Objective To evaluate the performance of enzyme linked immunosorbent assay (ELISA) kit in detection of Hepatitis B virus(HBV) markers by using improved and optimized method ,so as to provide a practical and feasible method and reagents for clinical laboratory .Methods ELISA test was used for the detection of HBV markers .The gradient dilution method was used to e‐valuate the lower limit .The verifiation of cut off value was carried out based on clinical and laboratory standards institute (CLSI) EP12‐A2 document .Samples with cut off values were collected to evaluate the precision ,including repeatability and intermediate precision .The coincidence rates were counted through comparing the results of ELISA with those of external quality assessment and those detected by using Abbott i4000SR chemiluminescence instrument .Results The lower detection limit of HBsAg ,HBsAb , HBeAg ,HBeAb and HBcAb were 0 .2 IU/mL ,20 mIU/mL ,1 NCU/mL ,0 .75 NCU/mL and 0 .05 NCU/mL respectively .The cut‐off value(C50 )± 20% concentration included the concentration range between C5 and C95 .The within‐run coefficient of variation (CV)≤15% ,in sandwich method the between‐run CV≤25% ,in competition method the between‐run CV≤35% .The positive and negative coincidence rates in accuracy and comparing with i 4000SR all were more than 95% and all κ>0 .75 .Conclusion ELISA tests for HBV markers in our laboratory could meet the requirements of the detection performance and clinical needs .

The expressing cassette, LoxP-CMV-gpt-IRES-LoxP( about 2.9kb), was amplified by PCR from a plasmid, pIRES-gpt, by use of the primers , which contained the loxP sites in 5' terminals, respectively. The loxP sites were designed into primers by the software of Primer primer 5.0. Then the cassette was cloned into the site of BalI in pBUS10 to obtain pUS-gptIRES(L). The sequencing analysis for pUS-gptIRES(L) indicated that two loxP sites with the same direction were correctly inserted into pUS-gptIRES(L).The gpt gene in pUS-gptIRES(L) was replaced by a fragment including the full length GFP gene as well as SV40 poly A sequence to get pUS-GFPIRES(L). pUS-GFPIRES(L) was transiently transfected into CHO cell lines, and then the green fluorescence could be seen, the results showed that GFP gene could be expressed correctly. Moreover, pUS-GFPIRES(L) was transfected into the CEF infected MDV CVI988 strain and recombinant virus was selected by the green fluorescence. The growth curve of virus showed the characteristic of recombinant virus was the same as that of CVI988 in vitro. These results give the basis for further studying the characteristic of MDV in vivo and the application of the Cre/LoxP system to MDV genome.

OBJECTIVETo observe the effect of Ginkgo Leaves Tablet (GLT) on memory quotient (MQ) of mild cognitive impairment (MCI) patients.

METHODSOne hundred and thirteen patients were randomly assigned to the control group (55 cases) and the treatment group (58 cases). Patients in the control group received dietetic therapy and physical exercises, while those in the treatment group additionally took GLT, 19.2 mg each time, three times daily. The treatment course was 12 months for all. The MQ of all the patients was assessed by WMS-RC before treatment,at 6-month of treatment, and 12-month of treatment.

RESULTSCompared with the control group, the improvement of MQ increased in the treatment group 0.5 and 1 year after treatment (P < 0.05). The clinical efficiency of MQ obviously increased in the treatment group (48.28% and 50.00%), showing statistical difference when compared with the control group (30.91% and 27.27%, P < 0.05, P < 0.01). There was statistical difference in added scores of recognition, regeneration, understanding, and recitation test at 6-month of treatment and 12-month of treatment between the treatment group and the control group (P < 0.05, P < 0.01).

CONCLUSIONGLT was effective in improving MQ of MCI patients, especially in improving recognition, regeneration, understanding, and recitation test.

Aged ; Aged, 80 and over ; Cognitive Dysfunction ; drug therapy ; psychology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ginkgo biloba ; chemistry ; Humans ; Male ; Memory ; Middle Aged ; Phytotherapy

A novel white-rot fungus AH28-2,which was isolated from 224 fungi samples,ability to produce effectively laccase by induction.Several factors influencing laccase production were investigated.The optimum conditions were as follows:the 300mL Erlenmeyer flask containing 150mL of liquid medium was inoculated with 7.5mL of mycelial fragments and the medium was supplied with lignin at a concentration of 0.1%(initial pH8.5).The cultures was incubated at 28℃ on rotary shaker(150r/min) for 4～5 days.The maximum enzyme activity was 20184IU/L.

OBJECTIVETo construct wild-type and mutant pEGFP SPAST vectors and to explore the molecular mechanism of hereditary spastic paraplegia.

METHODSMutant SPAST vector was constructed using overlap PCR method following construction of wild-type SPAST vector. Wild-type and mutant constructs were transfected to COS7 cells and subcellular localization of spastin was observed. Co-localizations of spastin and microtubule, spastin and mitochondria were viewed by immunofluorescence staining.

RESULTSWild-type spastin is localized in plasma, and mutant spastin did not change its cellular localization. Wild-type and mutant spastins did not co-localize with microtubules and mitochondria by immunofluorescence analysis.

