Enterovirus A, Human ; Hand, Foot and Mouth Disease ; virology ; Humans

Objective To observe the expression of osteoprotegerin (OPG), receptor activator of nuclear factor κβ ligand(RANKL) and receptor activator of nuclear factor κβ (RANK) in bone tissue of rats with chronic fluorosis and to explore the relation between OPG/RANKL/RANK system and bone damage in chronic fluoride poisoning rat and the antagonism effects of Danlan Xianpeng capsule. Methods SD rats were randomly divided into six groups according to body weight (equal male and female in each group): fluorosis group, high dose drug group, medium dose drug group, low dose drug group, control group, borax group(positive control), 12 rats in each group. The control group drank tap water and the remaining 5 experimental groups consumed 50 mg/L fluoride water, and high, medium and low doses drug group took Danlan Xianpeng capsule at doses of 0.8,0.4,0.2 g/kg,borax group took borax at dose of 0.8 g/kg. OPG, RANKL, RANK protein in rat tibial metaphysis was detected by immunohistochemistry at the 6 month. Results Compared with the control group(173.79 ± 5.23, 174.17 ± 5.01,155.63 ± 7.11), the expressions of OPG, RANKL were increased and the expression of RANK was decreased in fluorosis group(156.83 ± 5.80, 157.74 ± 6.70, 173.92 ± 4.37), the difference was statistically significant (all P ＜ 0.05). Compared with the fluorosis group, the expression of OPG and RANKL were decreased and the expression of RANK was increased in high-dose drug group, middle-dose drug group(169.67±5.07, 168.08 ± 5.05,162.12 ± 4.24, 170.78 ± 5.01, 168.41 ± 7.19, 166.69 ± 5.78, all P ＜ 0.05). Compared with the borax group (167.27 ± 4.08, 167.85 ± 5.01, 166.14 ± 3.95), the expression of OPG and RANKL was increased in the low-dose drug group (163.40 ± 4.11, 159.49 ± 5.78), the expression of RANK was increased in the high-dose drug group (162.12 ± 4.24) and decreased in the low-dose drug group(171.54 ± 8.06), the difference was statistically significant (all P ＜ 0.05). Conclusions Chronic fluoride poisoning can cause increased bone turnover and enhance the activity of osteoelastic absorption by increasing RANKL. Danlan Xianpeng capsule can affect bone remodeling through the OPG/RANKL/RANK system, and antagonises bone damage caused by fluoride.

At the beginning of the 1980s, a concept of viable but non-culturable(VBNC) was suggested. VBNC is a survival strategy adopted by microorganisms when they are exposed to environmental stress. This article try to make a summary of research of the conditions of VBNC formation, recovery of culturability and methods of VBNC cells detection. In addition, introduces the first growth factor of microorganisms-Rpf.

Hematologic Diseases ; Hemolysis ; Humans ; Infant, Newborn

This project is targeted on exploring some improving approaches to isolate and culture the microorganisms which are difficult to be isolated and cultured through the conventional ways. The results showed that betaine, sodium pyruvate, SOD and catalase are helpful for increasing the total number and variety of isolated strains. A kind of combined method was also used to isolate the micro-colony which can not be seen by naked eyes on the plates. Totally 52 Actinomycetes and 103 bacteria and 17 fungi were obtained from 4 soil samples using the above methods. 4. 325% microorganisms were obtained as positive strains to inhibit the growth of some kinds of test bacteria, which is higher than the percent using generally isolated ones. These microbial natural products may remain an important resource for the drug discovery.

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

To decrease the oxygen content in the cell is a key method to improve hydrogen production in Chlamydomonas reinhardtii.A new approach was developed by transforming the leghemoglobin gene lba,which has high affinity to oxygen,into the chloroplast of C.reinhardtii to get a low dissolved oxygen in the cell and result into improvement of H2 ase activity and H2 yield. The results showed that lba was successfully transformed into the chloroplast of C.reinahrdtii strain 849 and did not affect its growth significantly. The work paved the road for further regulation of lba expression in the chloroplast to improve of hydrogen production of C.reinahrdtii.

Objective To amplify all immunoglobulin V genes of human atherosclerosis by PCR and to sequence the products of PCR cloned into T-vector. Methods Peripheral blood samples of 50 patients with atherosclerosis were collected,from which lymphocytes were segregated by density gradient centrifugation and total RNA was extracted by TRIzol reagent.V genes of VL(including VK and V?) and VH were amplified by RT-PCR,and the products were digested by restriction enzyme and then cloned into T-vector.Two clones were picked randomly to sequence.(Results)Total RNA were extracted purely with integrity,and all V genes of VL and VH were amplified by PCR successfully.The sequences were highly homologous to human immunoglobulin genes. Conclusion All immunoglobulin V genes of human artherosclerosis were amplified successfully,which lays a foundation for the construction and (selection) of phage library.

AIM: To study the causes of pyogenic granuloma after hydroxyapatite(HA) orbital implants.METHODS: HA orbital implants (250 cases) in our hospital (68 pegged implants) were reviewed.All patients were followed up from 18 months to 10 years. Implants were removed after medical therapy which was proved to be ineffective.RESULTS: Ten of 250 cases of HA orbital implants developed pyogenic granuloma. Pyogenic granuloma occurred in 1 unpegged implants patient and 9 patients after pegging and drilling of HA implantation over 4～7 years. The pyogenic granulomas were not controlled by medical therapy effectively. Implants were removed in 9 cases except 1 case denied removing and continued medical therapy.CONCLUSION: Pyogenic granuloma was serious complication that occurred after HA orbital implants. Partial vascularization, implant exposure, xenogenic sclera implant, pegging and drilling of HA implantation are risk factors that affect the development of pyogenic granuloma.Pyogenic granuloma hasn't relation with implanted peg material. Pyogenic granuloma denotes the potential implant infection, and all implants should be removed finally.

To evaluate the value of silicone intubation and ring-fixed method in canalicular laceration.
●METHODS: A retrospective study of 36 cases with laceration of lacrimal canaliculus. For all the patients, microscope was used to find the broken ends of the lacrimal canaliculus, and then straight insertion was performed to the distant and near broken ends through the upper and lower lacrimal points, then passed the dacryocyst to insert nasolacrimal canal and ended up in inferior nasal meatus. Thus, clinical data about forming annular support to treat laceration of lacrimal canaliculus was presented here.
● RESULTS: All 36 patients with traumatic canalicular laceration were anastomosed successfully and all patients healed without infection. The surgery went well, intubation was smoothly, all patients were followed - up for another 6mo after the stent was removed. The accidental fall off of the stent was not observed during the follow - up. During the follow - up 10 - 18mo after extubation, when the stent was removed, 32 cases (88. 9%) recovered to normal lacrimal drainage function;4 cases ( 11. 1%) remained epiphora with obstructed lacrimal passage. All cases were no lower eyelid ectropion.
●CONCLUSlON: For the patients with inferior canalicular laceration, the silicone intubation and ring-fixed method is effective, low irritation and less complication.