Objective To establish a cisplatin-resistant 4T1 mouse model of triple negative breast cancer .Meth-ods A drug resistant mice model was established with cisplatin ( DDP ) induction and in-vivo/in-vitro tumorigenic ap-proach.Its resistance characteristics were identified by MTT assay .Changes of drug resistance gene ( MDR1, BCRP, MMP7, GST-π) and protein ( P-gp, BCRP, MMP7) expression, and phosphorate-Akt and total-Akt protein expression were evaluated by real-time PCR, immunohistochemistry and western blot method , respectively.Small animal live imaging technology was applied to detect tumor growth .Results Resistance fold (RF) of cisplatin-resistant 4T1 mouse model was 12.84.The expression of MDR1, BCRP, MMP 7, GST-πmRNA and P-gp, BCRP, MMP 7 proteins in the resistant mice were higher than that in the non-resistant mice .The result of western blot showed that a statistically higher expression of p-Akt in resistant mice than that in non-resistant mice at protein levels (P<0.01).No significant difference of tumor growth rate was observed between non-resistant and resistant mice ( P>0.05 ) .Given same dose of DDP , resistant mice showed lower sensitivity than non-resistant mice significantly (P<0.01).Conclusions We have successfully established a cis-platin-resistant triple negative breast cancer model in mice , which provides a new platform for further study on chemoresis-tant reversal strategy and individualized clinical treatment of this disease .

Objective To prepare recombinant human adenovirus type 3 expressing Norovirus cap-sid protein gene(Noro-orf2). Methods The cDNA for Noro-orf2 was amplifed by RT-PCR from stool of in-fantile gastroenteritis and cloned into the adenovirus shuttle vector pBSE3CMV-egfp. The vector pBSE3CMV-Nor was linearized with EeoR Ⅴ and Not Ⅰ, and transformed into E. coil BJ5183 with lined edenovirus ge-nomic DNA pLasmid pBRAdv3 by Rsr Ⅱ. The identification of recombinant adenovirus plasmid pBRAdv3E3dNor was performed by PCR, enzyme digestion and DNA sequencing. Then pBRAdv3E3dNor was digested with AsiS Ⅰ and transfeeted into Hep-2 cells with LipofectAMINETM 2000 to package recombi-nant adenovirus particles. Results Noro-orf2 was successfully inserted into the shuttle vector. The recombi-nant adenoviral plasmid pBRAdv3E3dNor was generated by homologous recombination in E. coil BJ5183 and confirmed by PCR and enzyme digestion. The recombinant adenovirus was successfully packaged and puri-fied. Norovirus eapsid protein gene expression was confirmed in Hep-2 cells by immunecytochemistry assay. Conclusion The recombinant type 3 adenovirus expressing Norovirus eapsid protein gene was successfully constructed. This study laid a foundation for developing vaccine against Norovirus.

To determine the bufalin concentration in rats' plasma by establishing a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method, and to evaluate and compare the pharmacokinetic characteristics of bufalin-loaded bovine serum albumin nanoparticles (bufalin-BSA-NP) and bufalin.

Objective To characterize the spinal motion of 6 degrees of freedom (6DOF) under the fulcrum bending in Lenke type 1 adolescent idiopathic scoliosis (AIS) patients using 3D image volume fusion technique.Methods Ten volunteers with Lenke type 1 AIS were examined by the Gemstone Spectral Imaging to obtain their T1-S1 vertebral morphological data in the supine position and fulcrum bending position.With the computer software,images of 2 different positions were fused with the 3D image to obtain in vivo kinematics data of each vertebral body in the target area.Results In the apical vertebrae (AV) region,the main range of motion in the fulcrum bending position ranged from-2.57°± 0.95° to-1.37°± 0.65°,showing significant differences between segments (P ＜ 0.05);along the vertical axis the coupling rotation ranged from-2.71° ± 0.48° to-1.27° ± 0.49°;along the coronal axis,the coupling flexion and extension ranged from 0.19° ± 0.85° to 1.51° ± 0.42°.In the upper end vertebrae (UEV) area,the main ROM ranged from-2.20° ± 1.13° to-1.91° ± 0.76°,showing no significant differences between the 3 segments (P ＞ 0.05);along the vertical axis,coupling rotation ranged from 0.18°±0.47° to 1.35°±0.52°;along the coronal axis,coupling flexion and extension ranged from 1.62° ± 0.41° to 2.07° ± 0.37°.In the lower end vertebrae (LEV) region,the main movement ranged from-0.79°± 0.63° to 0.73°± 1.09°,showing significant differences between the 3 segments (P ＜ 0.05);along the vertical axis,the coupling rotation ranged from-1.62°± 1.05° to-0.60°± 0.60°;along the coronal axis,the coupling flexion extension ranged from -1.41° ± 0.87° to-0.75° ± 0.66°.Conclusionts In the fulcrum bending position,Lenke type 1 AIS shows a unique motion and coupling mechanism in the spine.The coupling rotational movement in the UEV region is at the same side as the lateral flexion movement.The coupling rotational motion in the AV and LEV regions is opposite the lateral flexion movement.

