OBJECTIVETo investigate the distribution and proportion of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Autonomous Region.

METHODS152 HIV-1 patients were enrolled from 11 cities in Guangxi Autonomous Region from 2010 to 2012 by convenient sampling. Inclusion criterias were listed as the fdlowing: HIV-1 infection was confirmed by Western blot, HIV-1 viral load >1 000 copies/ml, > 18 year-old, and without any serious illnesses. 5 ml of peripheral blood samples were obtained from each patient. The viral RNA was isolated from plasma and used for amplification of full-length pol gene by nested RT-PCR. The amplified products were sequenced. After editing and modification, all sequences were characterized for preliminary subtyping by genotyping and confirmed with phylogenetic tree constructed by MEGA 5.03 software. The recombinant identification of 2 unknown recombinant strains was determined by RIP and jpHMM at GOBICS.

RESULTSAmong 152 patients, 137 full-length pol genes were successfully amplified and 127 HIV-1 subtypes were identified. The distribution and proportion of subtypes was summarized as the following 71 cases of CRF01_AE, accounting for 55.9% (71/127), 38 CRF08_BC, 29.9% (38/127), 13 CRF07_BC, 10.2% (13/127), and 3 B (B'), 2.4% (3/127), 2 unknown recombinant strains, 1.6% (2/127). In 11 cites of Guangxi Autonomous Region, subtype CRF01_AE was the dominant strain. Among heterosexual transmitted patients and drug abusers, the proportions of subtype CRF01_AE were 67.4% (58/86) and 34.1% (14/41), respectively. There was a significance different in the distribution of CRF01_AE in different routes of transmission (χ(2)=15.07, P<0.001). In age 21- 35, age 36- 60 and age>60 groups, the proportions of CRF01_AE was 43.6% (17/39), 57.6% (38/66), 77.3% (17/22), and CRF08_BC was 43.6% (17/39), 28.8% (19/66), 9.1% (2/22), respectively, the difference in proportions was significant(χ(2)=8.48, P= 0.014). The patterns of two unknown recombinant strains were found to be CRF01_AE/B (B') and CRF01_AE/C/B(B'), respectively.

CONCLUSIONCRF01_AE was the dominant HIV-1 subtype in Guangxi Autonomous Region from 2010 to 2012, with heterosexual transmission as its main spreading route. The two unknown recombinant strains in Guangxi Autonomous Region were reconstructed by subtype CRF01_AE and CRF_BC.

Blotting, Western ; China ; epidemiology ; Cities ; Drug Users ; Genes, pol ; Genotype ; HIV Infections ; epidemiology ; transmission ; virology ; HIV-1 ; genetics ; Humans ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; blood ; pol Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo generate a mouse model of chronic hepatitis B (CHB) infection by performing in vivo transduction of hepatitis B virus (HBV) covalently closed circular (ccc)DNA.

METHODSNude mice were injected with HBV cccDNA at doses of 1.5, 1.0 or 0.5 mug/ml. A control group was generated by giving equal injection volumes of physiological saline. The serum levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) on post-injection days 1 and 3, weeks 1-6, 8 and 10 were assayed by reflection immunoassay. At post-injection week 10, all animals were sacrificed and liver tissues were collected. Copies of HBV DNA in serum and liver tissue were detected by real-time PCR. HBV antigens in liver tissue were detected of by immunohistochemistry. Pathological analysis of liver tissue carried out with hematoxylin-eosin staining. Linear correlation of data was determined by statistical analysis.

RESULTSHBsAg and HBeAg were detected in sera from all three groups of cccDNA-injected mice staring at post-injection day 1 and lasting through week 10. The levels of HBsAg over the 10-week period showed two patterns of increase-decrease;the lowest level was detected at week 4 and the highest level was detected at week 8. In contrast, the levels of HBeAg over the 10-week period showed three patterns of increase-decrease; the lower levels were detected at weeks 2 and 4 and the higher levels at weeks 3 and 6. HBV DNA copies in liver tissues showed a cccDNA dose-dependent descending trend over the 10-week study period (1.5 mug/ml:1.14E+07 ± 6.51E+06 copies/g, 1.0 mug/ml:9.81E+06 ± 9.32E+06 copies/g, and 0.5 mug/ml:3.72E+06 ± 2.35E+06 copies/g; Pearson's r =0.979). HBV DNA copies in sera showed the pattern of 1.0 mug/ml cccDNA more than 1.5 mug/ml cccDNA more than 0.5 mug/ml cccDNA, and in general were higher than those detected in the liver tissues. Liver tissues from all cccDNA-injected mice showed positive immunohistochemistry staining for both HBsAg and HBeAg. HE staining showed that the liver tissues of all cccDNA-injected mice had severe fatty and vacuolar degeneration and less obvious structure of liver lobules (compared to the liver tissues from control mice).

CONCLUSIONThe CHB mouse model successfully established in this study by in vivo transduction of HBV cccDNA may represent a useful tool to study the pathogenic mechanisms and potential antiviral treatments of human CHB.

Animals ; DNA, Circular ; administration & dosage ; DNA, Viral ; administration & dosage ; Disease Models, Animal ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; virology ; Male ; Mice ; Mice, Nude ; Transduction, Genetic ; Virus Replication

OBJECTIVETo prepare a pH fluorescence probe based on styrylcyanine dyes for live cell imaging.

METHODSThe Probe 1 was prepared by reaction of 4-pyridinecarboxaldehyde with 1,1,2-trimethylbenz[e]indole. The influence of pH on the fluorescent properties was examined, and the cell viability was examined using cell counting kit-8. The Probe 1 was used as a pH fluorescence probe in living cell.

