Objective To evaluate the effect of NGX6 combined with cisplatin on the inhibition rate of A549 cells and NCI-H1975 cells and antitumor effects in vivo.Methods The NGX6 was loaded in the LPD, and prepare Liposome protamine DNA complexes.A549 cells and NCI-H1975 cells were seperately divided into NGX6 group ( 30 μg/mL NGX6 concentration ) , cisplatin group, NGX6 +cisplatin group, PBS as negative control group.The effect of cytotoxicity of four group on A549 cells and NCI-H1975 cell were evaluated by MTT assay.the clone forming rate and the inhibition rate were determined by Cell colony count.lung transplantation tumor model were successfully established, then nude mice were divided into four groups as abeve, each group of 10, tumor size and survival period were determined tumor cell apoptosis were observed.Results The cell viability of A549 cells and NCI-H1975 cells of (NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The cloning efficiency of A549 cells and NCI-H1975 cells of ( NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The tumor inhibitory rate was in vivo for (NGX6 +cisplatin) was higher than other group(P<0.01).The median survival of nude mice in (NGX6 +cisplatin), NGX6, cisplatin and saline group were 43,31,29 and 15 days.Conclusion NGX6 combination with cisplatin can inhibit the cell proliferate of lung cancer cells and inhibit the tumor growth and the combination of NGX6 and cisplatin may be a potentially effective treatment for lung cancer.

Objective To observe the interfering effect of different doses of penehyclidine hydrochloride (PHC) on the mRNA expressions of nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) in the lung tissue of rats with traumatic shock so as to investigate the protective role of PHC in secondary long injury following traumatic shock and the underlying mechanism.Methods The traumatic shock model was established.A total of 104 Wistar rats were randomly divided into four groups:control group,shock group,low dose PHC group ( P1 group) and high dose PHC group ( P2 group).At the beginning of resuscitation,the rats in P1 and P2 groups were given transjugular intravenous injection of 2 ml/kg isotonic saline containing 0.15 mg/kg and 0- 45 mg/kg PHC respectively,while the rats in shock and control groups were injected only isometric isotonic saline.The rats in the four groups were killed at 2 h,6 h,12 h and 24 h after resuscitation respectively to detect the mRNA expressions of NF-κB and iNOS by using RT-PCR and determine the lung wet/dry weight (W/D) ratio,lung permeability index (LPI) and lung injury score (LIS).Results The mRNA expressions of NF-κB and iNOS,lung W/D ratio,LPI and LIS at all the time intervals in the shock,P1 and P2 groups were all significantly increased as compared with those in the control group (P＜0.05).Howerver,the P2 group showed significant reduction in aspects of the mRNA expressions of NF- κB and iNOS,lung W/D ratio,LPI and LIS at all time points and P1 group also had significant decrease regarding the mRNA expressions of NF-κB and iNOS,lung W/D ratio at2 h,6 h,and LPI and LIS at 2 h,6 h,12 h,as compared with the shock group.Meanwhile,P2 group showed evident decrease at 6 h concerning the mRNA expressions of NF-κB and iNOS,lung W/D ratio,LPI and LIS as compared with P1 group (P ＜ 0.05 ).Conclusions PHC,especially at a large dosage,can significantly mitigate the long injury secondary to traumatic shock,and the mechanism may be associated with the inhibition of mRNA expressions of NF-κB and iNOS.

OBJECTIVETo investigate the effect of pulsed electromagnetic fields (PEMs) on inducing osteoclastic like cell (OLC) formation changes and apoptosis in ovariectomized (OVX) rats bone marrow culture in vitro.

METHODSThirty healthy three-month-old female Wistar rats were either sham-operated (Sham) or ovariectomized (OVX) and randomly divided into three groups: group A (OVX + PEMs, 18 rats), group B (OVX, 6 rats) and group C (Sham, 6 rats); group A was again randomly divided into three groups: A1, A2, A3. The frequencies adopted were 1.5, 2, 75 Hz and 30 minutes for once a day. All rats were fed with normal diet for 3 months, then the bone marrow of all rats were cultured, 2 days later, group A cells (including group A1, A2, A3) were collected and exposed to different frequencies PEMs for 2 weeks (30 min/day). In order to observe the changes of osteoclasts and count their numbers, cells were taken for Wright Giemsa staining, tartrate-resistance acid phosphatase (TRAP) staining and Hoechst 33258 staining.

RESULTSTRAP staining results indicated the number of OLC in group C was the least, then was group A2, A3, A1, B. The number of OLC in group B was remarkably increased (P < 0.01; vs group C, A2). The number of OLC in group B was significantly increased (P < 0.05; vs group A1, A3). Hoechst 33258 staining results indicated the number of apoptosis of OLC in group C was more than other groups, which of group C, A2 was significantly increased (P < 0.05; vs group B).