Objective: To investigate the mechanism underlying the effects exerted by the Qizhu prescription (QZP) in breast cancer (BC), and the respective targets. Methods: Expression data from the ArrayExpress and The Cancer Genome Atlas (TCGA) were used to identify differentially expressed genes (DEGs) in BC. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the DEGs to identify genes involved in protein–protein interactions. Molecular docking was used to explore the dynamic relationship between active molecules and targets. Cell function experiments and animal studies were conducted to evaluate the effects of hub genes and active QZP compounds on BC cell behavior. Results: Among the 25 evaluated BC-related targets of QZP, matrix metalloproteinase-1 (MMP1) and epidermal growth factor receptor (EGFR) exhibited the highest degrees of dysregulation. GO and KEGG enrichment analyses revealed that the anti-BC targets of QZP primarily affected drug responses and pathways in cancer cells. Molecular docking analysis suggested potential interactions between EGFR and quercetin/luteolin, as well as between MMP1 and luteolin/kaempferol/quercetin. Quercetin significantly reduced BC cell proliferation, migration, invasion, and tumor development in vivo. Treatment of BC cells with quercetin decreased the expression or activation of several associated proteins. Conclusion The findings of our study provide new insights into the therapeutic potential of traditional Chinese medicine against BC, with particular reference to QZP.

Objective To investigate the efficacy and accuracy of 3D printing individualized navigation templates used in shoulder hemi-arthroplasty (HA).Methods We collected 9 adult cadaveric specimens of 18 shoulders with no obvious deformities or defects.After CT scanning was conducted on bilateral shoulders,the raw CT data in DICOM format were imported into software Mimics 10.01 and Imageware 12.0 to reconstruct 3D shoulder models.The retroversion angles and heights of bilateral humeral heads were measured to design individualized navigational templates using software Geomagic Studio 12.0.Rapid prototyping (RP) technique was used to manufacture the individualized navigation templates which were then used to assist shoulder HA in cadaveric specimens.CT scanning was performed postoperatively to measure the retroversion angle and height of the prosthesis implanted for comparison with the preoperative measurements.Results Before shoulder replacement in the 9 cadaveric specimens,respectively,the heights of left and right humeral heads averaged 300.0 ± 11.6 mm and 300.0 ± 11.4 mm,and the retroversion angles 15.6°±4.9° and 15.9°± 4.9°.After the replacement,the heights of left and right humeral heads averaged 300.0 ± 11.3 mm and 300.0 ± 11.7 mm,respectively,and the retroversion angles 15.7° ± 4.9° and 15.8° ± 5.2°,respectively.There were no significant differences between the left and right sides in the pre-or post-operative heights or retroversion angles of the humeral head (P ＞ 0.05).There were no significant differences either between preand post-operation in the heights or retroversion angles of the left and right humeral heads (P ＞ 0.05).The mean errors for the preoperative and postoperative heights and retroversion angles of the humeral heads were 1.44 ± 0.96 mm and 0.25° ± 0.15°,respectively.Conclusion Individualized navigation templates 3D printed can be used to assist effective and accurate shoulder HA because they are designed and manufactured according to accurate measurements of retroversion angles and heights of the humeral heads which are acquired by computer preoperatively.

