OBJECTIVE: To explore the statistical methods for proficiency testing in the drug control field. METHODS: The statistical methods were sorted out with data processing used in inter-laboratory and proficiency testing by various industries both at home and abroad. The Grubbs test and Dixon test in classical statistical method were focused on as well as quartile method and iterative method in the robust statistical method, and the characteristics of the different statistic methods was summarized respectively. RESULTS: Every method has its advantages and application conditions. CONCLUSION: It is suggested that proper statistical method be chosen according to the data characteristics of the test results.

Objective To investigate the molecular mechanism of calreticulin ( CRT) transcription induced by HBV and its viral proteins. Methods The human hepatocellular cell line, HepG2, was trans-fected with pHBV1. 3 and eukaryotic expression plasmids of HBV viral proteins, respectively. The expres-sion of CRT was measured after transfection. A reporter plasmid of CRT promoter was constructed to analyze the induction of CRT promoter by pHBV1. 3 and HBV viral proteins. Furthermore, two truncated and one C/EBPα site deficient mutants were constructed to evaluate the regulatory effects of HBx on CRT promoter. Fi-nally, HepG2 cells were transfected with HBx expression plasmids and the cellular localization of C/EBPαwas analyzed. Results In this study, pHBV1. 3 could significantly up-regulate the expression of CRT at mRNA and protein levels as well as enhancing the activity of CRT promoter. Among the seven HBV viral proteins, HBx could enhance the activity of CRT promoter and the expression of CRT at mRNA and protein levels. HBx could not induce the transcription of CRT when the C/EBPα binding site was deleted from the CRT promoter. The expression of HBx could promote the nuclear translocation of C/EBPα. Conclusion HBV and its viral protein HBx could up-regulate the CRT expression at transcriptional level. The transcrip-tional factor C/EBPα played a critical role in HBx-induced transcriptional activation of CRT.

Objective To explore the effects of early growth response gene-1 (Egr-1) on bone marrow mesenchymal stem cells (BMSC) proliferation and osteogenic differentiation,which is aimed at providing new molecular targets for the treatment of osteoporosis.Methods Bone marrow was collected from adult men and the BMSCs were cultured primarily and observed by microscope.Meanwhile,flow cytometry was used for BMSCs phenotypic identification;After transfection of pcDNA3.1/Egr-1 into BM SCs,the level of BMSCs proliferation was determined by MTT respectively on the 2 d,4 d and 6 d;On the 7 d after transfection,the ALP activity assay was used for testing the ALP activity in BMSCs.And then,alizarin red S-calcium kit was used for measuring the calcified knots respectively on the 7 d,14 d and 21 d;On the 21 d after transfection,real-time qPCR and Western blotting were used respectively for measuring the expression of mRNA and protein of Egr-1,Runx2 and NDRG1;Further,BMSCs were transfected with Egr-1 siRNA,and the content of calcium nodules,ALP activity,the expression of Egr-1,Runx2 and NDRG1 were detected as above methods.Results The cells cultured in vitro showed high level of CD90 and CD29 and very low level of CD34 and CD45,which is accorded with the characteristic of BMSCs.The pcDNA3.1/Egr-1 transfection for BMSCs had no effect on cells prolifera tion.However,the calcified knots,ALP activity and the expression of Egr 1,Runx2 and NDRG1 were increased after transfection of pcDNA3.1/Egr-1 for BMSCs.In addition,Egr-1 siRNA showed the opposite effect with pcDNA3.1/Egr-1 transfection for BMSCs.Conclusion Egr-1 induces osteogenic differentiation of BMSCs by promoting NDRG1 but has no effects on proliferation of BMSCs.

