0.99). DC: reproducibility was good for neutrophils, lymphocytes monocytes, eosinophils and basophils. Comparison of the results by instrument with manual for normal samples in morphology, the correlation was better for neutrophils, lymphocytes and eosinophils (r:0.968～0.983) ,good for monocytes(r=0.917), not good for basophils(r=0.659);The WBC scattergram would change and alarm flags would display when there are neutrophilic stab granulocytes, abnormal or atypical lymphocytes and immature cells in higher proportion.Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to check with microscopy for DC before reported when it were doubtted.

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.

Objective To investigate the therapeutic effects of low-dose heparin on sepsis. Methods Seventy-nine sepsis patients were randomly divided into tow groups: beparin treatment group (n=37) and routine treatment group(n =42). The 7-day and 28-day mortality, the days in ICU and the length of stay, the changes of oxygenation index, the days of mechanical ventilation and the rates of disseminated intravascular coagulation (DIC), acute renal failure (ARF), acute respiratory distress syndrome (ARDS) and multiple organ dysfunction syndrome(MODS) were observed. The levels of APTT, PT and platelet (PLT) count were determined before and after treatment in two groups. Results The rates of DIC, ARF and MODS in beparin group decreased significantly after therapy: rate of BIC, 15.4% vs 38. 7% (P=0.03) ; rate of ARF, 25.0% vs51.9% (P=0.04); rate of MODS, 26.3% vs50.0% (P=0.04). In heparin group, the 28-day mortality was statistically reduced (15.4% vs 32.4%, P = 0. 03). The differences between beparin group and routine group were not statistically significant in the 7-day mortality (7. 7% vs 12. 9% ,P =0. 08) ,the days in ICU(Z =0. 281 ,P =0. 779,rank sum test) ,the length of stay (Z = 0. 562, P = 0. 574, rank sum test), the oxygenation index (P = 0. 82), the days of mechanical ventilation [(126.07±166.21)h vs (179.27±221.7)h,P=0.28] and the rate of ARDS (44.0% vs 46.2% ,P= 0. 88). The differences in APTT, PT and PLT were not significant between the two groups. Conclusion Low-dose beparin can decrease the mortality rate of sepsis and improve the prognosis of patients. It is a safe promising therapy in sepsis patients without severe side effects.

Objective The purposes of this study were to explore the relationship between personal-ity traits and coping strategies among primary glaucoma patients. Methods 100 patients with primary glaucoma participated the investigation adopting Eysenck Personality Questionnaire (EPQ) and the Medical Coping Methods Questionnaire (MCMQ). Data underwent analysis using descriptive and Spearman rank correlation methods. Results L(lie scale) was (57.83±4.13). The primary glaucoma patients usually took different coping style from norm, facing was (16.91±2.21), avoiding was (15.69±2.01), yielding was (11.54±2.03), all was significantly different than those of norm. L was evidendy negatively correlated with facing, positively correlated with avoiding. Conclusions The personality traits of primary glaucoma pa-tients are different from norm people. They usually take avoiding and yielding coping style, while facing coping style is rarely adopted. Patients with exocentric personality and easy to communicate are apt to adopt positive coping style, while in patients who are identical in emotion and lie, they may take avoiding and yielding coping styles.

Objective To investigate the clinical features and disease-causing mutations of familial hypomagnesaemia with hypercalciuria and nephrocalcinosis.Methods In February 2016,a 24 year old female patient with left kidney stone and nephrocalcinosis in bilateral kidneys was admitted to our hospital.One month prior to this admission,she had been treated by PCNL to remove the most part of left kidney stone in otherhospital.Mter admission,She was found hypomagnesaemia (serum magnesium 0.65 mmol/ L) and hypercalciuria (24h urine calcium 364.0 mg) but with normal renal function (serum creatinine 101.5μmol/L).And the remained part of left kidney stone was removed by flexible ureteroscope.As she was considered probably with an autosomal recessive FHHNC,an analysis of CLDN16 and CLDN19 gene mutations was performed using her and her parents'peripheral white blood cells.Results Mutation analysis revealed this patient had two heterozygous mutations in the CLDN16.One is an one-base deletion mutation in the 123th codon in exon 2:368delA.The other is a missense mutation in the 139th codon in exon 2:416C →T which resulted in an amino acid change Ala139Val.Her parents respectively had one of each heterozygous mutation.In the six months follow-up,an oral administration with hvdrochlorothiazide,potassium citrate,and calcium magesium supplements significantly reduced her hypomagnesaemia (serum magnesiun 1.0 mmol/L) and hypercalciuria (24-h urine calcium 156.0 mg),and no stone recurrence and aggravation of nephrocalcinosis and renal dysfunction occurred.Conclusions We diagnosed a patient with FHHNC who had a novel compound heterozygous mutation of CLDN16.This rare disease should be suspected if there are three constant clinical features of hypomagnesaemia,hypercalciuria and nephrocalcinosis,and verified with CLDN16 and CLDN19 gene test.Currently the option for treatment of FHHNC is symptomatic treatment until severe deterioration of renal function.The hydrochlorothiazide,potassium citrate,and calcium magesium supplements may have considerable effects on hypomagnesaemia and hypercalciuria.

