Objective To provide a technique of preparation recombinant hALR by gene engineering in larger scale.Methods Construct hALR(human augmenter of liver regeneration) protein expression vector pGEX-3X-hALR,induce the expreesion of hALR,and pruified the protein by affinity chromatography.Results Succeed in constructing the vector pGEX-3X-hALR and obtained the recombinant hALR protein.Conclusion The mothed could be used for preparation recombinant hALR by gene engineering in larger scale.

BACKGROUND:It has been reported that treatment with granulocyte-colony stimulating factor (G-CSF) increases the abundance of circulating CD34+ cells in rats. Data from the study, more important, suggested that mobilized by G-CSF may enhance rapid reendothelization and reduce neointimal formation after vascular injury.OBJECTIVE: To evaluate whether BM-derived CD34+ cells could enhance rapid reendothelization and reduce neointimal formation after balloon-injured carotid artery in an intact rat model.DESIGN: Randomized control animal study.SETTING: Nanjing First Hospital Affiliated to Nanjing Medical University.MATERIALS: The experiment was carried out in Nanjing First Hospital from December 2005 to April 2006. A total of 40 male Sprague-Dawley rats, weighing 200-250 g and of SPF grade, were purchased from National Rodent Laboratory Animal Resources, Shanghai Branch. The recombinant human G-CSF was purchased from Qilu Pharmaceutical. The 2F Fogarty arterial embolectomy catheters were purchased from Edwards Lifesciences. Anti-human CD34 and anti-human CD45 were purchased from Multi Sciences.METHODS: SD rats were divided randomly into treated group (n =20) and control group (n =20). Subcutaneous injection of recombinant human G-CSF (100 μg/kg/day) once daily for 8 days for treated group. Control group as treated with subcutaneous injection of saline. Five days after initiation of G-CSF treatment or saline, the rats were anesthetized by intraperitoneal injection with ketamine. The left common carotid artery was exposed through a midline incision of the ventral side of the neck. A 2F Fogarty arterial embolectomy catheter was inserted through the external carotid artery,inflated with 200 μL air, and passed 3 times along the length of the segment, which was defined proximally by the carotid bifurcation and distally by the edge of the omohyoid muscle. After removal of the catheter, the proximal ligature of the external carotid artery was tied off. ① An average of 1 mL venous blood per rat was collected for enumeration of the white blood wells (WBCs) and CD34+ cells before and 5 days after initiating G-CSF or saline treatment. ② Ten rats in each group were killed with overdose ketamine at 14 and 28 days after balloon injury and left common carotid arteries were harvested. The luminal surface of carotid arteries (n =5, each group) was exposed to calculate the reendothelialized area, which was manually traced with software (Image ProPlus). Reendothelialized area = non-stained with Evans blue area/the total area of balloon-injuried. The cross sections of carotid arteries (n =5, each group) were stained with hematoxylin and eosin (HE) and calculated intima-to-media area ratio (I/M) with software (Image ProPlus) to assess the extent of neointimal thickening. ③ To evaluate the extent of reendothelialization of arteries injury, sections were stained with CD31 and vWF by immunohistochemistry analysis.MAIN OUTCOME MEASURES: ① The number of WBCs and CD34+ cells; ② the extent of reendothelialization of arteries injury; ③ the extent of neointimal hyperplasia (I/M); ④ CD31 + and vWF+ endothelial cells.RESULTS: A total of 40 rats were involved in the final analysis. ① The number of WBCs and CD34+ cells: After 5 days of treatment, the number of WBCs in the treated rats increased more than 2.7-fold compared with control group [(27.60±2.45) ×109 L-1, (10.11±1.81) ×109 L-1, P ＜ 0.01], CD34+ cells increased more than 12.2-fold compared with control group (38.31×107 L-1, 3.14×107 L-1, P ＜ 0.01). ② The extent of reendothelialization: At 14 and 28 days after balloon injury,carotid artery of reendothelialization in the treated group were (68.3±8.3)％ and (97.6±4.1)％, superior than the control group (33.8±6.3)％ and (76.1±5.2)％ (P ＜ 0.01). ③ The extent of neointimal hyperplasia: At 14 and 28 days after balloon injury, the neointima-media (I/M) ratios in the treated rats were 0.39±0.11 and 0.45±0.09, less than the control group 0.87±0.15,1.26±0.16 (P ＜ 0.01). A highly significant inhibition of neointimal hyperplasia was observed in the treated group. ④ CD31+ and vWF+ endothelial cells: At 28 days after injury, sections from G-CSF treated group showed almost complete and continuous monolayer of CD31 and vWF positive cells.In contrast, a patchy and interrupted CD31 and vWF positive cells were found lining the lumen of control group.CONCLUSION:Treatment with G-CSF significantly increases the number of CD34+ cells and accelerates the rate of reendothelialization of injured vessels, leading to marked inhibition of neointimal formation after vascular injury in rats.