Objectives To explore the clinical features and diagnosis of LMNA-associated congenital muscular dystrophy. Methods The clinical data from a case of muscular dystrophy caused by LMNA gene mutation were retrospectively analyzed. The related literatures were reviewed. Results A 8-month-old female infant suffered from weakness of raising head, eyelid droop, and motor development retardtion. LMNA gene was sequenced for the infant, her parents and the older sister. Heterozygous mutation of c. 94_96 del AAG (p. K 32 del) was found in the infant leading to the diagnosis of LMNA- associated congenital muscular dystrophy. No mutation was found in the infant’s parents and her older sister. The literature review showed that all ofLMNA- associated congenital muscular dystrophy patients had LMNA gene mutation, more than 80% patients mainly presented with weakness of raising head and may accompany with weakness of proximal limb, motor development retardation, and weakness of axial muscle. Conclusions Mutation analysis of LMNA gene is conducive to the diagnosis of congenital muscular dystrophy.

Objective Bone marrow mesenchymal stem cells ( BMSCs) , a kind of stem cells with multiple differentiation po-tentials, exist in the bone marrow and other organizations.This study aimed to investigate the repairing effect of the exogenous basic fi-broblast growth factor ( bFGF) against chronic obstructive pulmonary disease ( COPD) and its action mechanism, and to determine the expression of the bFGF gene in transfected rat BMSCs. Methods BMSCs were isolated, cultured and identified.The recombinant plasmid bFGF-pcDNA3.1 was constructed and sequenced.Liposome-mediated bFGF-pcDNA3.1 plasmid was transfected into the BM-SCs of the rat (bFGF-pcDNA3.1 transfection group), liposome-mediated pcDNA3.1 transfected into the BMSCs (pcDNA3.1 transfec-tion group) , and untransfected BMSCs used as the control.G418 screening was performed for 14 days.The gene and protein expres-sions of bFGF were determined by qRT-PCR and Western blot. Results The full-length sequence of the bFGF gene was consistent with that of the GenBank.The expression of the bFGF gene was significantly higher in the bFGF-pcDNA3.1 transfection group (7.028 ±0.568) than in the pcDNA3.1 transfection group (1.000 ±0.082) and the non-transfection control (1) (P<0.01), but with no statistically significant difference between the latter two groups (P>0.05).The expression of the bFGF protein was also re-markably higher in the bFGF-pcDNA3.1 transfection group (1.017 ±0.054) than in the pcDNA3.1 transfection group (0.217 ± 0.009) and the non-transfection control (0.165 ±0.013) (P<0.05), with no statistically significant difference between the latter two groups (P>0.05). Conclusion Mediated by the liposome reagent, the recombinant eukaryotic expression vector bFGF-pcD-NA3.1 can be transfected into rat BMSCs and expresses the bFGF gene and protein.

Objective To investigate the clinical features and genetic basis of cryopyrin-associated periodic syndrome (CAPS). Methods The clinical manifestations, laboratory tests, and genetic tests of one case of CAPS were retrospectively analyzed. The related literatures were reviewed. Results A 7 year and eight month old male patient had recurrent fever for 7 years and his whole body was covered with patchy red rash which was itchy and faded with pressure. The limbs and joints were normal. The levels of high-sensitivity C-reactive protein, erythrocyte sedimentation rate, rheumatoid factors were increased. The patient had fundus arteriosclerosis, double conjunctival lesions and nerve deafness on both sides. There was no mutation found in NLRP3 gene coding region, but a heterozygous mutation (-2667G>T) had been found in 5 ' untranslated region. Compared with normal control, the mRNA level of NLRP3 increased 4.2 times and the expressions of IL-1βand IL-18 gene increased 2.2 (P=0.002) and 1.2 times (P>0.05). Conclusions The clinical features of CAPS can be recurrent fever, rash, and joint involved. The oph-thalmologic abnormalities and varying degrees of deafness may occur during the progression. The test of NLRP3 gene may help diagnosis.

A case of eosinophilic lymphoid granuloma(ELG)was reported and the related literatures were reviewed.ELG is rare in clinic. The etiology and pathogenesis of ELG was unclear.The clinical feature includes enlarged lymph nodes which were always predilected for the head and neck regions,eosinophilic granulocytes and serum IgE rising.Lymphoid tissue hyperplasis formation of lymphoid follicles with active germinal centres are common in pathological examination.There is diffuse infiltration of eosinophils in interfollicular and perivascular zones. Surgery,drug therapy and radiotherapy are all effective for the treatment,but recurrence is often.