Based on the conserved region nucleotide sequences of five potato viruses/viroid(Alfalfa Mosaic Virus,ALMV;Cucumber mosaic virus,CMV;Cucumber mosaic virus-satellite,CMV-sat;Potato virus Y,PVY;Potato spindle tuber viroid,PSTVd) and one inner control(18S rRNA),the microarray containing specific oligonucleotide probes and PCR probes were designed and fabricated.The effects of probe concentration,hybridization time,hybridization temperature and spotting solutions on microarray hybridiazation were evaluated.Finally the specificity of optimized plant virus detection array was validated.No significant effect on hybridization signal intensity was observed when the concentration of the oligonucleotide probes ranged from 5 to 20 ?mol/L,there was a linear relationship between the concentration of PCR probe and hybridization signal intensity.The greatest signal intensity were obtained when hybridized at 45℃ for 4 h,and the oligonucleotide probes and PCR probes had a similar effect on microarray hybrization.Among the different spotting solutions,DMSO produced a good reproducibility.The plant virus could be detected specifically by oligonucleotide probe microarray and PCR probe microarray after optimization.

Objective:To explore the relationship of teachers'job burnout with job stress,coping swategies associated with job stress,teachers'sense of efficacy,self-esteem,locus of control,social support in primary and secondary schools.Methods:Questionnaires about teachers'job burnout,job stress,coping strategies associated with job stress,teachers' sense of efficacy,self-esteem,locus of control,social support were applied to 766 primary and secondary teachers from Shandong and Hubei provinces.Results:(1)Teachers' job burnout was significantly positive correlated with its job stress,inactive coping and locus of control(r=0.23～0.40);(2)Teachers' job burnout was significantly negative correlated with its active coping,teachers' sense of efficacy,self-esteem and social support (r=-0.28～0.41).Conclusion:Teachers' sense of efficacy,job stress,self-esteem,locus of control,inactive coping, active coping and social support significantly can predict their job burnout.

Objective To explore a new method of functional reconstruction of hand digits and joints with free transfer of foot tissues so as to increase the success rate of the operation.Methods After micro-anatomic study of the plantar and dorsal metatarsal arteries,retrograde and free grafts of foot tissues pedicled with plantar metatarsal arteries were designed and applied in transplantation to treat 76 cases of hand digital or joint defects.The surgeries included 58 cases of transfer of the second toe,four cases of transfer of composite tissues of the second toe, eight cases of transfer of proximal interphalangeal joint,and six cases of nail flap transfer.Results The mi- cro-anatomic study found that the first plantar metatarsal artery was anatomically constant and the diameter of its branch to the second toe was larger than that of the first dorsal metatarsal artery.Flaps survived in 75 of the 76 patients(98.7%),with fine appearance and significantly improved function.One patient who had received free transfer of the second toe to reconstruct the thumb function had to undergo a second repair with infraclavicula skin tube because of refractory arteriospasm of anastomosed vessels.Conclusion Transfer with free retrograde grafts of foot tissues pedicled with plantar metatarsal artery to reconstruct hand functions can effectively improve the success rate of the operation,because it is free of the shortcomings of great anatomic variation of blood vessels and time-consuming and complex procedures in conventional transfer.

Objective : To explore the reform of the operating mechanism of centers for disease control and prevention ( CDCs ) in the new era, and to put forward recommendations. Methods : We collected the financial investment and staffing of CDCs in Zhejiang Province from 2018 to 2020 through China Disease Prevention and Control Information System and Zhejiang Health Statistical Yearbook. We conducted a questionnaire survey among 620 staffs selected by convenience sampling, and a qualitative interview among 40 leaders and experts from CDCs and general hospitals in Zhejiang Province. Results : By the year 2020, there were 105 CDCs in Zhejiang Province, including 1 province-level, 11 prefecture-level, 90 county-level and 3 development zone-level ones. The revenues were mainly from financial subsidy and business incomes, which were over expenditures. There were 7 008 posts in CDCs, with a staffing allocation ratio of 1.29/10 000. Among the reform measures of operation mechanism of CDCs, the flexible communication between clinicians and public health doctors was highly approved (90.03%); among the reform measures of enhancing the integration of prevention and treatment, further clarifying the public health functions of hospitals and ensuring financial compensation was highly approved (74.01%). Conclusion We should further stimulate the endogenous power and vitality of CDCs, strengthen the construction of talent team, promote the active integration of hospitals and CDCs, and improve public health level.

Objective To investigate the effects of different Leptospira strains on phagocytosis of peritoneal macrophages of guinea pigs,and explore the role of innate immune in the pathogenesis of leptospirosis. Methods Peritoneal macrophages of guinea pigs were infected in vitro by three different Leptospira strains,the virulent Leptospira interrogans serovar Lai type strain Lai,the avirulent L.interrogans serovar Lai type strain IPAV,and the nonpathogenic L.biflexa serovar Patoc type strain PatocⅠ,respectively,and heat inactivated Staphylococcus epidermidis was added 0.5,1.5,3 and 6 h after infection and incubated for 30 min.The effect of Leptospira on the phagocytosis of macrophage was evaluated by the inactivated Staphylococcus epidermidis phagocytosis rate and phagocytosis index.Phagocytosis and ultrastructure of peritoneal macrophages were observed by transmission electron microscopy 3 h after infection,and changes of cytoskeleton of the macrophages were observed by laser scanning confocal microscopy. Results The phagocytic rates and phagocytic indexes of strain Lai,strain IPAV and strain PatocⅠinfection groups were significantly lower than those of control group 3 h and 6 h after infection(P