Aim To investigate the enhanced efficacy and reduced toxicity of GanoExtra in combination with cyclophosphamide(CTX)on inhibiting lung metastasis and immune function in mice.Methods The CCK-8 method was used to verify the cytotoxic effects of Gano-Extra on MCF-7 and 4T1 tumor cells.In vivo experi-ment,a mouse model of lung metastasis of breast canc-er was established by injecting 4T1 tumor cells into the tail vein,which was divided into four groups including 4T1 model group,CTX group,GanoExtra group and GanoExtra+CTX group.The control group was set.After 21 days,the mice were euthanized under anes-thesia,and the body weight of the mice was recorded.Average lung index and spleen index were calculated.The mouse spleen lymphocyte transformation experi-ment was used to determine the activity of spleen cells.The NK cell activity assay was used to determine the activity of NK cells.Blood cells were determined in mouse blood samples.Flow cytometry was used to de-termine the levels of CD4+and CD8+T cells in blood samples.ELISA was used to measure the levels of TNF-α and IL-6 in serum.HE staining was used to ob-serve the pathological morphological changes in tumors and various tissues;and CFSE staining was used to de-termine the proliferative effect of GanoExtra on CD8+cells.Results In vitro GanoExtra at 50 mg·L-1 sig-nificantly inhibited the activity of MCF-7 and 4T1 tumor cells.In the breast cancer pulmonary metastasis model,compared with the model group,the spleen in-dex and blood WBCs content were significantly re-duced,while the activity of NK cells,spleen cells,and the proportion of RBCs,CD 3+and CD 8+T cells in the blood were significantly increased.At the end of the treatment,compared with the CTX group,the number of lung metastases and lung index of the Gano-Extra+CTX group were significantly reduced,and the levels of HGB,CD8+cells,IL-6,and TNF-α in the blood of mice were significantly increased.GanoExtra at 10 mg·L-1 significantly promoted the proliferation of CD8+T cells in vitro.Conclusions GanoExtra can enhance the inhibitory effect of CTX on tumor metasta-sis,alleviate adverse reactions such as splenomegaly and pulmonary enlargement caused by CTX,and have a health-promoting function of promoting the prolifera-tion of CD8+T cells to enhance the immune efficacy of the body.

Objective:To investigate the application value of single-sperm sequencing in resolving the carrier status of preim-plantation genetic testing(PGT)for chromosomal structural rearrangements in Robertsonian translocations.Methods:Haplotypes were constructed by single-sperm isolation combined with single-sperm sequencing for a patient with 45,XY,der(13;14)(q10;q10).Twenty single-sperm samples were isolated by mechanical braking and subjected to whole-genome amplification(WGA),and then the Asian Screening Array(ASA)gene chip was used to detect the 183 708 single nucleotide polymorphisms(SNP)of the WGA products.The single sperm associated with the translocation that could be used as haplotype inference was detected by copy number variation(CNV)sequencing,and the chromosomal haplotypes with normal and Robertsonian translocations were inferred.Three biopsy samples of embryonic trophoblast cells were used as the objects.After whole-genome amplification,high-throughput sequencing was employed to determine the status of the translocation chromosome carried by the embryos.The available blastocysts were selected for transfer,and the amniotic fluid samples were taken at 18 weeks of gestation to confirm whether the fetus carried the pathogenic muta-tion.Results:A total of 6 037 SNP sites were screened by single-sperm sequencing,and 30 sites selected to distinguish normal and translocation haplotypes.Preimplantation haplotype analysis showed that all the three embryos were euploids without Robertsonian translocation chromosome.Genetic testing of amniotic fluid in the second trimester confirmed that the karyotype of the fetus was 46,XN,carrying no Robertsonian translocation chromosome.Conclusion:For male carriers of Robertsonian translocation,single sperm sequencing can be used to screen SNP sites to construct haplotypes for distinguishing normal and Robertsonian translocation em-bryos,and to provide a basis for embryo selection by preimplantation chromosomal structural genetic testing.

Tanreqing injection is suitable for early pneumonia, acute bronchitis, acute exacerbations of chronic, and upper respiratory tract infection which are classified with phlegm-heat obstructing lung syndrome of traditional Chinese medicine. To understand the clinical adaptation syndromes and medication characteristics of the post-market Tanreqing injection, the research team of the paper monitored the patients who are used with Tanreqing injection from September 2012 to October 2013 in four leader hospitals based on the method--prospective, multi-center, large sample, registration-type hospital centralized monitoring,and analyzes the general information, diagnostic information and medication characteristics of patients, in order to produce evidence for clinical practice and medication decisions and to establish the foundation of rational drug use.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; therapeutic use ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Prospective Studies ; Respiratory Tract Infections ; Young Adult

