[Kirschner wire retractor-assisted reduction and inverted insertion of elastic nail in the treatment of children's irreducible subradial 1/3 fractures].

OBJECTIVE: To explore the clinical effect of Kirschner wire retractor-assisted reduction and inverted insertion of elastic nail in the treatment of children's irreducible subradial 1/3 fractures. METHODS: A total of 34 children with irreducible subradial 1/3 fractures treated by surgery from August 2016 to December 2020 were retrospective analyzed. Among them, 16 cases underwent Kirschner wire retractor-assisted closed reduction and percutaneous elastic intramedullary nailing with inverted insertion(observation group), 10 males and 6 females, aged from 4 to 10 years old with an average of(6.0±0.4)years;18 cases underwent open reduction and plate internal fixation (control group), 11 males and 7 females, the age from 3 to 10 years with an average of(7.0±0.5) years. The operation time, intraoperative blood loss, hospital stay, incision length, fracture healing time and complications of the two groups were observed and the wrist function was evaluated by Cooney wrist joint score. RESULTS: All patients were followed up for 3-12 years old with an average of (11.40±0.48) months in the observation group and 4-13 months with an average of (11.50±0.39) months in the control group. Bone healing was achieved in all patients, and there was no incision infection in both groups. The operation time, intraoperative blood loss, hospital stay and incision length in observation groups were lower than those of control group (P<0.05). There was no significant difference in the fracture healing time between two groups(P>0.05). There was no significant difference in postoperative healing and recovery of wrist function between groups(P>0.05). CONCLUSION: Compared with open reduction and plate internal fixation, Kirschner wire retractor-assisted reduction and percutaneous elastic intramedullary nail fixation for irreducible subradial radial 1/3 fractures has the advantages of less trauma, shorter operation time, less blood loss, and satisfactory short-term clinical results.

[Novel sternoclavicular hook-plate for treatment of proximal clavicle fracture with dislocation of sternoclavicular joint].

OBJECTIVE: To explore the efficacy of a novel sternoclavicular hook-plate for treatment of proximal clavicle fracture with dislocation of sternoclavicular joint. METHODS: Retrospective analysis of 13 cases of proximal clavicle fracture with dislocation of sternoclavicular joint treated with sternoclavicular hook-plate from June 2011 to January 2019 in our department. There were 9 males and 4 females, aged 26 to 78 years old, with an average age of (54.08±13.91) years old. All the patients had closed injuries without damage of blood vessels and nerves. The patient's operation time, intraoperative blood loss, hospital stay time, and postoperative complications were recorded. Fracture healing and reduction were evaluated according to X-ray and CT after operation. Constant-Murley score and Rockwood sternoclavicular joint score were used to evaluate limb function at 12 months after operation. RESULTS: All the patients were treated with sternoclavicular hook-plate. The operation time ranged from 50 to 76 min, with a mean of (54.08±13.91) min. The intraoperative blood loss ranged from 20 to 56 ml, with a mean of (46.08±11.15) ml. The hospital stay time ranged from 6 to 14 d, with a mean of (8.31±2.32) d. X-ray and CT examination on the second day after operation showed that all fractures and dislocations were anatomically reduced, and shoulder joint function exercise was performed early. All patients were followed up, and the duration ranged from 12 to 24 months, with a mean of (16.77±4.63) months. The healing time ranged from 9 to 13 d, with a mean of (11.00±1.75) d;and the bone healing time ranged from 3 to 4 months, with a mean of (3.65±0.46) months. There were no complications such as infection, internal fixation failure and nerve injury. At 12 months follow-up, the constant Murley score ranged from 78 to 100, with a mean of 87.83± 11.26; and Rockwood score ranged from 9 to 15, with a mean of 13.70±1.85. Among them, 11 cases were excellent, 1 case was good, and 1 case was general. CONCLUSION: The use of the novel sternoclavicular hook-plate for treatment of proximal clavicle fracture with dislocation of sternoclavicular joint is an effectively internal fixation with high safety, allowing early functional exercise for patients.

Psoas hematoma as a rare complication of posterior lumbar interbody fusion: a case report.