Objective To evaluate the therapeutic effects of ultrasound biomicroscopy (UBM) assisted intraocular lens suspension surgery with one loop suturing for residual lens capsule aphakia.Methods Fifty aphakia patients (50 eyes) with residual lens capsular equal or greater than 180 degrees were chosen.The patients were randomly divided into 2 groups and underwent intraocular lens suspension surgery with one loop suturing:Trial group (25 cases,25 eyes) selected suture site according to the preoperative UBM results,and control group (25 cases,25 eyes) selected the suture site according to the clinical experience.The uncorrected visual acuity (UCVA),astigmatism at preoperative and postoperative 1 week,1 month and 3 months were observed,the eccentric and tilt value of the lens were calculated by UBM measurement at 3 month after operation,and the postoperative complications of the two groups were observed.Results UCVA at each time point after operation were higher than that before operation,and gradually increased with time.There was no significant difference between the two groups at 1 week before and after the operation of UCVA (P ＞ 0.05).At 1 month and 3 months after operation,UCVA in trial group were significantly higher than those of control group (all P ＜ 0.05).The astigmatism value at 1 week after operation in two groups was higher than that before operation and gradually decreased with time.The astigmatism value in trial group at 1 month and 3 months after operation had no significant difference with that of preoperative (P ＞ 0.05),while the astigmatism value in control group increased (all P ＜ 0.05).The astigmatism values had no significant difference between the two groups before and 1 week after operation (all P ＞0.05),which were significantly lower in trial group at 1 month and 3 months after operation (all P ＜ 0.05).Three months after surgery,the proportion of lens accurately implanted in the ciliary sulcus was significantly higher than control group (P ＜ 0.01),the eccentric and tilt values of the lens in trial group were significantly less than control group (P ＜ 0.01).No serious complication was observed in all patients.Conclusion UBM assisted intraocular lens suspension surgery with one loop suturing can significantly improve the postoperative visual quality,safety and lens stability,which can be used as a routine preoperative examination of intraocular lens suspension surgery.In addition,UBM provides an alternative method for the calculation of the eccentricity and tile value of the intraocular lens after the implantation of the intraocular lens.

Objective To study the correlation of 5 polymorphisms of Han Chinese patients in Sichuan Province with age-related macular degeneration (AMD). Methods The blood samples from 384 Han Chinese patients diagnosed with AMD and another 384 matched controls were collected using case-control study method. The chosen gene single nucleotide polymorphisms (SNPs) were genotyped by SnaPShot classify technology in the patients with AMD and 384 controls of Chinese Han population. Results All of the 5 genetype frequencies of the SNPs were in accordance with Hardy-Weinberg Equilibrium (P > 0.05). There were no statistically significantdifferences between the AMD group and the control group in the rs13117504 G allele frequency (P = 0.037, OR=1.24, 95%CI:1.01~1.53), the rs10033900 C allele frequency (P=0.023, OR=1.27, 95%CI: 1.03 ~1.57) and the rs1003390 frequency in the AMD dominant model (P = 0.039, OR = 0.74, 95%CI: 0.55 ~ 0.99). There were no statistically significant differences between the groups in the rs6822976 A allele frequencies (P =0.158), the rs7438961 G allele frequencies (P = 0.798) and, the rs7671905 T allele frequency (P = 0.909). The rs10033900 in the recessive model of AMD had no significant difference as compared to that in the control group (P = 0.107). The two groups showed no significant differences in both the dominant and recessive model of AMD in terms of the frequencies of rs13117504, rs6822976, rs7438961 and rs7671905 (P > 0.05). Conclusion The rs13117504 and rs10033900 of SNPs near CFI gene upstream has significant association with age-related macular degeneration , while the rs6822976 , rs7438961 , rs7671905 of SNPs have no significant correlations with age-related macular degeneration in Han Chinese population.