The cgt gene was isolated from Paenibacillus macerans by PCR amplification and was inserted into vectors of pPIC9K and pMAS. The recombinant vectors were transformed to Pichia pastoris KM71 and Bacillus subtilis WB600, respectively. The results showed that alpha-CGTase activity in the culture media of recombinant P pastoris was only 0.2 U/mL, while it was 1.9 U/mL in recombinant B. subtilis. In addition, we optimized the culture conditions of the recombinant B. subtilis strain. After cultivation at 37 degrees C for 24 h with shake flask, the CGTase forming activity in culture media reached to 4.5 U/mL (hydrolysis activity was 3200 IU/mL), which is 9.8-fold to that of the original strain P. macerans.
Amino Acid Sequence ; Bacillus subtilis ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glucosyltransferases ; biosynthesis ; genetics ; metabolism ; Molecular Sequence Data ; Paenibacillus ; enzymology ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To investigate the clinical efficacy of oblique lateral interbody fusion (OLIF) combined with lateral plate (LP) fixation and posterior lumbar interbody fusion (PLIF) combined with pedicle screw fixation in the treatment of adjacent segment disease (ASDis).Methods:Data of 21 ASDis patients treated with OLIF-LP from August 2016 to October 2019 were selected, including 9 males and 12 females; age was 59.3±7.0 years (range, 46-71 years). Target segments: L 2, 3 1 cases (4.8%), L 3, 4 16 cases (76.2%), L 4, 5 4 cases (19.1%). Twenty-one ASDis patients matched with age, sex and surgical segment and treated with PLIF were selected as the control group. The operation time, intraoperative bleeding, postoperative hospital stay, visual analogua scale (VAS), Oswestry disability index (ODI), disc height (DH), intervertebral foramen height (IFH) and lumbar lordosis (LL) were compared between the groups were tested by t-test. VAS score, ODI, DH, IFH and LL were compared within the group by ANOVA, and Bonferroni's test was used for pairwise comparison. Results:All of 42 patients were followed up for 23.7 ±7.4 months (range, 12-36 months). The operation time (97.6 ± 18.0 min) and interpretative bleeding (38.5±62.7 ml) in OLIF-LP group were significantly lower than those in PLIF group (operation time 154.6±42.4 min) and interpretative bleeding (288.6±55.3 ml). There were significant differences between two groups ( t=5.66, P<0.001; t=8.23, P<0.001); the postoperative hospital stay 4.4±1.3 d in OLIF-LP group was longer than that in PLIF group 5.1±1.2 d, but there was no significant difference ( t=1.93, P=0.061); VAS score in OLIF-LP group at 1 month and 3 months after operation (1.6 ± 0.9 points, 1.4 ± 0.8 points), and the ODI index (29.4%±4.7%) after one month operation was improved better than that of PLIF group ( t=2.48, P=0.017; t=2.35, P=0.024; t=2.28, P=0.029), but there was no significant difference between the 12 months after operation of two groups ( t=0.99, P=0.329; t=0.86, P=0.395). The immediately after operation, 3 months after operation and 12 months after operation of DH, IFH and LL in the two groups were significantly improved compared with those before operation ( P<0.05). The immediately after operation, 3 months after operation and 12 months after operation of DH and IFH in the OLIF-LP group were better than those in the PLIF group ( P<0.05), while LL had no significant difference ( P>0.05). There were 2 cases (9.52%) in each group with cage sinking, but no clinical symptoms occurred. In the OLIF-LP group, there was no injury of blood vessels, nerves or abdominal organs during operation, and 2 patients had transient lower limb pain after operation; In the PLIF group, 2 cases (9.52%) of dural rupture were repaired during operation, and no cerebrospinal fluid leakage occurred after operation; Postoperative lower limb pain was aggravated in 3 cases, and improved after dehydration, anti-inflammatory and analgesic treatment; 2 cases of incision exudation healed after symptomatic treatment. Conclusion:OLIF combined with LP fixation has the same clinical effect as PLIF in the treatment of lumbar ASDis, but OLIF combined with LP fixation has more advantages in surgical trauma, postoperative recovery and related complications.