BACKGROUND: Psoas hematoma rarely occurs in patients with spondylolisthesis who undergo posterior lumbar interbody fusion (PLIF) surgery. CASE PRESENTATION: Here we reported a case of a 57-year-old male patient diagnosed with spondylolisthesis who underwent PLIF at the local hospital. Seven days post-surgery, abdominal pain occurred, and the pain in the right lower limb gradually increased. The computerized tomography (CT) indicated a formation of hematoma around the psoas muscle. Digital-subtraction angiography (DSA) suggested a vascular injury, a rupture of the right segmental artery of the lumbar vertebral level 4. The patient then received DSA vascular embolization, after which the lower lumbar segmental artery active bleeding was stopped. One month after discharge, the abdominal hematoma was gradually absorbed, and the pain in the waist, leg, and abdomen disappeared. CONCLUSION: Symptoms such as abdominal pain, abdominal distension, and exacerbation of lower limb pain, may suggest the occurrence of psoas hematoma after PLIF. DSA vascular embolization is suggested as the first treatment approach for this type of complication.

Case report: small cell transformation and metastasis to the breast in a patient with lung adenocarcinoma following maintenance treatment with epidermal growth factor receptor tyrosine kinase inhibitors.

BACKGROUND: Breast metastasis from lung cancer has been reported, but not from SCLC that is transformed from lung adenocarcinoma during maintenance treatment with epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). Transformation to small cell lung cancer(SCLC), although uncommonly seen, has been associated with resistance to EGFR-TKI therapy in lung adenocarcinomas. CASE PRESENTATION: We describe a case of a 49-year-old man with lung adenocarcinoma harboring L858R point mutation at the exon 21 of the epidermal growth factor receptor (EGFR). During the maintenance treatment with EGFR-TKI, the patient presented with a right breast mass, which was accompanied by elevated serum neuron specific enolase (NSE) level. The histological examination of biopsies from the breast mass and enlarging lung mass revealed SCLC that was less sensitive to standard SCLC treatment. The breast tumor was positive for thyroid transcription factor-1 (TTF-1), consistent with a lung primary cancer. CONCLUSION: This is the first case report of small cell transformation and metastatic to the breast in a patient with lung adenocarcinoma following EGFR-TKI treatment. Repeat biopsy is important for evaluation of evolving genetic and histologic changes and selection of appropriate treatment. and serum NSE measurement may be useful for detection of small cell transformation in cases with resistance to EGFR-TKI therapy.

Construction of Radial Defect Models in Rabbits to Determine the Critical Size Defects.

Many studies aimed at investigating bone repair have been conducted through animal models in recent years. However, limitations do exist in these models due to varying regeneration potential among different animal species. Even using the same animal, big differences exist in the size of critical size defects (CSD) involving the same region. This study aimed to investigate the standardization of radial bone defect models in rabbits and further establish more reliable CSD data. A total of 40 6-month-old New Zealand white rabbits of clean grade totaling 80 radial bones were prepared for bone defect models, according to the principle of randomization. Five different sizes (1.0, 1.2, 1.4, 1.7 and 2.0 cm) of complete periosteal defects were introduced under anesthesia. At 12 weeks postoperatively, with the gradual increase in defect size, the grades of bone growth were significantly decreased in all 5 groups. X-ray, CT scans and H&E staining of the 1.4, 1.7, and 2.0-cm groups showed lower grades of bone growth than that of the 1.0 and 1.2-cm groups respectively (P < 0.05). Using rabbit radial defect model involving 6-month-old healthy New Zealand white rabbits, this study indicates that in order to be critical sized, defects must be greater than 1.4 cm.

Efficacy and safety of ifosfamide-based chemotherapy for osteosarcoma: a meta-analysis.

BACKGROUND: The efficacy of ifosfamide-based chemotherapy in the treatment of osteosarcoma has been investigated; however, results are inconsistent. Therefore, we reviewed the relevant studies and conducted a meta-analysis to assess the efficacy of ifosfamide-based chemotherapy in patients with osteosarcoma. METHODS: A systematic literature search on PubMed, Embase, and Web of Science databases was performed. Eligible studies were clinical trials of patients with osteosarcoma who received ifosfamide-based chemotherapy. Hazard ratios (HRs) were pooled to compare event-free survival (EFS) and overall survival (OS). Risk ratios (RRs) were pooled to compare good histologic response rates and adverse event incidence. Meta-analysis was performed using a fixed-effects model or a random-effects model according to heterogeneity. RESULTS: A total of seven randomized controlled trials were included in this meta-analysis. Pooled results showed that ifosfamide-based chemotherapy significantly improved EFS (HR=0.72, 95% confidence interval [CI]: 0.63, 0.82; P=0.000) and OS (HR=0.83, 95% CI: 0.70, 0.99; P=0.034); furthermore, this form of chemotherapy increased good histologic response rate (RR=1.27, 95% CI: 1.10, 1.46; P=0.001). In addition, patients in the ifosfamide group exhibited a significantly higher incidence of fever (RR=2.23, 95% CI: 1.42, 3.50; P=0.000) and required more frequent platelet transfusion (RR=1.92, 95% CI: 1.23, 3.01; P=0.004). CONCLUSION: This meta-analysis confirmed that ifosfamide-based chemotherapy can significantly improve EFS and OS; this chemotherapy can also increase good histologic response rate in patients with osteosarcoma. However, evidence may be limited by potential biases and confounders. Thus, large-scale well-designed randomized controlled trials are needed to verify current findings.

