Objective To explore the changes and clinical significance of serum human cartilage glycoprotein-39(HC gP39),osteopontin (OPN) and rheumatoid factor (RF) in patients with rheumatoid arthritis (RA).Methods Serum HC gP39,OPN levels were measured by ELISA in 98 patients with RA and 98 healthy controls.Serum RF was detected by nephelometric immunoassay.Results The serum HC gP39,OPN and RF levels of RA group were significantly higher than the healthy control group (t =20.25,32.71,36.34,all P ＜ 0.01),and serum HC gP39,OPN and RF levels in active phase patients were higher than the inactive phase patients (t =24.22,45.62,50.15,all P ＜0.01).The levels of serum HC gP39,OPN increased gradually with the increase of the staging severity(F =18.48,12.36,all P ＜0.05).The serum level of RF was positively correlated with the levels of HC gP39,OPN in patients with RA (r =0.682,0.656,all P ＜ 0.01),the serum level of HC gP39 was positively correlated with the level of OPN (r =0.608,P ＜ 0.01).Conclusion The HC gP39,OPN and RF reflect situation of RA patients and have close relationship with the clinical progression of RA,which can be used as one of important indexes to judge RA activity and different staging.

To control and prevent the Echinococcus in the place ,we established a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Echinococcus species specific DNA from dog faeces .Four primers which recognizing 6 dis-tinct regions on the NADH dehydrogenase subunit 2 (ND2) gene of Echinococcus granulosus were designed and used for LAMP assay .The specificity of LAMP assay was evaluated using DNA extracted from Echinococcus granulosus , Taenia saginata , and other dog intestinal parasites .In addition ,the sensitivity of LAMP assay was compared with that of conventional PCR using recombinant plasmid carrying Echinococcus granulosus ND2 gene fragment as standard template DNA after 10-fold serial dilution .Furthermore ,we extracted DNA from 46 canine fecal samples collected from endemic areas ,and tested the copro-DNA samples using LAMP and necropsy method .Results showed that E .g ND2 primer sets could differentiate Echinococcus granulosus from Echinococcus multilocularis without cross reaction among other parasites detected .Furthermore ,the LAMP assay with primer sets to the ND2 gene could detect 4 × 101 copies of target gene ,demonstrating 103 times higher sensitivity than that of conventional PCR methods .The LAMP assay with primer set to ND2 gene showed good sensitivity and specificity to detect copro-DNA samples extracted from fecal samples of 46 dogs tested in endemic areas .There was no statistically signifi-cant difference among LAMP and necropsy .In this study ,a sensitive ,specific and rapid copro-DNA detection LAMP assay was developed successfully for diagnosis of dogs infected with Echinococcus granulosus .Due to its rapidity ,simplicity ,speci-ficity and sensitivity ,the LAMP assay is a promising new tool for rapid detection of dogs infected with Echinococcus spp .dur-ing the field survey or in poor-equipped laboratories .

Objective To comprehend etiology and clinical manifestation changes of infant pneumonia in this locality.Methods Indirect immunofluorescence (IIF) assay was applied in children with acute pneumonia to detect serum 11 kinds of viruses[respiratory syncytial virus (RSV),adenovirus (ADV),influenza virus (IFV-A+B),parain fluenza virus(PIV14) ,coxsackie B,virus(CB1V),Coxsackie A7 virus (CA7V) ,ECHO virus]specific antibody IgM,according to the serum virus-specific IgM positive,C-reactive protein(CRP)<8mg/L and no other pathogenic infection and laboratory evidence for the conditions of 436 cases detected in children with pneumonia.Results Detected a total 125 cases of antibody-positive,the positive detection rate is 37.99%.Of which 103 cases of single virus infection .accounting for 82.4% ,22 cases of mixed infection,accounting for 17.6%.RSV infection on top of the list followed by the rest of IFV,ADV and PIV.Infants of different ages,different seasons of the different types of virus susceptibility.Conclusion Pneumonia in infants were caused by pathogenic bacteria in addition to the virus of a wide range,and the incidence of age,the peak seasons and the clinical manifestations were vary.From an early stage of infection pathogen detection,clearing pathogen type,making the correct diagnosis of pneumonia in the treatment of infants had an important guiding significance.