Medical cell biology is a very important course for medical international students. It is a new challenge for medical colleges to carry out the international students curricula in English. In order to successfully accomplish the teaching of cell biology course ,the teachers in Teaching and Research Section of Medical Cell Biology and Genetics in Guangzhou Medical University made the following preparation before class:choosing or compiling English teaching material,writing English teaching plan,making English PPT and trial teaching before lass. Meanwhile,they improved the teaching links in theoretical teaching and experimental teaching and emphasized on students' feed-back,which provided references for teaching of medical cell biology course.

Objective To investigate long-term data of dry eye after LASIK in young myopia patients.Methods Young myopia patients were divided into treated group (group A) and control (group B),who were interviewed before surgery and on 1 year,3 years and 5 years after surgery.Their diopters,uncorrected visual acuity (UCVA),tear breakup time test,Schirmer I test and corneal fluorescent staining were taken,then dry eye was diagnosed and OSDI questionnaire were scored.Results The postoperative incidence of dry eye in group A was higher than that before surgery (35.6％) and decreased with time (51.7％,44.9％ and 40.7％,respectively),and there was no significant difference in group B at each time point (P ＞ 0.05).The preoperative incidence of dry eye was equal between two groups (P ＞ 0.05),which were much higher in group A after surgery (all P ＜ 0.05).In group A,the postoperative constituent ratio of mild dry eye was the majority and decreased with time (65.6％,52.8％,41.7％,respectively),while moderate and severe dry eye was increased,especially the latter (6.6％,11.3％ and 16.7％,respectively;P ＜ 0.05).At 5 years after surgery,the UCVA and visual function score in dry eye patients were lower than those of non-dry eye patients (all P ＜ 0.05),while the residual refraction and ocular symptom score were higher than those of dry eye patients (all P ＜ 0.05).Conclusion Dry eye disease may last for a long time and may getting worse after LASIK.Therefore,we should pay more attention to the development of dry eye long after LASIK in young myopic patients.

Objective To investigate the effects of low frequency pulsed magnetic fields (LF-PMFs) on proliferation, the cell cycle, apoptosis, migration, cytoskeleton formation and nitric oxide (NO) secretion in cardiac microvascular endothelial cells (CMECs). Methods LF-PMFs with rectangular and triangular waveforms at 15 Hz were tested. The CMECs were randomly divided into a control group, 0.6 mT group, 1.2 mT group, 1.8 mT group and 2.4 mT group. Except for the control group, the CMECs were exposed to LF-PMFs 4 h/day for 5 days. Results After 5 days of intervention with rectangular wave LF-PMFs the proliferation of CMECs accelerated and NO secretion was enhanced significantly. The percentages of cells at ( S+G2 ) phase increased significantly, whereas apoptosis rates were significantly lower than in the control group. The migration of CMECs was facilitated, stress fibers increased and cytoskeleton components were reorganized. After 5 days of intervention with triangular wave LF-PMFs proliferation of CMECs accelerated and NO secretion was significantly enhanced. The percentages of cells at(S+G2)phase and migration increased, while apoptosis was inhibited. Cytoskeleton components were reorganized after exposure to 0.6 mT and 2.4 mT triangular waveform LF-PMFs. No significant change was detected with 1.2 mT and 1.8 mT triangular waveform LF-PMFs with regard to these variables. Conclusions The effects of LF-PMFs on proliferation, the cell cycle, apoptosis, migration, cytoskeleton formation and NO secretion function of CMECs were infiuenced by the waveform in addition to it's intensity and frequency.

