BACKGROUND:Currently, bone graft is mainly used for repair of bone defects, and tricalcium phosphate is the most used artificial bone material. But the effectiveness of the tricalcium phosphate bone graft is stil controversial, and there is also no detailed report about its function during the healing of bone defect. ＜br> OBJECTIVE:To observe the concentration changes of bone morphogenetic protein-2 and vascular endothelial growth factor as wel as bone healing after tricalcium phosphate graft in bone defects. ＜br> METHODS:Forty-eight C57 mice were randomly divided to experimental group and control group. A 2-mm-long diaphyseal segment and periosteum from the middle of the right femur was cut to prepare unilateral bone defect models. Tricalcium phosphate bone graft was used in the experimental group, and no bone graft in the control group. During the fol owing 4 weeks, X-ray examination was done once a week to observe the bone healing, and then the animals were executed for col ecting samples in the graft area. The concentrations of bone morphogenetic protein-2 and vascular endothelial growth factor in samples which were taken from the bone graft area were determined by using ELISA assay. ＜br> RESULTS AND CONCLUSION:X-ray films showed that 2 weeks later, bone fracture healed mostly in the experimental group except a smal part of cortical bone;3 weeks later, bone fracture was basical y healed, and only a smal amount of tricalcium phosphate remained;4 weeks later, bone fracture was completely healed, and the cal us grew obviously, and the tricalcium phosphate was nearly absorbed. In the control group, the fracture line was stil visible at 1-2 weeks, but it became vague at 3 weeks;then, the fracture was healed at 4 weeks except some of the cortical bone. The levels of bone morphogenetic protein-2 and vascular endothelial growth factor were significantly higher in the experimental group than in the control group at different time points (P＜0.05). These results suggest that tricalcium phosphate bone graft can up-regulate the expression of bone morphogenetic protein-2 and vascular endothelial growth factor and accelerate bone healing.

Objective To investigate the clinical and genetic characteristics in two patients with androgen insensitivity syndrome. Methods Clinical features and laboratory data were collected from the patients and their families. All exons of the androgen receptor gene were amplified by PCR and PCR products were sequenced. Results Patient 1 presented with unambiguous female external genitalia, unilateral gynecomastia and primary amenorrhea. He did not have axillary hairs or pubic hairs. Patient 2 presented with undervirilization including scanty body hairs, gynecomastia and hypospadias. A missense mutation of

Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.

Objective To explore the antibacterial effect and mechanisms of platelets (PLT)on Staphylococcus epidermidis (SE).Methods Built infectious model of SE in vitro,whose final concentration was 105 CFU/ml in the reaction.Separately cocultured these models with PLT (the final concentration 400 × 109/L)-plasma (positive control)and BHI medium 12 hours later,and detected the antibacterial effect of PLT by making a liquid thinner and spreading counting method,and draw-ing antibacterial curve.Meanwhile,observed the bacterial structure by transmission electron microscope (TEM),and initially explored the antibacterial mechanism of PLT.Results The study showed the antibacterial effect of PLT on SE was very ob-vious,which appeared later than the plasma (M)group,but enduring.The images of TEM showed an electronlight region appeared in the centre of bacterium,contained condensed DNA molecules and leaded to slower fission.Conclusion PLT can damage the DNA structure of SE,and then affect the fission of SE,finally inhibit the proliferation of SE.

Objective To investigate whether miR?485?3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real?time PCR and Western blot were used to determine the expression of miR?485?3p and TLR1, respectively. The interaction between miR?485?3p and TLR1 was verified by target prediction software ( DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR?485?3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR?485?3p overexpression and TLR1 silencing on the activity of NF?κB. Western blot was used to study the effects of miR?485?3p overexpression and TLR1 silencing on NF?κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR?485?3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR?485?3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR?485?3p. miR?485?3p negatively regulated the expression of TLR1. The overexpression of miR?485?3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR?485?3p overexpression and TLR1 silencing reduced the activity of NF?κB and downregulated the expression of multiple NF?κB target genes. Conclusions miR?485?3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF?κB signaling pathway.