Objective To investigate the relationship between the expression of β-glucuronidase mRNA in hepatocellular carcinoma(HCC)tissue and the clinicopathologic factors.Methods The expressions of β-glucuronidase mRNA in 25 samples of HCC tissue and 10 samples of normal hepatic tissue were detected by RT-PCR,and the relationship between β-glucurenidase mRNA in HCC tissue and clinicopathologic factors was analyzed.Results The expression of β-glucuronidase mRNA in HCC tissue was not influenced by the age of patient,tumor size and alpha-fetoprotein level.The expression of β-glueuronidase mRNA in patients with portal vein tumor thrombus or lymph node metastasis Was significantly higher than those without(t=7.857,9.341,P＜0.05).Conclusions The expression of β-glucuronidase mRNA is closely related to the existence of portal vein tumor thrombus and lymph node metastasis.β-glucuronidase may play a role in the invasion and metastasis of HCC.

To analyze the mutations of F12 genein one pedigree with congenital factor FXII (FXII) deficiency , and investigatethe molecular mechanisms of FXII deficiency . Methods Activated partial thromboplastin time(APTT),Prothrombin time(PT), FXII activity(FXII:C), FXII antigen(FXII:Ag) and other coagulant parameters were tested in the proband and his family members .5'and 3'UTR,all exons and their exon-intron boundaries of F12 gene were analyzed by direct sequencing .The detected mutations were confirmed by reverse sequencing .100 healthy persons were as normal controls .Results The proband showed a markedly prolonged APTT (106.4s), the FXII:C and FXII:Ag were 2.0% and 1.0%, respectively .Hissecond daughter and granddaughter had slightly prolonged APTT , and other family members are normal.The FXII:C and FXII:Ag of family members were also decreased ( his son, 23.0% and 21. 0%;his elder daughter , 23.0%and 23.0%;his second daughter ,24.0%and 23.0%;hisgranddaughter , 23.0%and 23.0%).The phenotype of all members is consistent with cross -reactive material negative . Nucleotide sequencing analysis showed that the proband had missense mutations in the F 12 gene, including one homozygous mutationc.1556T >G ( p.Leu519Arg) and a commonly reported single nucleotide polymorphism site within the promoter region of the F 12 gene (46T/T) .Sequencing results from the proband 'children demonstrate them as carriers of a heterozygous missense mutation .The proband 's wife is normal and with 46C/C in the promoter region .Conclusion The c.1556T>G in exon 13 is a novel mutation .This mutation affects FXIIcatalytic function , associated with a reduced level of FXII .

