BACKGROUND:Bulous keratopathy has an increasing annual incidence, but its treatment is restricted by few sources of materials for corneal transplantation and high cost of operation. Additionaly, some patients who present with serious symptoms have little chance of recovery and low success in corneal transplantation. Amniotic membrane from the corneal stroma has a rich source with low cost, which can effectively relieve the symptoms and improve the quality of life in patients.
OBJECTIVE:To observe the therapeutic efficacy of amniotic membrane implantation into the corneal stroma in the treatment of bulous keratopathy.
METHODS: Forty healthy adult New Zealand rabbits (half male and female) were randomly divided into four groups (A, B, C, D groups), with 10 rats in each group. Rabbit models of bulous keratopathy were made in the groups A, B, C. At 2 weeks after modeling, amniotic membrane implantation into the corneal stroma and corneal surface was performed in groups A and B, respectively, and in group C, corneal lamelar dissection was done but with no amniotic membrane transplantation. In group D, there was no surgical treatment (blank control). A slit lamp microscope with constant crack width and angle of light projection was used to observe the central corneal thickness, and corneal opacification degree, corneal epithelial bula of rabbits were observed at different time in each group. Under microscope, the rabbit corneal endothelial cels and healing were observed at different time.
RESULTS AND CONCLUSION: At 1 day and 2 weeks after transplantation, the central corneal thickness of rabbits had significant differences in the four groups (P< 0.05). At 4, 8, 12 weeks after transplantation, the central corneal thickness of rabbits showed no difference between groups A and B as wel as between groups C and D (both P > 0.05), but there was a significant difference between groups A, B and group C (P < 0.05). At 4 and 8 weeks after transplantation, the degree of corneal opacity was significantly better in group A than the other three groups (P < 0.05). There were obvious scars forming at the incision of rabbits in the group C. Compared with the other three groups, the bula was improved better in the group A (P < 0.05). At 2 weeks after transplantation, bulous keratopathy relapsed in the group B, and symptoms of edema with bula were stil seen in groups C and D at 12 weeks after transplantation. These findings indicate that amniotic membrane implantation into the corneal stroma can effectively repair rabbit corneal endothelial cels and aleviate the symptoms of edema, but its specific mechanism need to be further studies.

Objective To explore immunological mark and the outcome of disease after pharmacological treatment in human NCC with praziquantel. Methods 35 patients were treated with praziquantel for 6 months. Levels of serum IL-2,IFN-γ, IL-12, IL-5, IL-10 and TNF-α were measured before and after treatment. Results Th1 type cytokines IL-2, IFN-γand IL-12 were up regulated after treatment( P ＜ 0. 01 ) . Levels of TNF-α and Th2 type cytokines IL-5 and IL-10 were remarkable decreased after treatment( P ＜ 0. 01 ). The levels of IL-2, IFN-γ and IL-12 from the patients with good response to the treatment is higher than those with no response to the treatment. Conclusion Th1 type cytokines were up regulated while Th2 type cytokines were down regulated in the NCC patients treated with praziquantel. The protective immunity may be related to the Thl cell activation.

Objective To establish real-time PCR method for the detection of JAK2 V617F mutation and evaluate its clinical significance in patients with myeloprollferative disorders and leukemia.Methods 71 chronic myelocytic leukemia(CML) patients, 22 essential thrombocythemia (ET) patients, 11 primary myelofibrosis (PMF) patients, 9 polycythemia vera (PV) patients and 7 cosinophilia patients were enrolled in this study. JAK2 V617F mutation was determined by real-time PCR and amplification refractory mutation system (ARMS), followed by sequencing. Human erythroleukemia cell (HEL cell)DNA was used as homozygous control of JAK2 V617F mutation. The detection limit for either real-time PCR or ARMS was evaluated. Results Real-time PCR assay showed that there was a melting temperature(Tin) peak at (75.0±0.2)℃ for wild type samples and a Tm peak at (76.6±0.2)℃ for mutation type samples. JAK2 V617F mutation was detected in 8(88.9%) patients with PV, 12(54.5%) patients with ET and 7(63.6%) patients with PMF respectively. But there was only one positive case in 71 CML patient (1.4%). The results showed complete concordance with ARMS results and confirmed by sequencing. The mutation could be detected in 102 HEL cells per 106 white blood cells by real-time PCR, whereas the mutation can be assessed in 104 HEL cells per 106 white blood cells by ARMS. Thus, the sensitivity of real-time PCR was 100-fold higher than ARMS. Conclusions The real-time PCR method is successfully established for detection of JAK2 V617F mutation. This method is more sensitive, convenient than ARMS, and suitable for clinical application. There is high frequency of JAK2 V617F mutation in myeloproliferative disorders and it could be used as the diagnostic marker for myeloproliferative disorders.

OBJECTIVE: To study the inhibitory effects of traditional chinese medicine Compound Liuyuexue (CLYX) on hepatitis B surface antigen(DHBsAg) and hepatitis B e-antigen(DHBeAg).METHODS: One-day old guangxi brown spotted ducks infected with DHBV were used as the hepatitis B virus infected animal model. Positive ducks were detected by PCR at 13 days after the infection of DHBV, and were randomly divided into five groups: the high dose group, middle dose group and low dose group of Compound Liuyuexue(CLYX), model group, and positive control group, with 10 ducks in each group. CLYX was given ig.for 14 days. Serum sampling was scheduled at 0, 7, 14 days respectively and 3 days after drug withdrawal, and the contents of DHBsAg and DHBeAg in serum were measured by ELISA. RESULTS: The serum DHBsAg and DHBeAg contents in high dose and middle dose groups of CLYX were decreased significantly(P