Objective To evaluate the relationship between CD4+ T lymphocyte count and results of enzyme-linked immunospot (ELISPOT) assay in human immunodeficiency virus (HIV)-Mycobacterium tuberculosis (M.tb) coinfected patients.Methods A total of 193 HIV-infected individuals in Yunnan Province and Shanghai were enrolled.T-SPOT.TB assay was employed to detect M.tb specific T lymphocyte in the peripheral blood mononuclear cells (PBMC).CD4+ T lymphocyte in PBMC from the enrolled subjects was detected by flow cytometry.Data were analyzed using t test.ResultsThe incidence of latent tuberculosis in HIV-infected individuals was 30.6％.The CD4+ T lymphocyte counts in HIV-infected individuals with active tuberculosis were 190×106/L,which were significantly lower than those in HIV-infected individuals with latent tuberculosis (484×106/L; t=6.665,P＜0.01).The HIV-infected individuals were stratified according to CD4+ T lymphocyte counts of ＞500×106/L,200×106-500×106/L,and ＜200×106/L and the constituent ratios of active tuberculosis/latent tuberculosis were 1∶16.2,1∶1.3 and 5.6∶1,respectively.Among 79 subjects with positive T-SPOT.TB results,20 were coinfected with active tuberculosis,in which 14 had CD4+ T lymphocyte counts of ＜200 ×106/L,5 had 200×105-500×106/L and 1 had ＞500×106/L.Fifty-two in 59 HIV/latent tuberculosis patients individuals had CD4+ T lymphocyte counts of ＞200×106/L.ConclusionsThe prevalence of latent tuberculosis in HIV-infected individuals is high in China.Cellular immunity in HIV-infected individuals with active tuberculosis is severely impaired.With the decrease of CD4 ′ T lymphocyte counts,patients with latent tuberculosis are prone to develop active tuberculosis in HIV-infected individuals.The negative predictive value of T-SPOT.TB is significantly diminished in patient with low CD4+ T lymphocyte counts,especially less than 200×106/L.

Objective To elucidate the effects of SP600125 at different concentrations on the proliferation and osteo-differentiation of human adipose-derived stem cells (hASCs).Methods The hASCs harvested were cocuhured with SP600125 at concentrations of 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmol/L in growth medium (OM group) and in osteogenesis medium (OM group),respectively.The DNA quantitative assay was carried out to evaluate proliferation of the hASCs;flow cytometry was used to determine the effect of SP600125 on the cell cycles of hASCs;Alkaline phosphatase level (ALP) and calcium deposition tests were conducted to observe the effects of SP600125 at different concentrations on osteogenic differentiation of the hASCs.Results The proliferation of hASCs was inhibited by 42.1％ when the cells were cocultured with SP600125 at the concentration of 10 μmol/L;the suppression decreased with decreased concentration of SP600125.The hASCs of phase G0/G1 in GM cocultured with SP600125 at the concentration of 10 μmol/L were more than those in GM cocultured with dimethylsulfoxide at the same concentration.ALP test revealed that after 10 days of culture in vitro the staining was more and more weakened and scattered and the ALP activity was more and more decreased with the increased concentration of SP600125.The extracellular calcium deposition of hASCs after 14 days of culture in vitro showed that the size and number of calcium nodules decreased with the increased concentration of SP600125.Conclusion SP600125 can suppress the proliferation and osteogenic differentiation of hASCs in vitro.

Dysglycemia is independently associated with the mortality of critically ill patients. Therefore, the management of blood glucose plays an important role in comprehensive therapy. It is suggested that the same target value of blood glucose (7.8-10.0 mmol/L) should not be set for all critically ill patients. Instead, it should be individually set based on the causes of the patient's admission and the status of blood glucose before admission. For this reason, there is an urgent need for a convenient protocol and method to regulate the dosage of insulin. The first hospital of Jiaxing, collaborating with information engineers, developed a modified eProtocol-insulin for domestic population with mathematical modeling and developed an Application Software (APP), which is convenient for clinical use. This is the first eProtocol-insulin and smart device APP for critically ill patients in China.

