Objective The studies on tissue culture and cytological observations of leaf explants of Curculigo orchioides were conducted in order to provide the basis for the rapid propagation of C. orchioides. Methods Young leaf explants of C. orchioides were cultured on MS basal media. Differences in the callus induction and plantlet regeneration rate were observed by different light treatment as well as chemical factors like different phytohormones, casein hydrolysate (CH), and activated charcoal (AC) concentrations. Paraffin method was used to cytological observation. Results For callus induction of leaf explants of C. orchioides, dark treatment gave better results compared to light treatment; among the media tested, the suitable phytohormone combinations were 2.0 mg/L 2, 4-D or 6-BA 1.5 mg/L+2, 4-D 2.5 mg/L, and 300 mg/L CH+0.2% AC was good for plantlet regeneration from leaf explants. The callus from leaf explants mainly originated from midrib. The parenchyma cells near epicuticle of midrib firstly were initiated to division. Then the parenchyma cells of vascular bundle sheath and mesophyll cells on each side of vascular bundle were also divided to form callus. The buds developed on the peripheral parts of the calli, but the roots developed in the regions deep within the calli. Conclusion Tissue culture of young leaf explants of C. orchioides can make the propagation of C. orchioides rapid.

Objective: To observe the changes of the cognitive function of patients with polycystic ovary syndrome(PCOS) and explore their relationship with serum testosterone.Methods: The levels of serum testosterone(T),dehydroepiandrosterone sulfate(DHEAS),sex hormone binding globulin(SHBG),Estradiol(E2),follicle-stimulating hormone(FSH) and luteotrophic hormone(LH) were measured by radioimmunity assay in 25 patients with PCOS and 25 normal women.Their cognitive functions were assessed by delayed word recall test,word learning test,symbol digit substitution test,animals category fluency test,block design,trail making test(Part A) and digit span test.And the results of the tests were compared between the PCOS and the normal group.Results: The levels of serum T,LH and SHBG in the PCOS patients differed significantly from the normal individuals(P

Objective To detect the isoniazid (INH) and rifampin (RIF) resistance of Mycohaeterium tuberculosis isolates in the single tube with multiplex-polymerase chain reaction-single strand conformation polymorphism(muhi-PCR-SSCP) system. Methods According to the sequences of inhA, katG and rpoB genes of the Mycohacterium tuberculosis, three pairs of oligonucleotide primes were designed to examine the INH and RIF resistance with the multi-PCR-SSCP. The validity of the newly developed method was evaluated with 116 clinical isolates of Mycohacterium tuberculosis( 70 isolates that were INH-resistant and 66 isolates that were RIF-resistant). Results The validity of the method was assessed with multiplex PCR-SSCP with the bacteria culture with susceptibility test as golden standard. The three genes, katG, inhA and rpoB, in the 116 clinical isolates and H37Rv strain were amplified successfully in single PCR reactions,except 4 isolates with katG deletion mutants. Compared with strain H37Rv, forty-six isolates had katG gene mutations, thirteen had inhA mutations and fifty-eight had rpoB mutations. Thirty-eight isolates had simultaneous katG and rpoB mutations and 4 isolates had both inhA and rpoB mutations. Four isolates had inhA and katG mutations and 2 isolates had mutations in all three genes simultaneously. The sensitivity of the newly developed multiplex-PCR-SSCP assay was 80% and 82% for INH and RIF, respectively. The specificity of the assay was 100% and 92% for INH and RIF, respectively. Conclusion Muhiplex-PCRSSCP provides a rapid, specific and cost-effective method of detecting multidrug-resistant TB. It laid a solid foundation for the further study of drug resistant gene.

Objective To investigate the feasibility of the simultaneous measurement of right ventricular pressure-volume loops by cardiac catheterization and 2D electrocardiogram. Methods Patients referred for pulmonary hypertension underwent right heart catheterization in our hospital between June 1st, 2015 and June 1st, 2017 are to be enrolled in this study. The right ventricular volume was measured simultaneously by catheter and electrocardiogram. The pressure-volume loops were constructed by the parameters of the pressure and volume in the same cardiac cycle. Results The study completed in four cases and their pressure-volume loops were drawn. The obtained images were irregular and there was no relationship among them. As a result, the construction was a failure. Conclusions The construction of the right ventricular pressure-volume loops of pulmonary hypertension patients by simultaneous catheterization and 2D electrocardiogram is difficult to overcome the technology defects.