CONCLUSIONWild-type and mutant SPAST constructs were successfully generated. Mutant spastin did not change its localization in cells. Spastin does not co-localize with microtubules and mitochondria. This study may facilitate further studies on molecular mechanism of hereditary spastic paraplegia.

Adenosine Triphosphatases ; genetics ; metabolism ; Animals ; Base Sequence ; Cell Line ; Genetic Vectors ; genetics ; Humans ; Mitochondria ; genetics ; metabolism ; Mutation ; Spastic Paraplegia, Hereditary ; genetics ; metabolism ; Spastin

BACKGROUNDTo evaluate the safety of intracytoplasmic sperm injection (ICSI) with epididymal or testicular sperm, this study compared children born after ICSI treatment with epididymal or testicular sperm with children conceived after ICSI with ejaculated sperm.

METHODSThis retrospective study included 317 children born after ICSI with percutaneous epididymal sperm aspiration (PESA), 103 children born after ICSI with testicular sperm aspiration (TESA), and a control group of 1008 children born after ICSI with ejaculated sperm. All of the patients received their assisted reproductive treatment in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2004 to December 2011. Data, such as the rate of stillbirths, perinatal mortality, gestational age, birth weight, and the rate of congenital malformations of the three groups, were compared.

RESULTSPESA and TESA children were not different from ICSI children in the rate of stillbirths, perinatal mortality, infant mortality rate, gestational age, the rate of prematurity, and the rate of malformations (P > 0.05). A slight increase in birth defects was reported in the TESA group compared with those in the control group, but there was no significant difference between the groups (P > 0.05).

CONCLUSIONICSI with epididymal or testicular sperm does not lead to more stillbirths or congenital malformations compared with ICSI using ejaculated sperm.

Adult ; Congenital Abnormalities ; epidemiology ; Epididymis ; Female ; Fetal Death ; epidemiology ; Follow-Up Studies ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Retrospective Studies ; Sperm Injections, Intracytoplasmic

AIM: To report the long- term clinical outcomes of accelerated trans-epithelial corneal cross-linking ( CXL ) protocols using KXL System ( Avedro, USA ) in the treatment of progressive keratoconus. · METHODS: Totally 52 patients ( 102 eyes ) with progressive keratoconus between December 2014 and February 2017 [ maximum keratometry values ( Kmax) ≤60.0D, minimum corneal thickness(Thk) ≥400m] were treated with an accelerate trans-epithelial CXL protocol (UV-A irradiation intensity 45mW/cm2 with a total fluence of 7. 2J/cm2 ) using KXL system ( Avedro, USA ) in Southwest Hospital. The average follow-up time was 11. 65mo (range: 9-26mo). Uncorrected distance visual acuity ( UDVA) , corrected distance visual acuity ( CDVA) , intra- ocular pressure ( IOP ) , slit-lamp microscope examination, Kmax and average keratometry values ( AveK ) , corneal stromal demarcation line depth and endothelial cell density ( ECD) were evaluated. ·RESULTS:The 52 patients (102 eyes) were included in this research, male 36 (70 eyes) and female 16 (32 eyes), average age was 19. 5±4. 6 years. Preoperative CDVA was 0. 84±0. 89 (LogMAR), postoperative CDVA was 0. 69±0. 72 ( P = 0. 398 ). Preoperative UDVA was 1. 02 ± 0. 62 (LogMAR), postoperative UDVA was 0. 85 ± 0. 59 ( P =0. 154 ). Preoperative IOP was 12. 95 ± 4. 40mmHg, postoperative IOP was 11.92±3. 66mmHg (P=0. 272). No statistical difference (P=0. 552) has been found between preoperative and postoperative ECD. Nevertheless, on the Sirius anterior system ( Sirius, CSO, Itlay) , significant statistical difference (P=0. 017) was confirmed between preoperative Kmax ( 50. 83 ± 3. 48D ) and postoperative Kmax (52. 05±3. 63D). Meanwhile, the postoperative Avek (47.74±2. 51D) was significantly lower (P=0. 041) than the preoperative Avek ( 48. 73 ± 4. 33D ). The average corneal stromal demarcation line depth ( 192 ± 23. 6μm ) was detected by the anterior segment OCT. No statistical difference ( P = 0. 816 ) has been found between preoperative and postoperative Thk. No severe complication was observed in all cases. ·CONCLUSION: Accelerated trans-epithelial CXL was effective in decreasing keratometry values for progressive keratoconus in this research, and the outcomes remained stable during the follow-up time. No endothelium damage or other severe complications were observed in this clinical research. The accelerated trans-epithelial CXL is as effective as the standard CXL.

OBJECTIVETo analyze the genetic polymorphism of 15 single nucleotide polymorphism (SNP) loci on the nonrecombining portion of the Y chromosome in 6 populations in China.

METHODSAllelic specific polymerase chain reaction and 2% agarose gel electrophoresis and 6% PAGE were used to analyze the genetic polymorphism of 343 unrelated males, representing 6 populations in China, including Fujian Hans, Sichuan Hans, Mongolian, Hezhen, Sibo and Hui from the South, Northeast and Northwest.