Suppressive effects of quercetin-3-O-(6″-Feruloyl)-ß-D-galactopyranoside on adipogenesis in 3T3-L1 preadipocytes through down-regulation of PPARγ and C/EBPα expression.

Obesity is a chronic, costly disease, and flavonoids such as quercetin have been proven to play protective roles against it. This study investigated the suppressive effect of quercetin-3-O-(6″-feruloyl)-ß-D-galactopyranoside (QFG) on adipogenesis of 3T3-L1 preadipocytes. Quercetin-3-O-(6″-feruloyl)-ß-D-galactopyranoside and quercetin were both extracted from Psidium guajava (Myrtaceae, commonly known as guava) leaves and were evaluated for their suppressive effect on adipogenesis by means of oil red O staining and triglyceride assay. It was shown that QFG inhibited adipogenesis in a dose- and time-dependent manner, and it exerted a stronger effect than did quercetin at the same concentration. Quantitative real-time polymerase chain reaction and western blotting were conducted to further examine the differentiation expression of marker genes and transcriptional factors. Both mRNA and protein expression of the key adipogenic transcriptional factors, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT (cytidine-cytidine-adenosine-adenosine-thymidine)/enhancer-binding protein alpha (C/EBPα), were inhibited by QFG. Moreover, the mRNA expression patterns of key participants in the Wnt-ß-catenin pathway were not altered during the QFG-induced adipogenesis inhibition. These results suggest that QFG effectively suppresses adipogenesis and that it exerts its role mainly through the significant down-regulation of PPARγ and C/EBPα and, probably, via a Wnt-ß-catenin independent pathway.

Maohuoside A promotes osteogenesis of rat mesenchymal stem cells via BMP and MAPK signaling pathways.

Osteoporosis is becoming a more prevalent health problem with the aging of the population around the world. Epimedium koreanum Nakai is one of the most used herbs in East Asia for curing osteoporosis, with its major ingredient, icariin, mostly explored by researchers. In this article, maohuoside A (MHA), a single isolated compound from the herb, was identified to be more potent than icariin in promoting osteogenesis of rat bone marrow-derived mesenchymal stem cells (rMSCs) (increasing by 16.6, 33.3, and 15.8% on D3, D7, and D11, respectively). Alkaline phosphatase (ALP) assay and calcium content measurement were assigned to quantify the promoted osteogenesis and alizarin red S (ARS) staining was conducted to visualize it. Quantitative real-time PCR (Q-PCR) was assayed to evaluate the mRNA expression of marker genes in osteogenesis and master regulators in BMP pathway. Moreover, PD98059 (PD) and SB203580 (SB), inhibitor of ERK1/2 and p38 MAPK pathway, were administered to assess the involvement of MAPK pathway in the promotion process. In conclusion, MHA pronouncedly enhanced the osteogenesis of rMSC, plausibly via the BMP and MAPK signaling pathways.

Transient expression of interferon-inducible p204 in the early stage is required for adipogenesis in 3T3-L1 cells.

A member of the interferon-inducible p200 family of proteins, p204, has recently been reported to function in the development of many mesoderm-derived tissues, such as bone, muscle, and cartilage. However, no published study has yet investigated the role of p204 in adipogenesis. Our preliminary experiments showed that p204 can be found in 3T3-L1 preadipocytes, and its expression was up-regulated in a differentiation-dependent manner. As such, we hypothesized that p204 is associated with adipogenesis and focused on the influence of p204 on adipogenesis. In the present study, we investigated the transient elevated expression and cytoplasm-to-nucleus translocation of p204 in the early stage of adipogenesis. To determine the effect of p204 on adipogenesis, p204-siRNA and expression vector were produced for p204 suppression and overexpression, respectively. The knockdown of p204 resulted in a significantly depressed adipocyte differentiation, whereas p204 overexpression promoted adipocyte differentiation. The mRNA expression of adipogenic markers, such as peroxisome-proliferator-activated receptor (PPAR)gamma, CCAAT/enhancer-binding-protein (C/EBP)alpha, lipoprotein lipase, and adipsin, was decreased by p204 suppression and increased by p204 overexpression. A coimmunoprecipitation assay coupled with an indirect immunofluorescence assay also indicated that p204 interacted and colocalized with C/EBPdelta in the nucleus. Furthermore, the knockdown of p204 disrupted the interaction between p204 and C/EBPdelta and partially suppressed the PPARgamma transcriptional activity by dissociating C/EBPdelta with the PPARgamma promoter element. Collectively, our data indicate that the transient expression of p204 in the early stage is indispensable for adipocyte differentiation. Disruption of p204 expression patterns at this stage leads to irreversible damage in fat formation.

