OBJECTIVE To investigate the effects of celastrol （CSL） on drug resistance of liver cancer cells. METHODS Human liver cancer lenvatinib （Len）-resistant cells Huh7/Len were constructed and divided into control group， CSL low-， medium- and high-concentration groups （1， 2.5， 5 μmol/L）， and CSL high-concentration+Zn27 [focal adhesion kinase （FAK） inhibitor] group （5 μmol/L CSL+2 nmol/L Zn27）， with 6 holes in each group. The proliferation （by absorbance） and cloning ability， apoptotic rate， the number of invasion cells and migration cells， the level of reactive oxygen species（ROS） as well as the protein expressions of phosphorylated FAK （p-FAK）， phosphorylated mitogen-activated protein kinase kinase （p-MEK）， phosphorylated extracellular signal-regulated kinase （p-ERK）， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax） and caspase-3 were detected. RESULTS Compared with control group， cell absorbance， clone count， invasion count and migration count ， and the protein expressions of p-FAK， p-MEK， p-ERK and Bcl-2 were significantly reduced in the CSL low- ， medium- ， high- concentration groups； the apoptosis rate， ROS level， and protein expressions of Bax and caspase-3 were significantly increased， in a concentration-dependent manner （P＜0.05）. Compared with CSL high-concentration group， the changes of above indexes were all reversed significantly in CSL high-concentration+Zn27 group （P＜0.05）. CONCLUSIONS CSL can enhance oxidative stress， promote cell apoptosis， inhibit malignant progression and chemotherapy resistance of liver cancer cells， and its mechanism may be related to the inhibition of the FAK/MEK/ERK signaling pathway.

Objective To explore the value of tuberculosis infected T cells spot test(T-SPOT.TB),heated mycobacterium tuberculosis nucleic acid amplification testing(TB-SAT),and adenosine deaminase(ADA)in diagnosing tuberculous pleural effusion.Methods A total of 135 patients with pleural effusion treated at the First Affiliated Hospital of Xinxiang Medical University from January 2021 to December 2021 were selected as the research subjects,including 83 patients with tuberculous pleural effusion and 52 patients with non-tuberculous pleural effusion.All these patients received peripheral blood T-SPOT.TB,chest water TB-SAT and chest water ADA tests,and the sensitivity and specificity of the above three methods in detecting tuberculous pleural effusion alone and in combination were compared.Results In terms of sensitivity and specificity,there was no statistically significant difference among the T-SPOT.TB,TB-SAT and ADA tests in detecting tuberculous pleural effusion alone(P＞0.05).The sensitivity of the T-SPOT.TB+TB-SAT combined test in detecting tuberculous pleural effusion was significantly higher than that of the T-SPOT.TB,TB-SAT and ADA tests alone(x2=4.990,13.410,14.590;P＜0.05),while the specificity of the T-SPOT.TB+TB-SAT combined test in detecting tuberculous pleural effusion showed no significant difference with that of the T-SPOT.TB,TB-SAT and ADA tests alone(x2=0.000,2.420,0.060;P＞0.05).The sensitivity of the T-SPOT.TB+ADA combined test in detecting tuberculous pleural effusion was significantly higher than that of the ADA test alone(x2=4.069,P＜0.05),but showed no significant difference with that of the T-SPOT.TB and TB-SAT tests alone(x2=0.055,3.384;P＞0.05).The specificity of the T-SPOT.TB+ADA combined test in detecting tuberculous pleural effusion was significantly lower than that of the T-SPOT.TB,TB-SAT and ADA tests alone(x2=4.370,12.511,5.371;P＜0.05).The sensitivity of the TB-SAT+ADA combined test in detecting tuberculous pleural effusion showed no significant difference with that of the T-SPOT.TB,TB-SAT and ADA tests alone(x2=0.000,2.604,3.213;P＞0.05).The specificity of the TB-SAT+ADA combined test in detecting tuberculous pleural effusion was significantly lower than that of the TB-SAT test alone(x2=5.765,P＜0.05),but showed no significant difference with that of the T-SPOT.TB and ADA tests alone(x2=0.782,1.251;P＞0.05).The sensitivity of the T-SPOT.TB+TB-SAT+ADA combined test in detecting tuberculous pleural effusion was significantly higher than that of the T-SPOT.TB,TB-SAT and ADA tests alone(x2=6.760,15.755,16.966;P＜0.05),while the specificity of the T-SPOT.TB+TB-SAT+ADA combined test in detecting tuberculous pleural effusion was significantly lower than that of the T-SPOT.TB,TB-SAT and ADA tests alone(x2=4.370,12.511,5.371;P＜0.05).The sensitivity of the T-SPOT.TB+TB-SAT combined test in detecting tuberculous pleural effusion was significantly higher than that of the T-SPOT.TB+ADA and TB-SAT+ADA combined tests(x2=4.090,4.990;P＜0.05);there was no statistically significant difference in the sensitivity in detecting tuberculous pleural effusion between the T-SPOT.TB+ADA combined test and the TB-SAT+ADA combined test(x2=0.060,P＞0.05).The specificity of the T-SPOT.TB+TB-SAT combined test in detecting tuberculous pleural effusion was significantly higher than that of the T-SPOT.TB+ADA combined test(x2=4.371,P＜0.05);the specificity of the TB-SAT+ADA combined test showed no significant difference with that of the T-SPOT.TB+TB-SAT and T-SPOT.TB+ADA combined tests(x2=0.780,1.490;P＞0.05).There was no statistically significant difference in the sensitivity in detecting tuberculous pleural effusion between the T-SPOT.TB+TB-SAT+ADA combined test and the T-SPOT.TB+TB-SAT combined test(x2=0.210,P＞0.05);the sensitivity of the T-SPOT.TB+TB-SAT+ADA combined test in detecting tuberculous pleural effusion was significantly higher than that of the T-SPOT.TB+ADA and TB-SAT+ADA combined tests(x2=5.750,6.760;P＜0.05).The specificity of the T-SPOT.TB+TB-SAT+ADA combined test in detecting tuberculous pleural effusion was significantly lower than that of the T-SPOT.TB+TB-SAT combined test(x2=4.370,P＜0.05);the specificity of the T-SPOT.TB+TB-SAT+ADA combined test in detecting tuberculous pleural effusion showed no significant difference with that of the T-SPOT.TB+ADA and TB-SAT+ADA combined tests(x2=0.000,1.490;P＞0.05).Conclusion The combined detection performs better than the single detection in diagnosing tuberculous pleural effusion,and the peripheral blood T-SPOT.TB combined with chest water TB-SAT performs the best in detecting tuberculous pleural effusion.The combined detection can effectively reduce the missed diagnosis rate and the misdiagnosis rate,and has high clinical application value for diagnosing tuberculous pleural effusion.