RESULTSThirty haplogroups were observed, and 3 of them (H15, H16, H18) were seen in all of the six populations. Although the heterozygosity levels of the Hezhen, Mongolian, Sibo populations are similar and those of the other 3 populations (Fujian Hans, Sichuan Hans, Hui) are similar, the pairwise differences among haplogroups are significant. Analysis of molecular variance (AMOVA) and principal component (PC) analysis of the haplogroup distributions suggested highly different allele diversity between group I including Hezhen, Mongolian, Sibo and group II including Hui, Fujian Hans, Sichuan Hans.

CONCLUSIONThe above analyses show more significant variance components in Northeast/South populations and clearly reveal the geographic genetic relationship among the six populations in the Northeast/Northwest/South. These results confirm the complexity of the genetic structure of Chinese populations and make a significant contribution for constructing the contemporary human gene pool and tracing genetic dispersal trail from Chinese populations.

Alleles ; China ; ethnology ; Chromosomes, Human, Y ; Genetic Variation ; Genetics, Population ; Humans ; Polymorphism, Single Nucleotide

OBJECTIVETo determine the effects of modified pull-through operation (Badenoch operation) on the treatment of posterior urethral stricture.

METHODSFrom September 2001 to December 2010 traditional pull-through operation was Modified for two times in our center. A total of 129 patients with posttraumatic posterior urethral stricture resulting from pelvic fracture injury underwent the modified urethral pull-through operation. Stricture length was 1.5 to 5.3 cm (mean 2.9 cm). Of the patients 43 had undergone at least 1 previous failed management for stricture. In phase 1 (from September 2001 to January 2008), the improving items include: (1) The distal urethral end was stitched and tied to the catheter. (2) As catheter was inserted into bladder and 20 ml water was injected into catheter balloon, the distal urethral end was fixed in the proximal urethra and an overlaying of 1.5 cm was formed between the two ends. (3) Three weeks later, it was tried to insert the catheter to bladder. After the urethral stump necrosis and the catheter separating from the urethra, the catheter was removed. In phase 2 (from February 2008 to December 2010), based on the above, irrigating catheter was used. After the surgery, urethra was irrigated with 0.02% furacillin solution through the catheter 3 times a day. All patients were followed up for at least 6 months. If patients had no conscious dysuria and maximum urinary flow rate (Qmax) > 15 ml/s, the treatment was considered successful. All complications were recorded.

RESULTSIn phase 1, the 96 patients (101 times) underwent the procedure. The treatment was successful in 88 patients (success rate 92%). Within 1 to 13 days after removal of the catheter, urethral stricture was recurred in 8 patients. They had to undergo cystostomy once more for 3 to 11 months before reoperation (the 3 patients' reoperation was in phase 2). The 8 cases were treated successfully. In phase 2, 33 patients (total 36 times) underwent the procedure. One patient was failed (success rate 97%). The actual follow-up time is 7 to 93 months (An average of 37.6 months). Qmax is (22 ± 5) ml/s. No complications such as urinary incontinence, erectile pain, urinary shortening happened.

CONCLUSIONSThe modified urethral pull-through operation is effective for the surgical treatment of posttraumatic posterior urethral stricture. It has a high success rate with durable long-term results. Complications are few. The procedure is simple, less demanding and especially suitable in patients who had previously undergone failed surgical treatments.

Adult ; Aged ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Urethra ; surgery ; Urethral Stricture ; surgery ; Young Adult

AIMTo study the permeability of nerve growth factor (NGF) liposomes (NGF-L, NGF-SSL, NGF-SSL-T) on the blood-brain barrier (BBB) model and the distribution in vivo, and analyze the correlation between the results in vitro and in vivo.

METHODSThe BBB model in vitro was established by using mouse brain microvascullar endothelial cell, and the model was applied to study the permeability of NGF liposomes. The distribution of NGF of each group was studied by 125I labeled and SDS-PAGE method.

RESULTSThe highest encapsulation proportion was 34%, and the mean size of NGF liposomes was below 100 nm. The permeability of NGF liposomes on in vitro BBB model showed that the liposome could promote NGF to transport across the BBB, the permeability of NGF-SSL-T was the highest. The distribution in the brain showed in an order of NGF concentration NGF-SSL-T > NGF-SSL + RMP-7 > NGF-SSL > NGF-L. There was a close relationship between P(e) (permeability coefficient on in vitro BBB model) and BUI (brain uptake constant in vivo).

CONCLUSIONLiposomes can promote NGF to transport across the BBB, and the transporting ability BBB of NGF-SSL-T which RMP-7 incorporated into the surface of NGF liposomes is the best.

Animals ; Biological Transport ; drug effects ; Blood-Brain Barrier ; Bradykinin ; analogs & derivatives ; pharmacology ; Brain ; metabolism ; Cell Membrane Permeability ; Drug Delivery Systems ; Endothelial Cells ; cytology ; Liposomes ; Male ; Mice ; Nerve Growth Factor ; administration & dosage ; pharmacokinetics ; Particle Size ; Rats ; Tissue Distribution