RESULTSProbe 1 emitted green fluorescence under neutral and basic conditions but orange fluorescence under acid condition. Probe 1 selectively stained the cytoplasmic regions of living cells without significantly affecting the cell viability.

CONCLUSIONThe pH-sensitive fluorescent probe prepared based on styrylcyanine possesses good ability of cell membrane permeation for live cell fluorescent imaging.

Cells, Cultured ; Fluorescence ; Fluorescent Dyes ; Hydrogen-Ion Concentration ; Optical Imaging

BACKGROUNDChemotherapy is very important in treatment of advanced non-small cell lung cancer (NSCLC), and the third-generation cisplatin-based chemotherapy regimens have been the standard treatment for advanced NSCLC. The aim of this study is to compare the efficacy and toxicity among four different chemotherapeutic regimens combined with radiotherapy in patients with stage III/IV NSCLC.

METHODSA total of 527 patients with stage III/IV NSCLC were enrolled, among whom there were 243 patients received cisplatin/vinorelbine (NP group), 163 patients for cisplatin/paclitaxel (TP group), 65 patients for cisplatin/gemcitabine (GP group) and 56 patients for cisplatin/docetaxel (DP group). The efficacy, side effects, median time to progression (TTP), median survival time (MST), 1- and 2-year survival rate were compared.

RESULTSThe response rate was 46.9% in the NP arm, 44.8% in the TP arm, 47.7% in the GP arm and 42.9% in the DP arm (P > 0.05). The response rate of patients with radiochemotherapy was 69.9%, and 40.8% for those with chemotherapy alone (P < 0.05). In group NP, TP, GP and DP, median TTP was 5.7, 5.3, 5.9 and 5.5 months (P > 0.05) respectively, MST was 10.4, 10.6, 11.5 and 10.4 months (P > 0.05) respectively, 1-year survival rate was 41.9%, 41.1%, 43.1% and 42.9% (P > 0.05) respectively, and 2-year survival rate was 21.3%, 19.4%, 23.1% and 23.2% (P > 0.05) respectively.

CONCLUSIONSThe third-generation cisplatin-based chemotherapy regimens may be the standard treatment for advanced NSCLC, and their combination with radiotherapy may improve the therapeutic efficacy and prolong the survival of patients.

BACKGROUNDTo observe and compare the efficacy and safety of IEP and EP regimens for small cell lung cancer (SCLC).

METHODSSixty-four patients with SCLC pathologically proved were randomly divided into IEP group ( n =32) and EP group ( n =32).

RESULTSAll the 64 patients were evaluable for response and toxicity. In IEP group, the total responsive rate, responsive rates of limited-stage patients and extensive-stage patients were 84.4%(27/32), 100.0%(15/15) and 70.6%(12/17) respectively; while in EP group, those were 75.0%(24/32), 85.7%(12/14) and 66.7% (12/18) respectively. The median duration of remission was 6 months and 1-year survival rate was 62.5% in IEP group, and 5 months and 56.2% in EP group. There was no significant difference in response rate, median duration of remission and 1-year survival between the two groups ( P > 0.05). The main toxicity was myelosuppression. Incidences of leukopenia at grade III-IV, nausea, vomiting and alopecia were significantly higher in the IEP arm than those in the EP arm ( P < 0.01 ).

CONCLUSIONSHigh response rates and tolerable toxicities are attainable for small cell lung cancer treated with IEP and EP. IEP regimen shows a similar response rate compared with EP regimen. They might be considered as relevant regimens in initial patients with small cell lung cancer.

OBJECTIVETo explore the practicability of microskin grafting by spraying in burn management.

METHODSRazor thin autologous skin from pigs or burn patients was harvested and cut to pieces of 0.2 - 0.5 mm in size and suspended in normal saline. The suspension was put into a bottle with outlet and pumping device. The microskin suspended in the saline was sprayed to the burn wound and/or onto the alloskin sheets. The microskin distribution was detected by digital image analysis technique. In animal experiments, the burn wound development and pathomorphological changes after operation were observed. In burn patients who would receive microskin grafting, spraying method was used with the traditional flotation method as control. The treatment results and the operational procedures were compared between these two kinds of operation styles.

RESULTSThe microskin dispersion degree with spraying was much smaller than that with flotating method. In animal experiment with spraying method, the wound healing time was 23.2, 24.5 and 38.3 days in 3 groups, respectively, with the area ratio of donor to wound of 110, 120 and 150. In clinical study, The average operating time was 133.3 min with spraying method and 165.6 min with flotation method respectively (P < 0.05). The area ratio of donor to wound was 118.8 with spraying and 17.6 with flotation methods, respectively (P < 0.01). The one time wound coverage rate was 92.6% with spraying and 79.7% with flotation methods (P < 0.01). The wound healing time was 29.7 days with spraying and 37.3 days with flotating methods, respectively (P < 0.05).

CONCLUSIONSpraying method of microskin grafting might be a good method in major burn treatment. The advantages with this method included well-distributed microskin, simpler handling, saving of donor skin, shortening of operating time and less time needed for the wound healing. It might be recommended for other wound covering materials.

Animals ; Burns ; surgery ; therapy ; Culture Techniques ; Dermatologic Surgical Procedures ; Female ; Humans ; Male ; Rabbits ; Skin ; injuries ; Skin Transplantation ; methods ; Transplantation, Heterologous ; Wound Healing