Objective:This paper is to propose a calibration model based on sine function which enables more choices to determine the functional relationship between the absorbance and the concentration of the tested substance.Methods:This paper uses Taylor series expansion and Levenberg-Marquardt to obtain the optimal parameters for the Sine model and then summarizes the characters of the Sine model. On the basis of these characters, this paper compares and evaluates the experimental data processed by the Sine model from four aspects: correctness, precision, linearity and correlation.Results:The generated sine function calibration model achieved deviations within ±3% of the national standard substance, precision ( CV%) less than 2%, and a linear correlation coefficient greater than 0.990 within the measurement range of 32-710 mg/L. The correlation coefficients between the sine model and other well-performing linear calibration models for 104 clinical samples were all greater than 0.990. Conclusions:The performance evaluation of the prealbumin assay kit using the sine function calibration model meets industry standards and shows good correlation with the results of clinical sample measurements. This indicates that the sine function calibration model can serve as a new calibration model for in vitro diagnostic research and clinical applications.

Chronic obstructive pulmonary disease(COPD)is a common respiratory disease with a high disability and mortality rate, and it has become one of the major public health issues of concern worldwide.It includes small airway disease and emphysema caused by abnormal tissue repair, but its pathogenesis is still unclear.COPD is currently viewed as a disease of accelerated lung aging and is closely associated with aging mechanisms such as telomere/telomerase, epigenetic changes and mitochondrial dysfunction.This article reviews research progress on the mechanisms linking COPD and aging-related regulation.

Objective To investigate the relationship between EVI1 gene expression and clinical features and prognosis of children with acute myeloid leukemia (AML). Methods EVI1 gene was detected in AML children, correlation of clinical and lab features, prognosis of AML children with EVI1 gene were analyzed. Results EVI1 expression is positive in 38 of 145 children with AML. There were no significant differences in age, gender, hemoglobin concentration, leukocytes and platelet count, subtype of morphology, ratio of chromosomal anomaly and complex karyotypes between EVI1 positive and EVI1 negative group (P>0.05); coexist genes were detected in 9 cases (23.68%) of EVI1 positive group. Rate of complete remission (CR) was 91.67% in 24 cases of EVI1 positive patients received chemotherapy. Relapse rate was 64.29% and 14.29% in EVI1 positive patients who received chemotherapy and allo-hematopoietic stem cell transplantation (allo-HSCT), retrospectively and significant differences were found (P<0.05). There was no significant difference in CR but significant difference was found in event free survival (P<0.05) for EVI1 positive and EVI1 negative patients who received chemotherapy. EVI1 gene kept negative when bone marrow relapse occurred in two patients with EVI1 positive at diagnosis. Conclusion EVI1 gene may play adverse role in pediatric AML; prognosis of EVI1 positive AML patients can be improved by allo-HSCT; follow-up of EVI1 transcript levels is insufficient to monitoring of minimal residual disease.