Estrogen receptor mediates the effects of pseudoprotodiocsin on adipogenesis in 3T3-L1 cells.

Estrogen receptors (ERs) play a pivotal role in adipogenesis; therefore, compounds targeting ERs may also affect fat formation. Recent studies have shown that the Dioscorea plant (commonly called yam) exhibits an antiobesity effect on rodents. However, the active compounds and underlying mechanisms responsible for this effect are not yet fully understood. We evaluated the effects of pseudoprotodiocsin (PPD), a steroid saponin from Dioscorea nipponica Makino (a type of Dioscorea), on adipogenesis and the mechanisms underlying this effect. Treatment with PPD at the onset of adipogenic differentiation resulted in significantly decreased adipogenesis in both in vitro and in vivo experimental systems. An increased amount of ERalpha mRNA, protein, and the accumulation of ERalpha in the nucleus were also observed. However, the expression pattern of ERbeta was not altered. Furthermore, the antiadipogenic effect of PPD was found to be ER dependent. It was also accompanied by the decreased expression of several genes involved in adipogenesis, including lipoprotein lipase (LPL), leptin, CCAAT/enhancer-binding-protein-alpha (C/EBPalpha), and peroxisome proliferator-activated receptor-gamma (PPARgamma), as well as the increased expression of some negative factors of adipogenesis, including preadipocyte factor 1 (Pre-1), GATA-binding protein 2 (GATA-2), GC-induced leucine-zipper protein (GILZ), and C/EBP homologous protein (CHOP-10). In addition to its estrogenic action, PPD also abolished the p38 mitogen-activated protein kinase (p38 MAPK) activation. Our results suggest that PPD inhibits adipogenesis in an ER-dependent manner and induces the expression of ERalpha. These findings may provide a lead toward a novel agent that can be used to treat obesity.

Isolation of a new glycoside from starfish, Asterias amurensis Lutken, and its stimulation activity on the proliferation of the UMR106 cell.

A new glycoside compound (1) was isolated from the starfish Asteria amurensis Lutken. The structure for compound 1 was identified as 1-O-{beta-D-quinovopyranosyl-(1-2)-beta-D-fucopyranosyl-(1-4)-[beta-D-fucopyranosyl(1-2)] beta-D-quinovopyranosyl}-butanol by extensive NMR experiments as well as chemical evidence. The effects of compound 1 on UMR106 cell proliferation were screened by MTT assay. The results indicate that compound 1 (0.01-100 microM) significantly promotes osteoblastic proliferation.

[Preparation and application of anti-serum against mouse visfatin].

AIM: To prepare polyclonal anti-serum against mouse visfatin and explore the expression of visfatin in different mouse tissue. METHODS: A fragment of gene coding the full length of visfatin was amplified and cloned into the prokaryotic expression vector pET28a. Then the recombined protein was expressed in E.coli(BL21) and purified by Ni-NTA HIS*BIND RESIN. After the indentification of Western blot, the recombined protein was used to immune New Zealand rabbit. With the purified anti-serum, we characterized the expression of visfatin in different mouse tissues. RESULTS: According to the results of SDS-PAGE and Western blot, the expression of recombined prokaryotic expression vector pET28a-visfatin could be induced by IPTG in E.coli. The titer of the anti-serum was above 1:25 600. CONCLUSION: The results for the identification of vifatin among mouse tissues show that visfatin exists in all the detected tissues, with the expression in spleen and adipose tissue higher than the others.

[Construction of eukaryotic expression vector of rat HAS-3 gene and its effects on chemotaxis].

AIM: To construct the eukaryotic expression vector of rat HAS-3 gene, express it in RSC96 cells and detect the enzyme activity of the recombinant protein. METHODS: Rat HAS-3 gene was cloned from CCI rat injury nerve cDNA using RT-PCR and inserted into pcDNA3.1D and pEGFP-N1. The recombinant plasmids were transfected to RSC96 cells. The expression level and enzyme activities of the recombinant proteins were monitored. RESULTS: Rat HAS-3 gene was cloned into pcDNA3.1D and pEGFP-N1 correctly. Recombinant proteins were detected in RSC96 cells and the synthesis of HA was up-regulated after transcfection. The condition medium of RSC96 cells overexpressing HAS-3 had some effects on chemotaxis of macrophages. CONCLUSION: The eukaryotic expression vectors of rat HAS-3 has been constructed successfully. The transfection of HAS-3 gene to RSC96 cells is effective in chemotaxis of macrophages which demonstrates HAS-3 may contribute to the development of inflammation in vivo.