CONCLUSIONPEMs had decreased the formation of OLC and increased the number of apoptosis of OLC in ovariectomized (OVX) rats bone marrow culture in vitro, the effects of 2 Hz was the best. PEMs would be a new way of osteoporosis therapy.

Acid Phosphatase ; metabolism ; Animals ; Apoptosis ; radiation effects ; Bone Marrow Cells ; cytology ; enzymology ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Female ; Isoenzymes ; metabolism ; Osteoclasts ; cytology ; enzymology ; radiation effects ; Ovariectomy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase

Objective To investigate the level of cadmium(Cd)in commercial aquatic products in Xiacheng District, Hangzhou. Methods We randomly collected 293 aquatic products which belonged to six aquatic animals in the markets in Xiacheng District to determine the content of Cd. It was further evaluated by single factor pollution index(PI)according to the standard GB 2762-2017. In 11 samples of swimming crabs, Cd was examined in the different parts. Results There was no significant difference in the content of Cd between the samples collected in the markets and those in the supermarkets. It significantly differed in the samples of different aquatic animals(P < 0.05). The prevalence of Cd that exceeded the standard was as follows: seawater crustaceans(28.6%) > cephalopods(11.1%) > freshwater crustaceans(8.4%) > bivalves(6.9%). However, it was not excessive in the samples of fish. The mean level of Cd in the seawater crustaceans was 0.466 3 mg/kg, which resulted in the proportion of the samples that were excessive being 28.6%. Particularly, the mean level in sea crab was as high as 1.101 mg/kg with the proportion being 66.7%. In the samples of swimming crabs, there was a significant difference in the prevalence of Cd between swimming crab gonads and crab chests or legs(P > 0.05), while no statistical difference between crab chests and legs(P > 0.05). Conclusion The content of Cd in the aquatic products may be excessive in Xiacheng District, which warrants additional regulatory efforts to food safety. The public should reduce the consumption of aquatic products with high content of Cd.

OBJECTIVEUsing molecular epidemiology method to characterize human immunodeficiency virus type 1 (HIV-1) subtype CRF01 _ AE strains being prevailed in Zhejiang province.

METHODSGag fragments of the HIV-1 strains were amplified by nested-polymerase chain reaction (nPCR) from the DNA extracted from whole blood of HIV-1 infected individuals in Zhejiang province. PCR products were sequenced and analyzed by phylogenetic method.

RESULTS81 HIV-1 subtype CRF01 _ AE sequences were identified from the 192 samples that sequenced successfully. As one of the dominant subtypes in Zhejiang, CRF01 _ AE was transmitted mainly by heterosexual or homosexual contact in local residents. In migrants living in Zhejiang, CRF01 _ AE were transmitted mainly by heterosexual contact or injecting drug use. There were three main clusters in the phylogenetic tree which bootstrap value was larger than 60. We named the clusters with group MIX (47 sequences), group SEX (7 sequences) and group MSM (12 sequences) based on the transmission. Pairwise DNA distances in the gag region within the three groups and between CM240 were different (P = 0.000). Data through the analyses of deduced amino acid sequences from the three groups showed that several signature amino acid sites were distinct from the same positions of the subtype reference strains.

CONCLUSIONThe CRF01 _ AE strain prevailing in Zhejiang province was from several sources, transmitted by more than three different transmission routes, and becoming the main subtypes circulating in homosexual population in this study. More attention needs to be paid to the epidemic characteristic of CRF01 _ AE.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; methods ; Phylogeny ; Young Adult

To develop polyclonal antibodies against predicted B cell epitopes in HIV-1 accessory protein Vpr, the prepared consensus Vpr amino acid sequence was used to predict potential B cell epitopes by online softwares (ABCpred and Bcepred), the synthesized polypeptides of B-cell epitopes were subsequently conjugated with keyhole limpet hemocyanin (KLH) and then used to immunize rabbits. The antibody titers were determined by ELISA, and antibody specifity was analyzed by Western-Blotting and immunoprecipitation, respectively. Amino acid residues 3-19 (N) and 82-95 (C) of Vpr were predicted as the potential B cell epitopes. After inoculation of the conjugation of synthesized peptide to KLH, the antibody titers in rabbit sera against N and C peptides reached more than 1:100000 by ELISA. Western-Blotting analysis showed that the polyclonal antibodies reacted with both wild Vpr and fusion protein of GFP with Vpr, no matter Vpr was derived from HIV-1 B subtype or CRF07_BC recombinant form; Immunoprecipitation analysis showed similar reactions to Western-Blotting results. Two B cell epitopes of Vpr were successfully predicted by Bio-informatics methods and polyclonal antibodies against those peptides could be successfully prepared.
Animals ; Antibodies, Viral ; blood ; immunology ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; HIV Infections ; blood ; immunology ; virology ; HIV-1 ; genetics ; immunology ; Humans ; Peptides ; genetics ; immunology ; Rabbits ; vpr Gene Products, Human Immunodeficiency Virus ; genetics ; immunology