Objective Radiological grade of splenic injury was seldomly used in China trauma center now,though it had been established in 1994 by American Association for Surgery of Trauma (AAST) and widely used.The present study is aimed to analyze the imaging grade and clinical characteristics of traumatic splenic rupture in children,discuss the feasibility of conservative treatment,and the role of radiographic grading during clinical treatment.Methods Information (including age,gender,severity based on radiological findings,treatment strategies,and clinical outcome) regarding 59 hospitalized splenic injury patients whose injuries occurred between 2008 and 2014 was retrospectively analyzed.Results Between 2008 and 2014,59 pediatric patients with splenic injury were treated in our institution.Median age was 9.5 years (range,3 months to 16 years).Of all patients,41 (69.5%) were male.The injuries were primarily caused by traffic crash (45.7%),stumbling/falling from a height (38.9%).According to AAST,5 cases were grade Ⅰ,26 patients grade Ⅱ (44.1%),and 21 cases grade Ⅲ (35.6%),6 over grade Ⅳ,and only one was unclear.Of all patients,25 cases were with the other organs complications.All patients underwent fasting,bed rest,and antibiotics.Only 1 case was transferred to operation during the conservative treatment.Forty-nine patients underwent with CT scan over 2 times.Conclusion Imaging classification helps guide clinical treatment.Conservative treatment is feasible for traumatic splenic injury in children.Early imaging classification of splenic injury may be helpful in clinical judgment,and reduce children radiation exposure.

OBJECTIVETo report an acute promyelocytic leukaemia (APL) case with translocation of rob (13;21) t(15;17) (q22;q21) and review its clinical and laboratory characteristics.

METHODSBased on routine karyotype analysis and bone marrow morphology, we further used double color double fluorescent in situ hybridization (DCDF-FISH) and reverse transcriptase PCR (RT-PCR) to examine the patient's abnormities on cytogenetic and molecular biology, and reveal the clinical characteristics of this rare translocation also from the related literatures.

RESULTSThe clinical manifestation and bone marrow morphology examination of this patient were in accordance with pathologic feature of APL. On first visit, immunophenotyping analysis showed positive myeloid markers. Through R-banding, the patient's karyotype was confirmed as 45, XX, rob(13;21) t(15;17) (q22;q21) [6]/45, XX, rob(13;21) [14]. FISH results showed that 68.9% cells were typical t(15;17) pattern. The positive rates of fusion gene of PML-RARα detected by RT-PCR was 25.8%. Patient was treated by induction and consolidation therapy, the karyotype was 45, XX, rob(13;21 )[20] after complete remission. The positive rate of fusion gene of PML-RARα by FISH and its level were 2.5% and 0.003% respectively.

CONCLUSIONAPL with rob (13;21) t(15;17) (q22;q21) was very rare, which was accorded with clinical and laboratory characteristics of APL. The value of chromosome abnormality as a prognostic marker in APL needs to be further observed..

Chromosome Aberrations ; Chromosome Banding ; Chromosomes, Human, Pair 15 ; Chromosomes, Human, Pair 17 ; Chromosomes, Human, X ; Humans ; In Situ Hybridization, Fluorescence ; Karyotype ; Leukemia, Promyelocytic, Acute ; Oncogene Proteins, Fusion ; Remission Induction ; Translocation, Genetic

BACKGROUNDPyruvate phosphate dikinase (PPDK) reversibly catalyzes the interconversion of phosphoenolpyruvate (PEP) and pyruvic acid, leading to catabolism and adenosine triphosphate (ATP) synthesis or gluconeogenesis and ATP consumption. Molecular modeling of PPDKs from divergent organisms demonstrates that the orientation of the phosphorylatable histidine residue within the central domain of PPDK determines whether this enzyme promotes catabolism or gluconeogenesis. The goal of this study was to determine whether PDDK from Giardia underwent adaptive evolution in order to produce more energy under anaerobic conditions.

METHODSA total of 123 PPDK sequences from protozoans, proteobacteria, plants, and algae were selected, based upon sequence similarities to Giardia lamblia PPDK and Zea mays PPDK. Three-dimensional (3-D) models were generated for PPDKs from divergent organisms and were used to compare the orientation of the phosphorylatable histidine residue within the central domain of PPDKs. These PPDKs were compared using a maximum-likelihood tree.

RESULTSFor PPDK from Giardia, as well as from other anaerobic protozoans, the central domain tilted toward the N-terminal nucleotide-binding domain, indicating that this enzyme catalyzed ATP synthesis. Furthermore, the orientation of this central domain was determined by interactions between the N- and C-terminal domains. Phylogenetic analysis of the N- and C-terminal sequences of PPDKs from different species suggested that PPDK has likely undergone adaptive evolution in response to differences in environmental and metabolic conditions.