Objective To explore the treatment methods and prognosis of early infection and delayed infection after intramedullary nail fixation.Methods Data of 22 cases of postoperative infections after intramedullary nail from January 2013 to August 2017 were retrospectively analyzed.There were 18 males and 4 females aged from 20 to 72 years old,with an average age of 46.8 years.14 cases were tibias and 8 cases were femurs.In the early infection group,6 cases showed swelling,heat and pain in the affected area with drainage and pus.In the late infection group,12 cases showed sinus formation and 4 cases showed no sinus tract.According to whether the infection occurred within six weeks,it was divided into early infection and delayed infection groups.Of 6 patients in early infection group,there was 1 case of septic shock which underwent removal of intramedullary nails,debridement and antibiotic bone cement stick implantation.5 cases were retained intramedullary nail and underwent local debridement treatment.Late infection occurred in 16 patients.One patient with tibia infection was given partial dressing to heal the fracture.Then the intramedullary nail was removed and intramedullary debridement was performed.Two patients with poor general condition,the intramedullary nails were removed and debridement was performed.Calcium sulphate cement was implanted and fixed with external fixation.The remaining 13 cases were treated with debridement and antibiotic cement stick implantation.We compared the differences between early and late infections of internal fixation,infection control,fracture healing,and secondary fracture fixation.Results Of the 6 patients with early infection,1 patient with septic shock removed intramedullary nails to control infection.After infection controlled,the fracture was treated with intramedullary nailing.Of the 5 patients with retained intramedullary nails,2 patients' infection were controlled and 3 were uncontrolled.After removal of the intramedullary nails the infection was control.The success rate of retaining intramedullary nails was 33.3％ (2/6).Late infection occurred in 16 cases and infection was all controlled.The fractures healed in 22 patients.The fracture healing time of 6 patients with early infection was 2-6 months,with an average of 3.67±2.08 months.The fracture healing time of 16 patients with late infection was 2-4 months (average 3.2±0.79) months.Conclusion Patients with early bone infections after femoral and tibial intramedullary nail surgery may attempt debridement therapy with retained intramedullary nails,but the failure rate is high.If the intramedullary nail fails to remain,follow the treatment of patients with delayed bone infection.For patients with delayed bone infection,because the fracture has not yet healed,thorough debridement is used after the removal of internal fixation,then calcium sulfate or antibiotic bone cement stick should be implanted and fixed with external fixation.For the second phase,we may choose plate,intramedullary nail or external fixation to fix the fractures according to the soft tissue condition.All of the fixation methods could provide good fracture healing.

Objective:To investigate the efficacy of internal fixation maintenance after fracture-related infection (FRI).Methods:Retrospectively analyzed were the data of 81 patients with deep FRI after 6 weeks of internal fixation who had been treated with hardware maintenance at Department of Orthopedics, The Second Hospital Affiliated to School of Medicine, Zhejiang University between 2013 and 2021. They were 61 males and 20 females, aged from 11 to 73 years (average, 11 years). After admission, the patients received bacterial culture, thorough debridement, negative pressure suction, soft tissue repair, and local and intravenous antibiotics. If a joint was affected by FRI, its cavity was cleaned and drained. Infection control and fracture healing were regularly observed in all patients. A treatment was considered successful when the internal fixation was maintained until fracture union, and considered as unsuccessful when the internal fixation was removed before fracture union. Risk factors associated with treatment failure were identified from gender, age, smoking, diabetes, fracture type, methicillin-resistant Staphylococcus aureus (MRSA) infection, methicillin-susceptible staphylococcus (MSSA) infection, Pseudomonas aeruginosa infection, Escherichia coli infection, infection by two kinds of bacteria, negative bacterial culture, early infection (within 2 weeks) and local use of antibiotics.Results:All patients were followed up for an average of 30 months (from 6 to 84 months). Fracture union was achieved in 62 (76.5%) patients with infection control and internal fixation retained. Masquelet technique was used to treat bone defects in 2 patients; a muscle flap or skin flap was used to reconstruct soft tissue coverage in 11 cases; fracture union was achieved by antibiotics and dressing changes in 2 patients with sinus tract. Amputation was performed in one unsuccessful case due to uncontrollable infection, and internal fixation was changed to external fixation in the other 18 unsuccessful cases, of which 3 achieved final bone union after application of Masquelet technique, 7 achieved final bone union after application of bone transfer technique, and 3 achieved soft tissue coverage after reconstruction with flap technique. Pseudomonas aeruginosa infection, open fractures and FRI for more than 2 weeks were high risk factors for failure in internal fixation maintenance ( P<0.05). Conclusions:If internal fixation is still stable and effective, hardware maintenance should be tried first in the patients with FRI within 6 weeks after fracture internal fixation. Muscle flap or skin flap surgery should be performed as soon as possible to effectively control infection and promote fracture union in the patients with soft tissue defects after thorough and effective debridement. History of open fracture, Pseudomonas aeruginosa infection, and FRI for over 2 weeks may be risk factors for failure in internal fixation maintenance.