To investigate the effects of HIV-1 infection on the expression of host factors TSG101 (Tumor Susceptibility Gene 101) and Alix (ALG-2-interacting protein X). HIV-1 infectious clone pNL4-3 was used to infect TZM-bl, PM1, Jurkat cell lines and human peripheral blood mononuclear cells (PBMC). Twenty-four hours post-infection, the infected or uninfected cells were harvested respectively for extraction of total RNAs and total cellular proteins, which were subsequently used in RT-PCR and Western-blotting respectively to quantify TSG101 and Alix, respectively. Our data showed that HIV-1 infection resulted in various influences on the expression of TSG101 and Alix in the cell lines and the primary PBMC. A down-regulation was mainly observed in the cell lines, whereas an up-regulation of TSG101 was identified in primary PBMC. Three patterns were observed for down-regulation, including dual down-regulation of TSG101 and Alix for Jurkat cells, single down-regulation of Alix for TZM-bl cells and marginal or no influence on PM1 cells. The dual down-regulation of Alix and TSG101 in Jurkat cells coincided with less expression of HIV-1 p24 protein. This is the first-line evidence that HIV-1 infection affects the expression of host factors TSG101 and Alix, the down-regulation of these molecules may influence the HIV-1 replication. The underlying mechanism remains to be addressed.
Calcium-Binding Proteins ; genetics ; metabolism ; Cell Cycle Proteins ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Endosomal Sorting Complexes Required for Transport ; genetics ; metabolism ; Gene Expression Regulation ; HEK293 Cells ; HIV-1 ; physiology ; Humans ; Jurkat Cells ; Leukocytes, Mononuclear ; metabolism ; virology ; RNA, Messenger ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

BACKGROUNDThe CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.

METHODSA SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.

RESULTSOne SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.

CONCLUSIONSWe conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.

Animals ; Chimera ; Genes, env ; HIV-1 ; genetics ; physiology ; Humans ; Macaca mulatta ; Proviruses ; genetics ; Receptors, CCR5 ; physiology ; Simian Immunodeficiency Virus ; genetics ; physiology

OBJECTIVETo study the specific amino acid variation in Nef that may be related to disease progression after infection with HIV-1 subtype B, a predominant strain circulating in China, and to determine whether changes in Nef secondary structure may influence different stages of AIDS development based on the concept that the Nef gene of HIV infection dramatically alter the severity of viral infection and virus replication and disease progression, and that long-term non-progressors (LTNP) of HIV infection are commonly associated with either a deletion of the Nef gene or the defective Nef alleles.

METHODSThe study subjects were divided into LTNP1(n=14), LTNP2 (n=16) and slow progressor (SP, n=19) groups for mutational analysis of the Nef sequence. The data were obtained by using Bioedit, MEGA, Anthewin and SAS software.

RESULTSResidues in Nef TA(48/49) and K151 occurred more frequently in the LTNP group while AA(48/49) was more frequently observed in the SP group. Of the differences observed in the secondary structure comparison using Nef consensus sequences of these three groups, one was roughly corresponding to the Nef(48/49) mutation site.

CONCLUSIONTA(48/49), K(151), and AA(48/49) in the Nef gene might be associated with the different stages of HIV infection, and there may be a link between the Nef secondary structure and the progression of HIV-1 infection.

Amino Acid Sequence ; Base Sequence ; Blood Donors ; CD4 Lymphocyte Count ; China ; epidemiology ; Disease Progression ; Gene Products, nef ; genetics ; HIV Infections ; epidemiology ; virology ; HIV Long-Term Survivors ; HIV-1 ; classification ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; genetics ; Time Factors

The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.
Animals ; Animals, Domestic/blood/microbiology/parasitology ; Antibodies, Fungal/*blood ; Antibodies, Protozoan/*blood ; China/epidemiology ; Encephalitozoon cuniculi/*immunology/isolation & purification ; Encephalitozoonosis/blood/microbiology/*veterinary ; Female ; Male ; Rabbits/blood/microbiology/parasitology ; Seroepidemiologic Studies ; Toxoplasma/*immunology/isolation & purification ; Toxoplasmosis, Animal/*blood/parasitology