CONCLUSIONThese results suggested that PPDK in anaerobic organisms is functionally adapted to generate energy more efficiently in an anaerobic environment.

Adenosine Triphosphate ; metabolism ; Evolution, Molecular ; Giardia lamblia ; enzymology ; Protozoan Proteins ; chemistry ; classification ; genetics ; Pyruvate, Orthophosphate Dikinase ; chemistry ; classification ; genetics

Objective To investigate the clinical efficacy of Pingchuan cataplasm combined with hyperthermia therapy for stable chronic obstructive pulmonary disease (COPD) with qi deficiency syndrome of both lung and spleen. Methods Sixty patients of stable COPD with TCM syndrome of qi deficiency of both lung and spleen were selected and randomized into treatment group and control group, 30 cases in each group. Based on conventional treatment, the treatment group was treated by Pingchuan cataplasm combined with hyperthermia, and the control group was given analog stick applicator without additional heat treatment. After 6 weeks of treatment, the TCM symptom score and the time of the first exacerbation after treatment were observed. Results Compared with the control group, Pingchuan cataplasm can reduce the patients’ TCM symptom score (F=9.843, P=0.003), and extend the interval of acute exacerbation after treatment (F=4.451, P=0.040). Conclusion Pingchuan cataplasm combined with hyperthermia applicator can improve symptoms and reduce the frequency of acute exacerbation, thus can treat and prevent the acute exacerbation of COPD.

Objective To summarize the experience of multidisciplinary consultation for prenatal fetal deformity, and to explore the mode suitable for China. Methods The Obstetrics and Gynecology Hospital of Fudan University and Children's Hospital of Fudan University established a joint multidisciplinary consultation center, including obstetrics, pediatrics, pediatric surgery, ultrasound and other departments. A total of 3 378 pregnant women visited the consultation center from July 31, 2003 to August 1, 2013. After consultation, treatment was divided into three classes:pregnancy termination, pregnancy continuation and perinatal treatment. Follow-up was made through correspondence and telephone communication. Retrospective analysis on reasons for consultation, fetal structural abnormalities of the classification system, chronological order of abnormalities, gestational weeks of diagnosis, maternal-related factors, treatment and prognosis was performed. Results (1) Reasons for consultation:Among 3 378 women undertaking prenatal multidisciplinary consultation, 3 243 (96.00%) were due to fetal factors, and 135 (4.00%) were due to maternal factors. (2) Classification of fetal structural abnormalities:Among the 3 243 cases undertaking consultation with fetal factors, fetal abnormality was found in 80.85%(2 622/3 243). The most common were neurological abnormalities(23.19%, 608/2 622), followed by urinary tract malformation (20.25%, 531/2 622) and cardiovascular malformation (15.48%, 406/2 622). These were followed by digestive system malformation, limb deformities and space-occupying lesions. There were 156 cases of multiple malformations. (3) Average gestational weeks for diagnosis of fetal deformity:The 2 622 cases of fetal deformity were diagnosed at a mean (26.7± 2.1) of gestational weeks (21.1–30.4 weeks). Urinary tract malformations were detected at (24.0±0.7) weeks, whereas digestive system malformations were detected at (28.3±2.6) weeks. (4) Induced labor:Induced labor cases accounted for 35.66% (935/2 622), among which, 92 cases were fetal intrauterine death and 843 cases were active choice. The several highest induced labor rates resulted from multiple malformations (75.64%, 118/156), abdominal wall defects (62.22%, 28/45), diaphragmatic hernia (61.54%, 24/39), cleft lip and palate (55.32%, 26/47) and cardiovascular malformations (49.51%, 201/406). For nervous system (27.80%, 169/608), urinary tract (25.80%, 137/531) and digestive system malformations (26.94%, 66/245), induced labor rates were <30%. For abdominal lesions (14.04%, 25/178) and sacrococcygeal teratoma (13.64%, 3/22), induced labor rates were<15%. (5) Continuation of pregnancy in 1 687 cases:Cesarean section was conducted in 1 046(61.94%). Neonatal death occurred in 117(6.94%).(6) Perinatal treatment:Twenty-one cases were treated during pregnancy, including thirteen cases with fetal ascites and hydrothorax treated by drainage, five cases with fetal anemia treated by intrauterine transfusion and three cases with fetal tachycardia treated by digoxin. Ten cases were treated by ex-utero intrapartum treatment. After birth, 297 newborns immediately underwent neonatal surgery. Among these, 259 cases underwent radical surgery, eleven palliative surgery, and sixteen elective surgery after follow-up. Conclusions Prenatal multidisciplinary consultation can make comprehensive multidisciplinary assessment of fetal prognosis and improve the diagnosis and treatment of structural malformations.