The histological and biomechanical aspects of the osteogenesis in metaphysioto-my lengthening of the tibia were observed in 45 immature rabbits,which were divided into 3 groups as follows:(1)Group A:After metaphysiotomy,the periosteum and endosteum of the transected boneends were well-preserved.(2)Group B:After metaphysiotomy,the periosteum about 0.7 cm in width on the 2 bone ends was circumcised.Then the 2 bone ends were aligned without disturbing the endosteum.(3)Group C:After metaphysiotomy,a part of the bone marrow and endosteum in the 2 bone ends wwere removed.Then the 2 bone edns were well aligned and the periosteum was sutured up.In all the experimental animals,the tibia was lengthened by 1mm/day.It was found that in Group A,the formation and remodelling of the new bone in the lengthened area occurred quite early after metaphysiotomy and the new bone was good in its biomechanical quality;in Group B,the formation of new bone was retarded and the biomechanical quality of the new bone was not satisfactory;and in Group C,new bone formation was sluggish in the early stage after metaphysiotomy but it wwas active in the late stage,and the neww bone formed was similar to that of Group A in its histological morphology and biomechanical quality.The periosteum could grow synchronously with the slow traction without any tearing.In addition,osteotomy and mechanical traction could stimulate and accelerate periosteal osteogenesis.It is believed that an intact periosteal cuff is of significance in the osteogenesis of the lengthened area since it serves as a restriction framework for the adherent growing of the connection between the new bones of the 2 bone ends and periosteal osteogenesis plays a more important role than endosteal osteogenesis in the bony healing of the lengthened area.

Objective To establish the technique of biphasic seeding of articular chondrocyte s in vitro for cartilage engineering in u sing a self designed biological gel a nd the three-dimensional scaffold a nd observe the efficiency of cartilage regeneration in vitro-tissue engin eered articular cartilage from cell scaffold complex.Methods The articular chondrocytes were iso lated enzymatically from the epiphyseal cartilage of young rabbits,and were then plated i nto the tissue culture flasks and were cultivated.The first passage ar-ticular chondrocytes were collecte d and mixed fully with the self-made l iquid biological gel-matrix at appr oxi-mately 2.5?10 7 cells /ml to form cell-gel fluid.The cell-gel fluid was dropped into the p orous CPPf /PLLA(calcium polyphosphate fiber /poly -L -lactic acid)scaffold,and a cell-gel-scaffold c omplex was formed after being solidified.The complexes were cultivated for 4weeks.The changes of complexes in morphology and synthesis of collagen typeⅡandⅠand aggregates were investigated under the gross and the phase and light microscope.The GAG sulfate content in complexes was quantitatively mea sured by the modified dimethyl-methylene blue method.Results1)After feeded,the porous CPPf /PLLA s caffolds were completely filled with the liquid chondrocyte-gel and the chondrocyte-gel-scaffold comp lexes were well formed by solidification.During the cultivation,the complex es could keep its original shape and m aintain the stable homogeneous three-dimensional distribution of chondrocytes in themselves without cell falling.In the same time,the com-plexes were gradually increasing th e consistency with the elasticity an d lubrication surface.2)The chondro-genesis began in the periphery area a nd extended to the central area of the complexes with the passage of cultivation period.After the 2nd we ek,the complexes were gradually reorganized into the mature engineered cartilage with typical cartilaginous histological structure,with ric h typeⅡcollagen and the strong typical het-erochromia to toluidine blue,but wi th gradually fading negative immunological stain of collagen typeⅠ.Meantime,the scaffold was graduall y de-graded in the complexes.The average content of GAG sulfate in the enginee red cartilages at 4th week was(15.70?2.00)mg /g(wet weight ),and was over 30%of the natural articular cartila ge.Conclusion The technique of chondrocytes biphasic seeding for three-dimensional scaffold has advantages of simple manipulation,fix ing cell stably in the scaffold,main-taining the original shape of the com plexes and the stable homogeneous th ree-dimensional distribution of chondrocytes in the scaffold,and ad vancing the regeneration and matura tion of tissue-engineered articula r cartilage.[