AIM:To study the inhibitory effect of Compound Liuyuexue(Tarphochlamys affinis(Giff) Bremekhu,Herba Hedyotis,Raidx Gardeniae,etc.)(CLYX) on HBV in vitro.METHODS:The serum containing CLYX was added to cultured HepG2.2.15 cells,and HepG2.2.15 cells were cultured in medium containing the serum with CLYX for 72 h and 144 h.The culture media were collected for determining the levels of HBsAg and HBeAg by ELISA.RESULTS:The serum containing CLYX could markedly inhibit HBsAg and HBeAg expressions in the HepG2.2.15 cells(P

Aim To observe the effects of LYS polysaccharides on expressions of APP, PS1, PS2 and ApoE in SAMP8 mouse brain. Methods 50 6-month-old SAMP8 mice were used and divided randomly into 5 groups: SAMP8 untreated control group, huperzine A control group, low-, mid-and high-dose groups of polysaccharides, with 10 mice in each group. 10 6-month-old SAMR1 mice were used as normal control. After each group was treated by corresponding drug for 40 days continuously, the mRNA contents of APP, PS1, PS2 and ApoE in brain tissue were assayed by reverse transcription polymerase chain reaction.Results The low-, mid-and high-dose groups of polysaccharides could reduce the mRNA contents of APP, PS1 and PS2, which showed a dose-effect relationship in some degrees. But it could not affect the content of ApoE. Conclusion LYS polysaccharides could inhibit the expressions of APP, PS1, and PS2 in SAMP8 mouse brain.

AIM:To study the isolation and composition of Cliffbean(Millettia pulchra kurz)polysaccharide.METHODS:Cliffbean was extracted by boiling water.The polysaccharide in the filtrate was precipitated fractionally by alcohol.The protein in the product was removed by Sevag method,and was further purified by DEAE ion-exchange cellulose(DEAE-52).The component of the saccharide was determined by GC and TLC.RESULTS:The polysaccharide obtained in this way didn't contain protein and nucleic acid.It showed a single polysaccharide when identified by means of Sephadex G-75 column or High Performance Gel Permeation Chromatography(HPGPC).It was composed of glucose and arabinose.CONCLUSION:Cliffbean polysaccharide was first isolated from its root and was one of the important components.

Objective To investigate a new single nucleotid polymorphism (SNP) intron5(+4668C/T) in SLC22A12 in primary gout patients and the association between clinical characteristics and genotypes. Methods One hundred and one primary gout patients and 186 healthy subjects were recruited into this study. Blood pressure, body mass index (BMI) was recorded. Serum uric acid, glucose, lipid and creatinine were detected. DNA was extracted from peripheral blood to amplify the fragment located in intron 5. The genotypes of SLC22A12 can be detected with high-resolution melting (HRM) assay, followed by sequencing analysis. Chi-square test was used for statistical analysis. Results ① A new SNP in intron 5 of SLC22A12 was identi-fied successfully by HRM, which was defined as intron 5 (+4668C/T). CC, CT and TT genotypes were unam-biguously distinguished with HRM technology, which was fully concordant with sequencing. ②The genotypes of CC, CT and TT in male and female groups were 28.1%, 33.7%, 38.2% and 20.0%, 47.1%, 32.9%, respectively.③ However, no significant differences of genotype distribution were found concerning BMI, blood pressure, creatinine, total cholesterol and triglyceride in both male group and female group. But the serum uric acid levels in the CC genotype were significantly higher than those with the CT+TT genotypes. ④ The genotype frequencies of CC and CT+TT in high uric acid group were remarkably different from those in low uric acid group (21.2%, 78.8%,; 35.0%, 65.0%; P<0.05). Conclusion A new SNP has been successfully discovered with HRM technology with simplicity, rapidity and accuracy. T allele of intron 5 (+4668C/T) may be a genetic protective factor for hyperuricemia among Chinese population.

Objective To investigate the relationship between HLA-B * 5801 allele and severe cutaneous adverse reactions caused by allopurinol or other drugs.The clinical value of HLA-B * 5801 as the marker of allopurinol-SCAR was evaluated.Methods Forty-three patients with allopurinol-SCAR,133 patients without SCAR after taking allopurinol for 3 months were included.Ninety-six patients with SCAR caused by other drugs and 148 healthy individuals were enrolled into the present study.HLA-B * 5801 allele was detected by PCR-SSP method.Data were analyzed by chi-square test.Results HLA-B * 5801 was present in 40 of 43 (93.0％) patients with allopurinol-SCAR,which was significantly higher than 19 of 148 (12.8％) in healthy subjects (x2=100.353,P＜0.01,OR=90.5,95％CI 25.5-321.8).But there were no significant differences between allopurinol-tolerant patients and healthy controls(10 of 133,7.5,x2=2.141,P＞0.05,OR=0.6,95％CI 0.2-1.2).And there were only 14 of 96 (7.5％) patients with SCAR caused by other drugs had HLA-B * 5801 (x2=0.152,P＞0.05,OR=1.2,95％CI 0.6-2.4).Conclusion The study indicates that people with HLA-B * 5801 have a high risk of allopurinol-SCAR.HLA-B * 5801 is a specific and predictive marker for guiding the selection of uric acid lowing drug allopurinol.

Long non-coding RNA (lncRNA) is defined as non-protein coding transcript longer than 200 nucleotides. In the form of RNA, it affects gene expression at the epigenetic, transcriptional and post-transcriptional levels, and is widely involved in the body's pathophysiological processes. This review summarizes the research progress of lncRNA in the field of parasitology in order to find new targets for the prevention and treatment of parasitic diseases.