AIM:To investigate the expression and the significance of VEGF-C/D in rat cornea after alkali burning as well as the role of lymphangiogenesis in the high-risk corneal transplantation rejection.
●METHODS:The model of alkali burn corneal was made. Different times corneas were taken to electron microscope for vascularization, and examined the expression of VEGF-C/D and VEGFR-3 in l, 3, 5, 7, 14, 28d. The other rat cornea after alkali burn were divided into four parts to penetrate keratoplasty, containing only blood vessels in the cornea ( group A ) , angiogenesis and lymphangiogenesis ( group B ) , lymphangiogenesis degenerating period ( group C ) , angiogenesis degenerating period ( group D ) . ln addition, there are also normal groups ( group N ) to compare the Rl values and survival time of corneal graft.
●RESULTS: Electron microscopy showed that, when the first 7d rat cornea appeared neovascularization after alkali burn, but not lymphangiogenesis. The occurrence of new blood vessels and lymphatic in 2wk. There were no obvious lymphangiogenesis in 5wk and the angiogenesis gradually subside in 8 wk. The expression of VEGF-C/D and VEGFR-3 in the corneas of rats were up-regulated in the third days after the injury, and reached its peaks at 5d. The average survival time of group N, A, B, C, D were (14.25±0.62)d, (9.35±1.02)d, (5.06±1.13)d, (8.71±0.83) d, (9. 44±1. 05)d after transplant cornea. Compared to the rest of the group, group B plant average survival time significantly shortened (P<0. 05), while compared with group B, the survival time of A, C, D groups were significantly longer (P<0. 05).
● CONCLUSION: VEGF - C/D and VEGFR - 3 are expressed significantly after corneal alkali burn. New lymphatic vessels can accelerate high - risk corneal transplantation immune rejection.

Objective To evaluate the biochemical function of rat nature three-dimensional acellular pancreatic bioscaffold (3D-APB) in promoting cell proliferation and differentiation in vitro.Methods The fresh pancreas from 10 rats were perfused to prepare 3D-APB.The biocompatibility of 3D-APB was eva luated.The experiment was divided into 4 groups based on 4 kinds of three-dimensional media for AR42J culture,including blank control group,ECM group,PLGA group and 3D-APB group.We compared the proliferation and differentiation of AR42J pancreatic acinar cell at 3 days,5 days,7 days and 10 days after seeding among 4 groups.Results The 3D-APB could represent a biocompatible scaffold capable of integrating within host tissue.The proliferation rate of AR42J cells by MTT in 3D-APB was higher,while the apoptosis rate by flow cytometry was lower than those in other 3 media,which were all significantly different (P ＜ 0.05),respectively.The protein expression of PDX-1 and PTF-1 by western blot in 3D-APB group was greatly higher than those in other 3 groups (both P ＜ 0.05).The mRNA expression of PDX-1 and PTF-1 through qRT-PCR was significantly higher than those in other 3 groups (both P ＜ 0.05).Conclusion Compared with the commonly used chemical and natural scaffold at present,3D-APB could promote cell proliferation and differentiation,which was more appropriate for regenerative medicine.

Objective To evaluate the efficacy and safety of adefovir dipivoxil(ADV)in treating patients with hepatitis B e antigen(HBeAg)positive chronic hepatitis B.Methods In this randomized,double blind,placebo-controlled,multicenter trial,210 eligible patients with HBeAg positive chronic hepatitis B were recruited and randomized(randomization ratio was 2:1)receiving ADV 10 mg/d for 48 weeks(ADV+ADV group,n=142)or placebo for 24 weeks followed by ADV 10 mg/d for 24 weeks(PLB+ADV group,n=68).The primary endpoint was virological response. The secondary endpoint was serologic response(HBeAg loss rate and HBeAg seroconversion rate) and alanine aminotransferase normalization rate.Results After 24 weeks therapy,mean reduction of hepatitis B virus(HBV)DNA level comparing with that of baseline was 3.12 log_(10)copy/mL in ADV +ADV group while it was 0.95 log_(10)copy/mL in PLB+ADV group.The percentages of patients with HBV DNA clearance(HBV DNA level