Objective To establish the methods of efficiently isolating chondrocytes from epiphyseal cartilage and observe the basic biological characteristics of the cultured ones. Methods The improved isolation method through three-step enzymatic digestion using 1 g/L trypsase in 1 g/L ethylenediaminetetraacetic acid, 1 g/L hyaluronidase and 2 g/L type Ⅰ collagenase was established to isolate the primary passage chondrocytes from epiphyseal cartilage of young rabbits and its efficiency evaluated. The cellular biological characteristics of the chondrocytes were observed under the inverted and light microscopes. Results (1) All chondrocytes were isolated from the matrix after it was completely digested by three above-mentioned enzymes. The harvested chondrocytes significantly and positively correlated with the wet weight of cartilage. The number of chondrocytes from 1 g of wet cartilage was 32.3?106 with a viability of 97.6% on average. (2) The chondrocytes of the primary and the first passages showed triangle or multi-angle shapes (oval during growth and fusion) with the positive immunohistochemical stain of collagen type Ⅱ and the strong heterochromia to toluidine blue. The chondrocytes after the third passage became spindle-shaped with the negative stain of collagen type Ⅱ and the weak heterochromia to toluidine blue. Conclusions (1) The chondrocyte-isolating method through three-step enzymatic digestion has advantages of high efficiency in harvesting primary chondrocytes, high cellular viability and simple manipulation. (2) The chondrocytes differentiate and lose their special phenotypes gradually with passage.

Objective To establish the measurement for evaluating the efficiency of isolating and harvesting marrow mesenchymal stem cells (MSCs) and observe the main biological characteristic of MSCs in vitro during the passage cultivation. Methods ①The marrow nucleated cells in young rabbit marrow were isolated by the gradient centrifugation and cultivated to isolate and harvest MSCs. The efficiency of isolating and harvesting MSCs was measured by the number of the first passage MSCs from every 10~(6) of the marrow nucleated cells. ②The changes of MSCs morphology, growth and proliferation, synthesis of collagen typeⅠand activity of alkaline phosphatase (ALP) were observed during the passage cultivation. Results ① After cultivated at 2.5?10~(5)/cm~(2) marrow nucleated cells for 9-10 d, the first passage MSCs were harvested. The number of the first passage MSCs had a remarkable statistical positive correlation with the number of the marrow nucleated cells from one rabbit(r=0.932, P

Objective To investigate the adaptation of metabolism of the arthrodial cartilage for limb lengthening. Methods Thirty New Zealand rabbits were divided into two groups: limb lengthening group (A) and delayed observation group (B). Each group was further divided into three sub groups ( n =10) according to the distraction ranges: 10%, 20%, and 40%. Upper tibia metaphysis lengthening was adopted at 1 mm/d rate of distraction and iconography changes of the knee joint were observed with X ray. The contents of glycosaminoglycan (GAG), hydroxylysine (HYL), and hydroxyproline (HYP) were determined by biochemical techniques. Results ① There was no obvious imaging change of knee articular cartilage at every time point in each group. ② The content of GAG was normal in group A 10 , and B 10 , and B 20 , but the content of galactosamine decreased obviously in group A 20 ( P 0.05). The content of GAG recovered obviously in group B 40 as compared with that in group A 40 . ③ The ratio of HYL to HYP was 0.18 and 0.17, respectively in group A 40 and group B 40 , but it was normal in other groups, nearly 0.24. Conclusion Metabolism of the arthrodial cartilage during low distraction range can adapt the stimulation of distraction, but great distraction range may result in transitory disorder in metabolic function of the arthrodial cartilage.