The paper is focused on the clinical applications of Tanreqing injection after listing, detecting and analyzing the related blood indicators of patients with allergic reactions based on prospective, multi-center, large sample, registration-type clinical safety monitoring nested case-control study (NCCS) to explore the possible mechanisms of allergic reaction of Tanreqing injection, 3 006 patients cases used with Tanreqing injection were monitored, including 3 cases of adverse reactions and 2 cases of allergic reactions. Each patient of allergic reactions, according to the ratio of 1:4 matches four cases of not adverse reactions as a control group of patients, while 5 healthy and 5 cases of volunteers into the healthy group. We examined the correlation detection of cases of allergic reactions among groups such as T-IgE, IgA, IgG, C3, C4, IFN-g, IL-2, IL-4, IL-6, IL-10. Allergic reactions of Tanreqing injection may be mediated by IgE as type I based on existing research data. This results and conclusions will promote the justifiability and safety of clinical applications.
Adolescent ; Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Hypersensitivity ; diagnosis ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Male ; Prospective Studies ; Young Adult

Objective To study the effects of living environment conditions and animal-animal interaction on the blood hormone levels and psychological behavior in Chinese tree shrews .Methods Chinese tree shrews were raised in ca-ges of different space sizes or were administered reserpine for 15, 30, 60, 120, 180 days, respectively.Then the animals were anesthetized by ether inhalation , and blood samples were taken from the heart to detect the levels of blood testosterone (T), estradiol (E2), endothelin (ET), adrenaline (A) and noradrenaline (NA) by radioimmunoassay (RIA).Results 1.Chinese tree shrews were bred in large cage (D1group) or small cage (X1 group) for 15, 30, 60, 120, 180 days, re-spectively.Compared with the animals bred in the large cage (D1 group), the level of blood testosterone (T) was signifi-cantly reduced (P<0.01), and the levels of adrenaline , noradrenaline and endothelin were significantly increased in the small cage group (P<0.01 for all).2.The animals raised in small cages in close neighborhood with large cages for 15, 30, 60, 120, 180 days, respectively.The levels of testosterone, adrenaline and noradrenaline in the large cage group (X2 group) were significantly higher than those of the small cage group (X1 group) (P<0.01 for all).3.The adrenaline and noradrenaline levels were significantly lower in all the reserpine groups (P<0.01 for all).4.The animals bred in small cages (X1 group) and in small cages in close neighborhood with large cages showed sudden loss , reduced appetite, testicu-lar atrophy , penile prolapse and stress symptoms .The animals of the reserpine groups appeared gentle temperament , signif-icantly reduced activity and reduced appetite .However , after stopping the reserpine administration and feeding them in large cages , the animals gradually returned to normal behavior .Conclusion Both animals living environment conditions and animal-animal interaction may cause changes of blood hormone levels and psychological behavior in Chinese tree shrews.

BACKGROUNDData on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.

METHODSThe survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.

RESULTSThe analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).

CONCLUSIONSThe prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.

Adult ; Aged ; Awareness ; Female ; Humans ; Hypertension ; complications ; epidemiology ; therapy ; Male ; Middle Aged ; Prevalence ; Renal Insufficiency, Chronic ; complications

It is of vital significance to conduct active post-marketing surveillance of Chinese medicine, as an active response to laws, rules and guidelines issued by the China food and drug administration. The standards for technological specifications based on expert consensus have been drafted. These will provide technological support in evaluating adverse drug reactions (ADRs) or adverse drug events (ADEs). The technological specifications for post-marketing surveillance focus on two surveillance designs; one is a large sample registry study to explore general population ADR/ADE characteristics, the other is a nested case-control study to explore the characteristic and mechanisms of ADRs.
China ; epidemiology ; Drug Monitoring ; adverse effects ; standards ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; adverse effects ; standards ; Hospital Information Systems ; Humans ; Product Surveillance, Postmarketing ; methods ; standards

Osteosarcoma is the most common primary malignant bone cancer in children and adolescents. Emerging evidence has suggested that the capability of a tumor to grow is driven by a small subset of cells within a tumor, termed cancer stem cells (CSCs). Although several methods have been explored to identify or enrich CSCs in osteosarcoma, these methods sometimes seem impractical, and chemotherapy enrichment for CSCs in osteosarcoma is rarely investigated. In the present study, we found that short exposure to chemotherapy could change the morphology of osteosarcoma cells and increase sarcosphere formation in vitro, as well as increase tumor formation in vivo. Furthermore, methotrexate (MTX)-resistant U2OS/MTX300 osteosarcoma cells were larger in size and grew much more tightly than parental U2OS cells. More importantly, U2OS/MTX300 cells possessed a higher potential to generate sarcospheres in serum-free conditions compared to parental U2OS cells. Also, U2OS/MTX300 cells exhibited the side population (SP) phenotype and expressed CSC surface markers CD117 and Stro-1. Notably, U2OS/MTX300 cells showed a substantially higher tumorigenicity in nude mice relative to U2OS cells. Therefore, we conclude that chemotherapy enrichment is a feasible and practical way to enrich osteosarcoma stem cells.
Animals ; Antigens, Surface ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; drug effects ; pathology ; Osteosarcoma ; metabolism ; pathology ; Phenotype ; Proto-Oncogene Proteins c-kit ; metabolism