BACKGROUNDTo explore the clinical significance of detection of T helper cell (Th1 and Th2) in patients with lung cancer and to provide a foundation for immunological treatment.

METHODSRIA and ELISA were used to detect the level of serum IL-2, IL-4, IL-6, IL-8 and TNF-a in 86 patients with lung cancer, 59 patients with benign pulmonary diseases and 45 healthy people. IL-2 and TNF-a were used to represent cytokines of Th1 type, and IL-4, IL-6 and IL-8 to represent cytokines of Th2 type.

RESULTSThe level of IL-2 [(24.6±12.0) μg/L]in cancer group was significantly lower than that in benign group [(71.1±25.4) μg/L] ( t =3.82, P < 0.01) and normal group [(69.3±19.5) μg/L]( t=2.76, P < 0.01), the level of IL-6 in cancer group [(0.13±0.04) μg/L] was significantly lower than that in normal group [(0.23±0.05) μg/L]( t= 3.39 , P < 0.01), but the levels of IL-4 [(254.2±78.0) μg/L], IL-8 [(0.49±0.16) μg/L], and TNF-a [( 2.76 ±1.12) μg/L] in cancer group were significantly higher than those in benign group [(63.6±18.6) μg/L, ( 0.36 ±0.18) μg/L, (0.96±0.20) μg/L respectively] and those in normal group [(60.9±19.6) μg/L, ( 0.35 ±0.07) μg/L, (0.93±0.19) μg/L respectively] ( t =4.10, 4.89, 3.76 respectively, all P < 0.01). No significant difference of IL-2, IL-4, IL-8 and TNF-a level was observed between benign group and normal group (all P > 0.05). The level of IL-6 in cancer group was similar to that in benign group [(0.15±0.04) μg/L] ( P > 0.05 ). The level of IL-6 in benign group was significantly lower than that in normal group [(0.23±0.05) μg/L] ( P > 0.05 ). There was no significant difference in these cytokines among lung cancer patients with different histological types and in different TNM stages.

CONCLUSIONST helper cell cytokines are out of balance in patients with lung cancer, and this may play a certain role in the pathogenesis of lung cancer. Correcting this immune malfunction may become an important method in lung cancer therapy.

Hepatocellular carcinoma is a common malignant tumor in China, and its global incidence continues to rise and the mortality rate is high. Aberrations in the liver genome lead to malignant transformation of cells and the development of hepatocellular carcinoma, which are also potential therapeutic targets. Different immune cell components in the hepatocellular carcinoma microenvironment, such as tumor-associated macrophages, neutrophils, can promote tumor progression, and cytotoxic T lymphocytes can destroy tumor cells. Different characteristic gene phenotypes in cells can promote or inhibit immune tolerance, which can explain the potential reasons for the sensitivity or resistance of hepatocellular carcinoma patients to immunotherapy, and provide a reference for the exploration of new immunotherapy targets. Further deepening the understanding of the genomic and transcriptomic features in hepatocellular carcinoma and its correlation with immunotherapy can provide new ideas for clinical diagnosis and treatment.