Objective To observe the effect of isolating and harvesting the chondrocytes from rabbits rib cartilage with the method of three-step enzymatic digestion, and the biological characteristic of the isolated chondrocytes during cultivation in vitro to evaluate their biological activity. Methods The method of three-step enzymatic digestion was designed that the rib cartilage was digested one by one with 1 g/L trypsin and 1 g/L EDTA, 1 g/L hyaluronidase and 2 g/L collagenaseⅠ in the culture medium to isolate chondrocytes. The harvesting and viability rate of the primary chondrocytes were detected. During the passage cultivation in vitro, the changes of the chondrocyte shape and growth were observed, and the changes of the collagen typeⅠ and Ⅱ and aggrecan in the extracellular matrix were detected. Results ① The extracellular matrix of rib cartilage was completely dissolved by the three-step enzymatic digestion, and the chondrocytes were completely isolated from the solid matrix. The number of the harvested chondrocytes from every gram of wet cartilage was (4 295.7)?10~(4) on average,and their viability rate was 97.2% on average. ②The primary and first passage chondrocytes had triangle or multi-angle shape, and became elliptic shape at the growing confluence with the positive immunohistochemical staining of collagen type Ⅱ and the strong heterochromia to toluidine blue. The content of sulfate glycosaminoglycans(GAG) in the extracellular matrix of the primary passage cells was (80.61?11.40) ?g/cm~(2). The chondrocytes after the third passage gradually became spindle shape with the negative staining of collagen typeⅡ and the weak heterochromia to toluidine blue. The content of sulfate GAG of the fourth passage cells was (44.74?10.18) ?g/cm~(2). Conclusion ① The method of three-step enzymatic digestion can make the extracellular matix of rib cartilage to be completely degraded, and has advantages of the high efficiency of harvesting primary chondrocytes with high cellular viability rate and simple manipulation. ②The primary and first passage chondrocytes have fine biological activity.

Objective: To investigate the occupational health status of medical radiation workers in Hangzhou City, so as to provide the basis for their occupational health risk assessment. Methods: Data on medical radiological workers who underwent occupational health examinations from 2021 to 2022 were collected through the Physical Examination Information Management System of the Hangzhou Occupational Disease Prevention and Control Hospital. The physical examination data including blood routine, eye lens, thyroid ultrasound, thyroid function, liver function, renal function and blood lipid were collected, and the abnormal rates of occupational health examinations among workers with different genders, working years and occupational exposure types were analyzed. Results: A total of 3 968 medical radiation workers were investigated, including 2 310 males (58.22%) and 1 658 females (41.78%). There were 2 039 (51.39%), 821 (20.69%) and 1 108 (27.92%) workers with 1-<6, 6-<10 years and 10 years and above of work, respectively. Diagnostic radiology was the predomenant type of exposure, with 2 240 workers accounting for 56.45%. The abnormal rates of thyroid ultrasound and blood lipid were 47.73% and 45.21%, respectively, which were relatively higher than other items. The abnormal rates of micronucleus rate, thyroid ultrasound, thyroid function and renal function were higher in females than in males, while the abnormal rates of lymphocyte count, liver function and blood lipid in males were higher in males than in females (all P<0.05). With the increase of working years, the abnormal rates of micronucleus rate and blood lipid showed upward trends (both P<0.05). There were statistically significant differences in the abnormal rates of thyroid ultrasound, liver function and blood lipid among different occupational exposure types (all P<0.05). Conclusion Long-term low-dose ionizing radiation environment affects the thyroid, micronucleus rate and blood lipid of medical radiation workers in Hangzhou City, with differences observed among workers with different genders and occupational exposure types.