Objective The proto-oncogene c-Met was found to express on human laryngeal carcinoma Hep-2 cell line in previous research. In the present study, the author further examined whether inhibition of c-Met by RNA interference(RNAi) might inhibit biologic activity of Hep-2 cell line in vitro and proliferation using a murine laryngeal carcinoma model. Methods RNAi plasmid that can express small interfering RNA targeting c-Met or siRNA that did not match any known human coding mRNA( control siRNA plasmid) was designed,constructed,and transfected into Hep-2 cell line by using cationic liposome Lipofectamine2000 as transfecting agent. In vitro, the transfection efficacy was tested by RT-PCR and Western Blot method, then elected the most inhibitive c-Met-siRNA sequence. Cell proliferation, movement and invasion were studied using MTT, cell migration assay and cell invasion assay, respectively. The Hep-2 cells were transplanted into nude mice, then the time of tumor formation and growth were observed. After tumor formation, c-Met-siRNA was given as the anti-tumor therapy. Expression of c-Met, MMP-9 and VEGF were detected by Western Blot method. Results After the pSilencer2. 0/c-Met-shRNA recombinant plasmid transfeetion into laryngeal carcinoma Hep-2 cells, the expression of mRNA and proteinum of c-Met decreased significantly in Hep-2 cells. On the 35th day after tumor vaccination, the tumor volume was (138±27)mm~3 in c-Met-siRNA transfection group, Which was diminished significantly in contrast with control group( P<0. 01 ). The expression of c-Met, MMP-9 and VEGF in the tumor of experiment group was decreased significantly, respectively (P<0.05 ). Conclusions The results indicated that c-Met-siRNA can down-regulate the expression of c-Met and markedly inhibit laryngeal carcinoma Hep-2 cell proliferation, movement and invasion and the growth of transplantation tumor of nude mice. The siRNA expressing plasmid mediated gene therapy might be a new strategy in targeting molecular therapy of cancer of larynx.

OBJECTIVETo compare the therapeutic effect of recombinant human brain natriuretic peptide (rhBNP) and nitroglycerin on acute decompensated heart failure (ADHF).

METHODSFifty ADHF patients were randomly divided into rhBNP group and nitroglycerin group. In all the patients, dyspnea and global clinical status were assessed before and at 30 min, 6 h and 24 h after drug administration, and the volume of fluid intake and urine along with hemodynamic parameters was recorded 24 h after drug administration. In the nitroglycerin group, the patients received an initial nitroglycerin dose of 5 microg/min, with subsequent dose increment of 5 microg/min every 3 to 5 min; the dose was adjusted individually according to the hemodynamics of the patients. The patients in rhBNP group were given rhBNP at the initial dose of 1.5 microg/kg by with an intravenous bolus injection followed by infusion at the rate of 0.0075 microg.kg(-1).min(-1) for 72 h.

RESULTSAt 30 min and 6 h after drug administration, the patients in the rhBNP group showed significant greater improvement of dyspnea (P=0.042 and 0.019) and global clinical status (P=0.018 and 0.044) than those in the nitroglycerin group, but 24 h after drug administration, no significant difference was noted between the two groups (P=0.192 and 0.179). Twenty-four hours after drug administration, the mean urine volume was significantly greater in rhBNP group than in nitroglycerin group (1513.8-/+242.9 vs 1341.2-/+239.7 ml, P=0.015), and the ejection fraction increased and pulmonary arterial pressure and systolic blood pressure decreased at greater amplitude in the former group (P=0.001,0.000 and 0.002, respectively). At 72 h, the numbers of premature ventricular contraction and couplets premature beats and onset of paroxysmal ventricular tachycardia were significantly reduced in rhBNP group as compared with the nitroglycerin group (P=0, 0.001 and 0.002, respectively).

CONCLUSIONRhBNP promotes urine excretion, decreases pulmonary arterial pressure and increases left ventricular ejection fraction to improve dyspnea and global clinical status and reduce the onset of ventricular arrhythmia in ADHF patients.

Aged ; Aged, 80 and over ; Blood Pressure ; drug effects ; Female ; Heart Failure ; drug therapy ; pathology ; physiopathology ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; administration & dosage ; genetics ; therapeutic use ; Nitric Oxide Donors ; administration & dosage ; therapeutic use ; Nitroglycerin ; administration & dosage ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Treatment Outcome