Objective To investigate the risk factors of reversible posterior leukoencephalopathy syndrome (RPLS) in pre-eclampsia or eclampsia gravida. Methods This study was conducted in the Third Affiliated Hospital of Guangzhou Medical University between January 2013 and March 2016. A total of 100 patients who had no severe neurological diseases and were diagnosed pre-eclampsia or eclampsia, and underwent brain MRI were collected retrospectively. They were divided into 2 groups according to the MRI results, the RPLS group (n=49) and the non-RPLS group (n=51). The medical history, clinical symptoms and the results of laboratory examination were analyzed by the logistic regression, in order to explore the risk factors.Results In single factor analysis, HELLP syndrome, pregnancy associated with other diseases, poor prenatal care, grade 3 hypertension, elevated systolic blood pressure or diastolic blood pressure, elevated WBC, aspartate transaminase (AST), alanine aminotransferase (ALT), uric acid (UA) and lactate dehydrogenase (LDH), decreased platelet (PLT), headache, visual changes, seizures and conscious disturbance were more frequent in the RPLS group than those in the non-RPLS group (all P<0.05). According to the multivariate logistic regression analysis, the elevated WBC (OR=1.291, 95%CI:1.058-1.575, P=0.012), UA (OR=1.008,95%CI:1.001-1.016,P=0.032) and headache (OR=18.260, 95%CI:3.562- 93.607, P=0.000) were the independent risk factors.Conclusions Maternal history, clinical symptoms and some laboratory examinations might help in the early diagnosis of RPLS in pre-eclampsia or eclampsia gravida. Headache, the elevation of WBC and UA were the most significant factors.

Objective To investigate the role of early detecting macrophage inflammatory protein‐1β(MIP‐1β) and proealcitonin (PCT ) level for the diagnosis of spontaneous bacterial peritonitis (SBP) in the decompensated stage of liver cirrhosis .Methods 384 cases of decompensated stage of liver cirrhosis complicating SBP collected in the Affiliated 3201 Hospital of Xi ′an Jiaotong Univer‐sity from May 2011 to February 2015 were included into the SBP group ,while other 377 cases of decompensated stage of liver cir‐rhosis complicating ascites were included into the control group .The serum and ascites samples were collected for detecting PCT by using electrochemical luminescence method and MIP‐1β by using the enzyme‐linked immunoassay .The significance of these two in‐dicators was compared between the serum detection and ascites detection .At the same time the clinical application value of these two indicators was analyzed by using the receiver operating characteristic curve .Results The serum and ascites PCT and MIP‐1βlevels in the SBP group were significantly higher than those in the control group ,the difference was statistically significant (P<0 .05) ;the serum PCT level in the SBP group had statistical difference between the patients with Gram‐negative bacteria infection and the patients with Gram positive bacteria infection (P< 0 .05) ;the ascites MIP‐1β level in the patients with Gram‐negative bacte‐ria infection of the SBP group was higher than that with Gram positive bacteria infection ,the difference was statistically significant (P< 0 .05) .Conclusion The serum and ascites PCT and MIP‐1β detection can help to the differentiation diagnosis of early decom ‐pensated stage of liver cirrhosis complicating SBP ;the serum PCT detection is superior to the MIP‐1β detection ,while ascites MIP‐1β detection is superior to the PCT detection .

Objective To construct a recombinant human adenovirus type 3 ( HAd3 ) vector ex-pressing one major epitope of dengue virus type 1.Methods The gene encoding the envelope protein (304-314 aa) of dengue virus type 1 was inserted into the hypervariable region 1 ( HVR1 ) of HAd3 hexon by using overlap PCR.The recombinant gene was cloned into the shuttle plasmid, then linearized with AsisⅠrestriction enzyme and co-transformed into Escherichia coli BJ5183 strains with the digested backbone plas-mid for homologous recombination.The recombinant plasmid pBRAdΔE3GFP-DENV1 was transfected into AD293 cells to rescue recombinant adenovirus strains (rAdΔE3GFP-DENV1).ELISA and Western blot as-say were performed to evaluate the humoral responses induced in BALB/c mice after the immunization with rAdΔE3GFP-DENV1 strains.Results The recombinant adenovirus strains were successfully rescued. ELISA and Western blot assay showed that the antibodies in serum sample could recognize dengue virus type 1 strains.Conclusion The recombinant adenovirus strains expressing the epitope of dengue virus type 1 were successfully constructed.This study provided evidence for the development of multivalent vaccines against dengue virus.