Objective To conduct the amplification,cloning,bioinformatics analysis,prokaryotic expression and purification of enterovirus 71 VP1 gene segment and to initially confirm the biological activity of the recombinant expression product.Methods A pair of specific primers was designed according to GenBank EV71 sequence,viral RNA as a template was extracted from the throat swab specimens in the EV71 patients.EV71 VP1 gene was amplified by RT-PCR.After enzyme digestion,the expression vector pET28a was inserted.The prokaryotic expression vector of pET28a-EV71 VP1 was constructed.Then the E.coli DH5a transforma-tion was performed.IPTG was adopted for induction expression.The expression results were analyzed by using SDS-PAGE and Western blot.The bioinformatics analysis of the sequenced results was performed by the software.Expressed protein was purified and the plates were coated,ELISA was used to test the VP1 specific IgG antibody in serum samples of EV71 positive and COX A16-positive patients.Results The BLAST alignment showed that the homology of the objective gene EV71 VP1 was 99% com-pared with other strains(JQ766207.1)in GenBank.EV71 VP1 protein was about 32×103 ,which mainly existed in the form of in-clusion body.The bioinformatics analysis showed that EV71 VP1 protein was a hydrophilic protein,without transmembrane region and N-terminal signal peptide sequence,the tertiary structure existed.The ELISA results showed that the specific IgG OD value in EV71-positive patients was(2.425±0.521),OD value in COX A16 positive patients was(1.205 ±0.314),the normal control OD value was(0.353±0.128).The sensitivity and specificity of EV71 VP1 protein detection were 84% and 88% respectively.Conclu-sion The pET28a-EV71 VP1 expression vector is successfully constructed;the preliminary analysis on the serum of the infected patients by ELISA shows that the obtained objective protein has higher sensitivity and specificity,which is initially confirmed to have biological activity and can be further used for the related study on EV71 diagnosis and vaccine.

Objective Psoriasis vulgaris (PV) is easy to prone to recur and hard to cure and little research has been done on combined treatment on PV.The article was to study the clinical effects of total glucosides of paeony capsules (TGP) combined with acitretin and compound flumethasone on PV as well as the peripheral blood cytokine levels.Methods 126 patients with PV who visited our hospital from October 2015 to January 2017 were randomly divided into combined treatment group (63 cases) and control group (63 cases).Both groups were treated with oral acitretin and topical compound flumethasone, what's more, the compound flumethasone group received oral TGP treatment, 8 weeks for a course.The clinical therapeutic effects were evaluated by the levels of peripheral blood IL-17, IL-18, IL-23, TNF-α level, PASI score and percentage of total skin lesions before and after the treatment.Results After the treatment, the concentration of IL-17, IL-18, IL-23 significantly decreased(P<0.05), which was significantly less in combined treatment group compared with control group (IL-17 [61.18±8.91] vs [78.64±7.85], IL-18 [68.56±17.95] vs [79.49±18.64], IL-23 [70.13±12.16] vs [91.18±16.89] pg/ML)(P<0.05).Moreover, the TNF-α level, the PASI score and the percentage of total skin lesions significantly decreased in both groups after treatment(P<0.05), which was significantly less in combined treatment group compared with control group (TNF-α level [14.47±7.53] vs [23.49±8.12]ng/L, PASI score [4.09±1.29] vs [7.29±5.13], the percentage of total skin lesions [6.17±4.59]% vs [8.09±5.18]%) (P<0.05).Conclusion TGP combined with acitretin and compound flumethasone can significantly enhance the clinical therapeutic effects and effectively regulate the levels of the IL-17, IL-18, IL-23 and TNF-α level, which results in treating psoriasis vulgaris.