Objective:To investigate the characteristics of enterovirus (EV) VP1 gene from cases with hand, foot and mouth disease (HFMD) in Songjiang district of Shanghai in 2019.Methods:Samples from suspected HFMD cases were detected using real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR). Human rhabdomyosarcoma (RD) cells were used for EV culture. VP1 genes of the isolated EV A species were sequenced. The sequences of nucleotide and amino acid of EV A species were used in phylogenetic and homology analysis by MEGA X software.Results:Totally 207 HFMD specimens were detected in 2019, of which 188 specimens were screened positive for EV. The positive rates of Coxsackievirus (CV)-A6, CV-A16, CV-A10 and CV-A4 were 47.34% (89/188), 41.49% (78/188), 3.72% (7/188) and 2.66% (5/188), respectively. Other EV species were 4.79% (9/188) positive and EV-A71 was not detected. During the summer epidemic peak of HFMD (May to July), B1 gene subtype of CV-A16 was detected more frequently, of which B1a and B1b evolutionary branches were prevalent together. The CV-A6 virus of D3a branch dominated in secondary peak of autumn and winter (September to December). CV-A10 and CV-A4 were sporadic and both respective strains belonged to subtypes of C2 gene. Compared with the prototype strains, the nucleotide (amino acid) sequence homologies of CV-A6, CV-A16, CV-A10 and CV-A4 VP1 genes were 79.22%-81.78% (95.97%~-97.19%), 62.70%-65.54% (90.10%-91.30%), 81.76%-82.65% (91.63% -92.03%) and 81.09%-81.79% (97.27%-97.67%), respectively.Conclusions:The diversity and complexity of HFMD pathogen epidemic increase the difficulty of HFMD prevention and control. The expansion of EV surveillance programs and the studies on the molecular epidemiology of EV are helpful for the prevention and control of HFMD.

Objective:To explore the feasibility and clinical efficacy of prone transpsoas lateral interbody fusion (PTP LIF) combined with posterior pedicle screw fixation for the treatment of lumbar degenerative diseases in the prone position.Methods:A total of 23 patients who underwent LLIF in the prone position at the First People's Hospital of Yunnan Province between March 2023 and October 2023 were retrospectively analyzed. The cohort comprised 9 males and 14 females, with a mean age of 55.5±8.8 years (range, 41-70 years). The clinical diagnoses included intervertebral disc herniation with endplate inflammation (3 cases), lumbar spinal stenosis (13 cases), lumbar spondylolisthesis (5 cases), and lumbar instability (2 cases). The surgical segments involved L 3, 4 (15 cases), L 4, 5 (6 cases), and L 3-L 5 (2 cases), with 21 cases involving a single segment and 2 cases involving double segments. The disc height and lumbar lordosis Angle before and after surgery were compared. Lower back pain was evaluated using the visual analogue scale (VAS), while lumbar spine function was assessed via the Oswestry Disability Index (ODI). Clinical efficacy was evaluated according to the modified MacNab criteria at the last follow-up. Results:All surgeries were successfully completed. The operation time was 120.2±21.4 min (range, 90-175 min), intraoperative blood loss was 131.1±40.8 ml (range, 60-200 ml), and the hospital stay was 6.2±1.6 days (range, 4-10 days). Follow-up was obtained for all 23 cases, with the follow-up time being 9.6±2.2 months (range, 6-13 months). One case of endplate damage occurred during surgery, two cases of transient psoas muscle weakness occurred postoperatively, and one case of lower limb pain and numbness was reported; no cases of wound infection or delayed healing were observed. The postoperative disc height improved compared to preoperative (6.8±1.9 mm; F=66.618, P<0.001). There was no statistically significant difference between 3 months postoperative (11.1±1.2 mm) and immediately postoperative (12.2±1.2 mm; P>0.05), but there was a statistically significant difference between the last follow-up (10.7±1.1 mm) and immediately postoperative ( P<0.05). The postoperative lumbar lordosis angle improved compared to preoperative (35.3°±5.4°; F=19.465, P<0.001), with no statistically significant difference between 3 months postoperative (44.1°±5.4°) and immediately postoperative (47.8°±6.6°; P>0.05), but there was a statistically significant difference between the last follow-up (43.2°±5.3°) and immediately postoperative ( P<0.05). The postoperative VAS score improved compared to preoperative (6.3±1.1 points; F=79.931, P<0.001), and the last follow-up (1.1±1.1 points) showed further improvement compared to 3 months postoperative (1.7±1.4 points; P<0.05). The postoperative ODI improved compared to preoperative (69.9%±7.4%; F=592.392, P<0.001), with 3 months postoperative (23.1%±3.1%) showing improvement compared to 1 month postoperative (29.2%±3.1%), and the last follow-up (17.5%±3.6%) showing further improvement compared to 3 months postoperative ( P<0.05). At the last follow-up, the modified MacNab criteria were: excellent in 16 cases, good in 5, fair in 2, with an excellent and good rate of 91% (21/23); 7 cases of cage subsidence were observed, with no cases of internal fixation loosening. Conclusion:PTP LIF combined with pedicle screw fixation for the treatment of lumbar degenerative diseases is safe and effective, with satisfactory short-term postoperative outcomes.