ±0.18) s, respectively) and by 5 s ( (0.89 ± 0.16) s and (0.91 ± 0.17 ) s, respectively) did not have impaired time perception, and the difference was not significant ( t = 0.347, P = 0.730; t = - 0.519, P =0.606, respectively).Conclusion Our data indicates that time perception is impaired in PD patients.Impaired time perception is mainly an under-estimation of the time interval in seconds rather than milliseconds.

Objective To discuss the surgical treatment of spinal injury, and provide insights on key points and related issues for operations. Methods Two hundred and five cases of spinal fracture were treated through posterior surgical treatment. Under C-arm X-ray monitoring, surgeries had been operated on pedicle screws insertion, vertebral canal decompression, over-extending reduction position, and placed the connecting rod, knocked in the bone graft and finally transplanted the paraspinal bone. Results After operations , the height and morphology of vertebral bodies and spinal physiological curvature were basically recovered analyzed by X ray examination. The follow-up results (in the average of 14 to 36 months) indicated that there were 4 cases of delayed infection, 7 cases of loosen screw, 6 cases of broken screw (14 screws)and 1 case of broken stick, with no secondary nerve injury or other syndromes. Conclusion The vertebral pedicle screw internal fixation manipulated easily, which could sufficiently enlarge vertebral canal in order to decompression. In addition, during the operation, together with over-extending reduction position is beneficial to regain the height of fractured vertebral bodies.

Objective:To explore the influence of selective fetoscopic laser occlusion of chorioangiopagous vessels (FLOC) versus Solomon surgery in the management of twin-to-twin transfusion syndrome (TTTS) on procedure-related complications, fetal survival rate, and residual anastomoses.Methods:A total of 59 pregnant women with TTTS who underwent FLOC in the Third Affiliated Hospital of Zhengzhou University from May 2018 to March 2020 were retrospectively enrolled and divided into Solomon ( n=33) and selective FLOC groups ( n=26) based on the FLOC operation method. Placentae of 25 pregnant women (15 in the Solomon group, 10 in the selective FLOC group) with both survival twins were perfused to observe the type and diameter of the residual anastomoses. Fetal survival rate, procedure-related complications, and the type and diameter of residual anastomoses were analyzed and compared between the two groups using two independent samples t-test, Mann-Whitney U test, χ2 test and Fisher's exact test. Results:(1) The operating time of Solomon was shorter than that of selective FLOC [74 min (60-90 min) vs 95 min (81-123 min), Z=2.906, P=0.004]. But no statistically significant differences in the gestational week at operation, time of pregnancy end, and the interval between operation and pregnancy end was observed between the two groups (all P>0.05). (2) There was no statistically significant difference in the survival rate of both twins, one fetus, at least one fetus, and the incidence of postoperative twin anemia-polycythemia sequence (TAPS) and recurrent TTTS in the Solomon group and selective FLOC group [64%(21/33) vs 50%(13/26), χ2=1.107; 15%(5/33) vs 35%(9/26), χ2=3.044; 79%(26/33) vs 85%(22/26), χ2=0.326; 3%(1/33) vs 12%(3/26), χ2=1.368; 0% (0/33) vs 4%(1/26), χ2=1.118; all P>0.05]. (3) There was no statistically significant difference in the number of placentae with residual anastomoses or the number of artery-to-vein, vein-to-artery, artery-to-artery, and vein-to-vein anastomoses between the two groups (7/15 vs 6/10, 2/8 vs 4/15, 3/8 vs 4/15, 2/8 vs 5/15, 1/8 vs 2/15; Fisher's exact test, all P>0.05), but the diameter of the residual anastomoses in the Solomon group was smaller than that in the selective FLOC group [(0.8±0.3) and (2.2±0.7) mm, t=0.764, P=0.034]. (4) Among the four pregnant women developed TAPS after operation, one patient had two residual artery-to-vein anastomoses in the placenta with diameter of 0.54 mm and 0.43 mm, respectively; one patient had one artery-to-vein anastomosis with a diameter of 0.64 mm; one had one artery-to-artery and one vein-to-vein anastomosis with diameter of 1.56 mm and 1.89 mm, respectively. Conclusions:Compared with selective FLOC, Solomon surgery can reduce the vessel diameter of residual anastomoses in women with TTTS, but does not reduce postoperative complications, nor improve the pregnancy outcomes.