Objective To investigate any protective effect of transplanting EPhrinB2-modified bone marrow mesenchymal stem cells ( BMSCs) with a rat model of cerebral palsy. Methods BMSCs were isolated and cultured, then further modified by lentivirus-mediated transfection of the EPhrinB2 gene. Ninety-six Sprague-Dawley rats were randomly divided into a sham group, a solvent control group ( PBS group) , an empty lentivirus group ( EGFP group) and an EPhrinB2 recombinant lentivirus group ( EPhrinB2 group) , each of 24. A model of cerebral palsy was estab-lished in the rats of the PBS, EGFP and EPhrinB2 groups using hypoxic-ischemic encephalopathy. Seven days after the operation, the lateral ventricles of the PBS, EGFP and EPhrinB2 group mice were injected with phosphate-buff-ered saline solution, BMSCs or EPhrinB2-modified BMSCs respectively. EPhrinB2 protein expression in the hippo-campus was detected using immunohistochemistry 28 days after the operation. The neuron density in the CA1 region of the hippocampus was observed using hematoxylin and eosin staining, and any apoptosis of hippocampal neurons was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling. The expression of nestin and CD31 in the hippocampus was observed using immunofluorescence assays. Morris water maze testing was also conducted to e-valuate changes in learning and memory ability. Results Compared with the other 3 groups, a significant increase in the expression of protein EPhrinB2 was observed in the hippocampuses of the EPhrinB2 group rats. The pathologi-cal changes in the hippocampus among the EPhrinB2 group were significantly less severe than those in the PBS and EGFP groups. The rate of apoptosis in the hippocampuses of the EPhrinB2 group was significantly lower than that of the other groups. Immunofluorescence showed that nestin- and CD31-positive cells were significantly more numerous in the EPhrinB2 group than in the others. In the water maze the average latency of the EPhrinB2 group was signifi-cantly shorter than those of the other groups. Conclusion Lentiviral-mediated EPhrinb2 transfection of BMSCs into the hippocampus can promote EPhrinB2 gene expression, promote angiogenesis and neuron differentiation, inhibit ap-optosis and accelerate the repair of injured nerves.

Objective: To evaluate the pathogenicity of Acinetobacter venetum（Av），which is expected to be used as an environmental remediation agent，using Caenorhabditis elegans（C.elegans）. Methods: The C.elegans were cultured on the media loaded with E.coli OP50 and Av，respectively. The pathogenicity of Av was evaluated by observing the effects of Av on the growth，movement，digestive function，lifespan and reproduction of C.elegans，compared with that of another evaluation system according to NY 1109-2017 General Biosafety Standard for Microbial Fertilizers. Results: By C. elegans system，it was found that the body length，width，head thrash frequency，body bending frequency and average lifespan [（13.5±0.4）d vs.（13.7±0.4）d] of adult nematodes in the Av group were not significantly different from those in the OP50 group（all P>0.05）；while the average time of defecation cycle in the Av group shortened，the total number of progenies in the Av group increased by 18.7%（all P<0.05）. According to NY1109-2017 General Biosafety Standard for Microbial Fertilizers，it was found that the oral LD50 values for both male and female mice were more than 10 g/kgbw，which was practically non-toxic；the pathogenicity test of acute intraperitoneal injection showed that the animals did not have signs of poisoning，deaths or any abnormalities in gross anatomy；Av had no irritation to damaged skin and eyes of rabbits；the hemolysis test was negative；Av was sensitive to seven antibiotics and was medium to one antibiotic. Conclusion Av is not pathogenic. C. elegans can be used in early screening for the pathogenicity of environmental remediation agents.