Two new steroidal compounds from starfish Asterias amurensis Lutken.

Two new sulfated steroidal compounds (1 and 2), along with three known steroidal saponins (3, 4, and 5) were isolated from the starfish Asterias amurensis Lutken. The structures of new compounds were elucidated as 3beta-O-sulfated-cholest-5-ene-7alpha-ol (1) and (E) 25-O-beta-d-xylopyranosyl-26, 27-dinor-24(S)-methyl-22-ene-15alpha-O-sulfated-5alpha-cholest-3beta,6alpha-ol (2) by extensive NMR experiments and chemical evidence. Their effects on UMR106 cell proliferation were screened by MTT method. The results indicated that compounds 2 and 2a (0.01-100 microM) significantly promoted the osteoblastic proliferation. The initial structure-activity relationship analysis suggests that the sugar moiety is the necessary group for the activity.

Apoptosis induced by (di-isopropyloxyphoryl-Trp)2-Lys-OCH3 in K562 and HeLa cells.

According to the method used in our laboratory,our group synthesized (DIPP-Trp)2-Lys-OCH 3. It inhibited the proliferation of K562 and HeLa cells in a dose-and time-dependent manner with an IC 50 of 15.12 and 42.23 microM, respectively. (DIPP-Trp) 2-Lys-OCH3 induced a dose-dependent increase of the G2/M cell population in K562 cells, and S cell population in HeLa cells;the sub-G0 population increased dramatically in both cell lines as seen by PI staining experiments using a FACS Calibur Flow cytometer (BeckmanCoulter,USA). Phosphatidylserine could signi?cantly translocate to the surface of the membrane in (DIPP-Trp)2-Lys-OCH3-treated K562 and HeLa cells. The increase of an early apoptotic population was observed in a dose-dependent manner by both annexin-FITC and PI staining. It was concluded that (DIPP-Trp) 2-Lys-OCH3 not only induced cells to enter into apoptosis,but also affected the progress of the cell cycle. It may have arrested the K562 and HeLa cells in the G 2/M,S phases,respectively. The apoptotic pathway was pulsed at this point,resulting in the treated cells entering into programmed cell death.(DIPP- Trp)-Lys-OCH is a potential anticancer drug that intervenes in the signalling pathway.

[Interleukin-8 inhibits clonal expansion of 3T3-L1 preadipocyte during differentiation].

AIM: To investigate the effect of interleukin-8 (IL-8) on the differentiation and clonal expansion of 3T3-L1 preadipocyte during the differentiation period. METHODS: The morphological changes of 3T3-L1 cells during differentiation after the treatment of IL-8 was observed by Oil-Red O staining. Glycerol-3-phosphate dehydrogenase (GPDH) activity was measured by a spectrophotometric method. MTT method and 3H-TdR incorporation were applied to examine the changes of cell proliferation and DNA synthesis in clonal expansion of 3T3-L1 cells. Cell cycle analysis was taken by flow cytometry. RESULTS: IL-8 could inhibit the differentiation and GDPH activity in a dose dependent manner. IL-8 decreased the cell proliferation and DNA synthesis in clonal expansion after induction. Also, the proportion of cells in G1 phase was increased and that of cells in S and G2 phase was declined after the treatment of IL-8. CONCLUSION: IL-8 inhibits the differentiation of 3T3-L1 preadipocytes by decreasing the clonal expansion of the cells.

Molecular cloning and characterization of rat chitotriosidase.

Recent year some members of mammalian chitinases and chitinase-like proteins have been discovered, but rat counterpart of human and mouse chitotriosidase has not been identified. Moreover, the physiological functions of mammalian chitinases are not very clear. To facilitate the studies we cloned the cDNA encodes the rat chitotriosidase. The results revealed that it is differ from mouse and human chitotriosidase genes, it exist alternative splicing transcripts in several tissues we detected due to different transcriptional initiation sites and different exon usage, although all the open reading frame of these cDNAs predict a protein of 464 amino acids with a typical chitinase structure, including a signal peptide, a highly conserved catalytical domain and a chitin-binding structure. The predicted amino acid sequence is highly homologous to that of mouse and human chitotriosidase. Recombinant expression of the cloned cDNA demonstrated that the encoded protein is secreted extracellularly and has chitinolytic activity.