Objective:The study aimed to analyze the risk variables influencing the prognosis of patients with sepsis-associated delirium (SAD) in the Intensive Care Unit (ICU) and build a prediction nomogram.Methods:This was a retrospective cohort study that includes patients with SAD in the Medical Information Mart for Intensive CareⅢ database (MIMIC-Ⅲ) database as training cohort, and patients who were hospitalized in the First Hospital of Jiaxing from January 2021 to September 2022 as validation cohort. Inclusion criteria: (1) age≥18 years old; (2) being admitted to the ICU for the first time; (3) the length of ICU stay≥24 h; (4) consistent with the diagnosis of sepsis; (5) the diagnosis of delirium was identified by CAM-ICU questionnaire. The general information, vital signs, past medical history and laboratory examination results of the patients were collected, and the outcome was 28-day mortality. Multiple logistic regression was used to identify independent influencing factors and the nomogram was constructed. The validity of the prediction model was determined using multiple indicators, including calibration curve, the area under the receiver operating characteristic curve (AUC), decision curve analysis (DCA), and Hosmer-Lemeshow test.Results:A total of 250 patients were included in the training cohort and 154 patients were in the validation cohort. The multiple logistic regression demonstrated that age ( OR=1.057, 95% CI: 1.030-1.084, P<0.001), respiratory frequency ( OR=1.117, 95% CI: 1.037-1.202, P=0.003), lactic acid ( OR=1.137, 95% CI:1.011-1.279, P=0,032), hemoglobin ( OR=0.983, 95% CI: 0.970-0.997, P=0.020), SOFA score ( OR=1.184, 95% CI: 1.070-1.309, P=0.001) were independent risk factors associated with the 28-day mortality of patients with SAD. The AUC of the nomogram created by the five factors above was 0.773 (95% CI: 0.705-0.841), and the Hosmer-Lemeshow test showed that the model was a good fit ( P=0.875). The DCA curve indicates that the model has potential net benefit. The AUC was 0.864 (95% CI: 0.799-0.928) in the validation cohort, and the Hosmer-Lemeshow test showed that the model was a good fit ( P=0.488). The DCA curve indicates that the model of the validation cohort had potential net benefit. Conclusion:The prediction model based on age, respiratory frequency, lactate, hemoglobin, and SOFA scores shows valuable capability of predicting the prognosis of patients with SAD, which could help clinicians identify risk factors at first time and make earlier intervention.

Objective:To explore the effect and mechanism of intracellular Zn2+ concentration ([Zn2+ ]i) in hypoxia‐induced regulation of metalloproteinases (MMPs) and extracellular matrix (ECM) expression in nucleus pulposus (NP) cells .Methods:NP cells from SD rats received plate culture at first and then three‐dimensional culture with sodium alginate gel .[Zn2+ ]i was assayed by FluoZin‐3 AM staining .Proteoglycan was assayed by Alcian blue staining .Glycosaminoglycan was detected by 1 ,9‐dimethylmethylene blue (DMMB) assay .And real‐time PCR were used to assay the mRNA expression of α1 type II collagen (COL2A1) ,matrix metalloproteinase 13 (MMP‐13) and a disintegrin and metalloproteinase with a thrombospondin motif 5 (ADAMTS‐5) .The expression of ZRT ,IRT‐like protein 8(ZIP8) was assayed by immunohistochemistry and Western blotting .Results:Interleukin (IL)‐1βand ZnCl2 could significantly increase the [Zn2+ ]i of NP cells ,however ,the effect could be inhibited by hypoxia .Hypoxia did significantly attenuate the decrease of proteoglycan ,glycosaminoglycan ,and COL2A1 mRNA ,which was induced by IL‐1βand ZnCl2 treatment ,in sodium alginate three‐dimensional culture . However ,ZnCl2 inhibited the protective effect of hypoxia .Both an intracellular Zn2+chelator and hypoxia could inhibit the increase of MMP‐13 mRNA expression .IL‐1βand ZnCl2 treatment promoted the increase of ZIP8 expression in NP cells ,however ,hypoxia inhibited ZIP8 expression .Conclusions:Hypoxia may regulate the Zn2+ influx in NP cells . Zn2+ mediates the regulation effect of hypoxia on ECM and MMP‐13 .Perhaps the changes of [Zn2+ ]i are involved in the process of intervertebral disc degeneration .