Objective:To discuss the effect of Fuzheng Ruanjian Anticancer Formula on the malignant biological behaviors of the hepatocellular carcinoma HepG2 cells by requlating protein kinase B(Akt)/murine double minute 2(MDM2)/P53 signaling pathway.Methods:The HepG2 cells were treated with 0,0.05,0.10,0.20,0.40,0.80,1.60,3.20,and 6.40 g·mL-1 Fuzheng Ruanjian Anticancer Formula for 48 h.CCK-8 method was used to detect the survival rates of the HepG2 cells in various groups,and the concentrations of Fuzheng Ruanjian Anticancer Formula for the subsequent experiments were screened.The HepG2 cells were divided into control group,low dose of Fuzheng Ruanjian Anticancer Formula group(0.2 g·mL-1),medium dose of Fuzheng Ruanjian Anticancer Formula group(0.4 g·mL-1),high dose of Fuzheng Ruanjian Anticancer Formula group(0.8 g·mL-1),SC79 group(8 mg·L-1 SC79),and high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group(0.8 g·mL-1 Fuzheng Ruijian Anticancer Formula+8 mg·L-1 SC79).CCK-8 method was used to detect the proliferation activities of the HepG2 cells in various groups;clone formation assay was used to detect the clone formation rates of the HepG2 cells in various groups;flow cytometry was used to detect the apoptotic rates of the HepG2 cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion HepG2 cells in various groups;Western blotting method was used to detect the expression levels of proliferating cell nuclear antigen(PCNA),cysteine aspartate specific proteinase(Caspase-3),matrix metalloproteinase(MMP)-2,MMP-9,phosphorylated Akt(p-Akt),phosphorylated MDM2(p-MDM2),and P53 proteins in the HepG2 cells in various groups.Results:As the increasing of concentrations of Fuzheng Ruanjian Anticancer Formula(0,0.05,0.10,0.20,0.40,0.80,1.60,3.20,and 6.40 g·mL-1),the surival rates of the HepG2 cells were gradually decreased(P<0.05),and 0.2,0.4,and 0.8 g·mL-1 Fuzheng Ruanjian Anticancer Formula were selected for the subsequent experiments.The CCK-8 assay results showed that compared with control group,the proliferation activities of the HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05),in a dose-dependent manner,while the proliferation activity of the cells in SC79 group was significantly increased(P<0.05).Compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the proliferation activity of the HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased(P<0.05).The clone formation assay results showed that compared with control group,the clone formation rates of the HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the clone formation rate of the cells in SC79 group was significantly increased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the clone formation rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased(P<0.05).The flow cytometry results showed that compared with control group,the apoptotic rates of the HepG2 cells in low,medium,and high doses of Fuzheng Ruijian Anticancer Formula groups were significantly increased(P<0.05)in a dose-dependent manner,while the apoptotic rate of the cells in SC79 group was significantly decreased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the apoptotic rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly decreased(P<0.05).The Transwell chamber assay results showed that compared with control group,the numbers of migration and invasion HepG2 cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the numbers of migration and invasion cells in SC79 group were significantly increased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the numbers of migration and invasion HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased(P<0.05).The Western blotting results showed that compared with control group,the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in low,medium,and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased(P<0.05)in a dose-dependent manner,while the expression levels of Caspase-3 and P53 proteins were significantly increased(P<0.05)in a dose-dependent manner,while the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in SC79 group were significantly increased(P<0.05),and the expression levels of Caspase-3 and P53 proteins were significantly decreased(P<0.05);compared with high dose of Fuzheng Ruanjian Anticancer Formula group,the expression levels of PCNA,MMP-2,MMP-9,p-Akt,and p-MDM2 proteins in the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased(P<0.05),while the expression levels of Caspase-3 and P53 proteins were significantly decreased(P<0.05).Conclusion:Fuzheng Ruanjian Anticancer Formula may inhibit the proliferation,migration,and invasion of the HepG2 cells and promote the apoptosis,and its mechanism may be related to suppressing the Akt/MDM2 signaling pathway and upregulating the P53 proteim expression.

Objective To study the clinical characteristics of immune tolerance after liver transplantation in children and to identify possible predictors.Methods The clinical data of 37 pediatric patients who underwent liver transplantation between April 2006 and April 2014 at the Children's Hospital of Chongqing Medical University were retrospectively analyzed.The patients were divided into the no-drug (n =4),single-drug (n =16) and multi-drug (n =17) groups according to the status of their current immunosuppressant medications.The possible predictive factors were screened based on their clinical data,and statistical analysis was performed.Results The 37 liver transplantation recipients included 16 males (43.2％) and 21 females (56.8％).The factors that differed among the groups included age at transplantation and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of the transplant recipients.Age,ALT level,and AST level of the transplant recipients were significantly different between the single-drug group and the multi-drug group (all P ＜ 0.05).However,only the ALT Ievel was significantly different (P ＜ 0.05) between the no-drug group and the multi-drug group.No significant differences were found in the various other factors between the no-drug and single-drug groups.Conclusion The age of the recipient at transplantation was a predictive factor affecting clinical immune tolerance in pediatric liver transplantation,while ALT and AST levels were potential predictors of postoperative immune tolerance.