Objective To compare the effect of 0.9％ NaCl solution, Ringer solution, and acetate sodium potassium magnesium calcium glucose solution (ASPMCG solution) on internal environment in patients with septic shock receiving early fluid resuscitation.Methods A prospective randomized controlled trial was conducted. From June 2016 to January 2018, a total of 280 patients with septic shock admitted to intensive care unit (ICU) of Northern Jiangsu People's Hospital were enrolled. All eligible patients were randomly divided into 0.9％ NaCl solution group (group N), Ringer solution group (group L) and ASPMCG solution group (group P) with random number table. In all patients, catheters were placed in the right internal jugular vein or subclavian vein and were infused with 30 mL/kg of corresponding crystalloid within 3 hours after admission. According to fluid responsiveness, the rehydration rate and fluid volume were determined by the researcher within 6 hours. Other treatments were based on the 2012 Surviving Sepsis Campaign (SSC) guidelines during the study. In this study, 6-hour or 24-hour fluid volume and 7-day exogenous insulin use were recorded. The changes in arterial blood pH, base excess (BE), blood glucose (Glu), lactic acid (Lac), and serum Na+, K+, Cl-, Ca2+ were observed at 0, 3, 6, 24 hours, and 3 days and 7 days of resuscitation.Results In this study, a total of 1082 patients were admitted to the ICU, and patients who did not meet the diagnostic criteria for septic shock, death or discharge within 24 hours of ICU admission were excluded. Finally, 280 patients with septic shock were enrolled in the analysis, with 94 patients in group N, 94 patients in group L and 92 patients in group P. There was [Abstract] Objective To compare the effect of 0.9％ NaCl solution, Ringer solution, and acetate sodium potassium magnesium calcium glucose solution (ASPMCG solution) on internal environment in patients with septic shock receiving early fluid resuscitation.Methods A prospective randomized controlled trial was conducted. From June 2016 to January 2018, a total of 280 patients with septic shock admitted to intensive care unit (ICU) of Northern Jiangsu People's Hospital were enrolled. All eligible patients were randomly divided into 0.9％ NaCl solution group (group N), Ringer solution group (group L) and ASPMCG solution group (group P) with random number table. In all patients, catheters were placed in the right internal jugular vein or subclavian vein and were infused with 30 mL/kg of corresponding crystalloid within 3 hours after admission. According to fluid responsiveness, the rehydration rate and fluid volume were determined by the researcher within 6 hours. Other treatments were based on the 2012 Surviving Sepsis Campaign (SSC) guidelines during the study. In this study, 6-hour or 24-hour fluid volume and 7-day exogenous insulin use were recorded. The changes in arterial blood pH, base excess (BE), blood glucose (Glu), lactic acid (Lac), and serum Na+, K+, Cl-, Ca2+ were observed at 0, 3, 6, 24 hours, and 3 days and 7 days of resuscitation.Results In this study, a total of 1082 patients were admitted to the ICU, and patients who did not meet the diagnostic criteria for septic shock, death or discharge within 24 hours of ICU admission were excluded. Finally, 280 patients with septic shock were enrolled in the analysis, with 94 patients in group N, 94 patients in group L and 92 patients in group P. There was no significant difference in the amount of crystalloid, colloidal fluid, total fluid within 6 hours or 24 hours or exogenous insulin dose within 7 days among the three groups. After fluid resuscitation, blood Cl- concentration in the three groups was increased in different degrees, peaked at 24 hours, but it in group N was significantly higher than that in group L and group P (mmol/L: 107.5±5.6 vs. 106.1±4.8, 105.1±4.2, bothP ﹤ 0.05). Moreover, blood Ca2+concentration also showed an increased tendency, it was significantly lower at 3 hours and 6 hours in group N than that in group L and group P (mmol/L: 1.10±0.08 vs. 1.15±0.09, 1.12±0.10 at 3 hours, 1.12±0.10 vs. 1.16±0.08, 1.15±0.09 at 6 hours, all P < 0.05). There was no significant difference in blood Cl- or Ca2+between groups L and P (bothP > 0.05). Arterial blood pH, BE, Glu, Lac, Na+, or K+at each time point during fluid resuscitation also showed no difference among the three groups.Conclusion Among patients with septic shock receiving early fluid resuscitation, compared with Ringer solution and ASPMCG solution, 0.9％ NaCl solution may cause hyperchloremia and hypocalcemia, but has no significant effect on acid-base balance.Clinical Trial Registration Chinese Clinical Trial Registry, ChiCTR-IOR-16009176.