Role of Perforating Artery Pedicled Neurotrophic Flap in the Treatment of Compound Tissue Defect of Tibia Using the Ilizarov Technique.

OBJECTIVE: To describe our experience with the combined use of pedicled neurotrophic flap and distraction osteogenesis in the management of complex lower extremity injuries with composite bone and soft tissue defects and assess the functional and cosmetic results of this method. METHODS: A pedicled flap with a marked perforator artery was applied for soft tissue coverage after radical debridement and temporary external fixation. In the second stage, the Ilizarov external fixator was used in place of the temporary external fixator for reconstruction of the segmental bone defect by distraction osteogenesis. Twenty-five patients (16 men and nine women; mean age, 39.2 years) were treated by using this combined technique between 2008 and 2016. All cases were graded initially as Gustilo-Anderson grade IIIB open fractures. The soft tissue defect after radical debridement ranged from 9 cm × 5 cm to 14 cm × 11 cm, and the average size of segmental defect was 5.2 (Range, 2.5-8.5) cm. Seventeen of these patients had a history of local infection. The bone structure and function were evaluated by two independent evaluators using Paley's criteria. RESULTS: Twenty-five patients were followed up for an average of 28.96 (Range, 15-48) months. The distally based sural neurovascular flap was applied in 13 patients, and the greater saphenous neurocutaneous perforator flap in 12 patients. The flap area ranged from 10 cm × 5 cm to 14 cm × 12 cm. Sufficient coverage of soft tissue defect was achieved in all cases. All flaps survived completely without complications. The bone defects were corrected by a mean lengthening of 6.94 (Range, 4.5-9.5) cm. The residual discrepancy was <1 cm in all cases, which was not clinically significant. The function was evaluated as excellent in 12 patients and good in 13 patients. Bone results were graded as excellent in 18 patients and good in seven patients. Complications during treatment included pain, pin tract infections, ankle midfoot joint stiffness, and docking site nonunion. No recurrence of infection was observed in infected patients. All cases achieved successful limb salvage and satisfactory function recovery without recurrence of infection. CONCLUSIONS: The combined technique of a perforator artery pedicled neurotrophic flap and distraction osteogenesis is an effective alternative approach in the salvage treatment of massively traumatized and chronically infected lower extremities.

Corrosion Resistance to Chloride of a Novel Stainless Steel: The Threshold Chloride Value and Effect of Surface State.

To evaluate the corrosion resistance of a novel stainless steel intended for use within reinforced concrete (RC) structures exposed to aggressive environments, the threshold chloride concentration of three stainless steels (316, 2205, novel 2205) and two carbon steels (HRB400, HRB500) exposed to pore solutions of fresh concrete was experimentally studied by means of electrochemical methods. The effect of steel surface state on the corrosion resistance was also experimentally investigated. The results showed that the novel stainless steel has a much higher corrosion resistance than those of the carbon steels and stainless steels when subjected to chloride environments. The presence of surface damage leads to significant decrease of corrosion resistance for carbon steel, however the corrosion can be certainly inhibited with the accumulation of rust on the steel surface. Although the oxide layer was worn, the novel 2205 stainless steel still has a great corrosion resistance.

Evaluation of a novel array-based toxoplasma, rubella, cytomegalovirus, and herpes simplex virus IgG enzyme linked immunosorbent assay and its comparison with virion/serion enzyme linked immunosorbent assays.

BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by κ-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with κ-coefficients showing near-perfect agreement for the HSV (κ=0.87), rubella (κ=0.92) and CMV (κ=0.93) and substantial agreement for the toxoplasma (κ=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.

Systematic expression analysis of genes related to multidrug-resistance in isogenic docetaxel- and adriamycin-resistant breast cancer cell lines.

Docetaxel (Doc) and adriamycin (Adr) are two of the most effective chemotherapeutic agents in the treatment of breast cancer. However, their efficacy is often limited by the emergence of multidrug resistance (MDR). The purpose of this study was to investigate MDR mechanisms through analyzing systematically the expression changes of genes related to MDR in the induction process of isogenic drug resistant MCF-7 cell lines. Isogenic resistant sublines selected at 100 and 200 nM Doc (MCF-7/100 nM Doc and MCF-7/200 nM Doc) or at 500 and 1,500 nM Adr (MCF-7/500 nM Adr and MCF-7/1,500 nM) were developed from human breast cancer parental cell line MCF-7, by exposing MCF-7 to gradually increasing concentrations of Doc or Adr in vitro. Cell growth curve, flow cytometry and MTT cytotoxicity assay were preformed to evaluate the MDR characteristics developed in the sublines. Some key genes on the pathways related to drug resistance (including drug-transporters: MDR1, MRP1 and BCRP; drug metabolizing-enzymes: CYP3A4 and glutathione S-transferases (GST) pi; target genes: topoisomerase II (TopoIIα) and Tubb3; apoptosis genes: Bcl-2 and Bax) were analyzed at RNA and protein expression levels by real time RT-qPCR and western blot, respectively. Compared to MCF-7/S (30.6 h), cell doubling time of MCF-7/Doc (41.6 h) and MCF-7/Adr (33.8 h) were both prolonged, and the cell proportion of resistant sublines in G1/G2 phase increased while that in S-phase decreased. MCF-7/100 nM Doc and MCF-7/200 nM Doc was 22- and 37-fold resistant to Doc, 18- and 32-fold to Adr, respectively. MCF-7/500 nM Adr and MCF-7/1,500 nM Adr was 61- and 274-fold resistant to Adr, three and 12-fold to Doc, respectively. Meantime, they also showed cross-resistance to the other anticancer drugs in different degrees. Compared to MCF-7/S, RT-qPCR and Western blot results revealed that the expression of MDR1, MRP1, BCRP, Tubb3 and Bcl-2 were elevated in both MCF-7/Doc and MCF-7/Adr, and TopoIIα, Bax were down-regulated in both the sublines, while CYP3A4, GST pi were increased only in MCF-7/Doc and MCF-7/Adr respectively. Furthermore, the changes above were dose-dependent. The established MCF-7/Doc or MCF-7/Adr has the typical MDR characteristics, which can be used as the models for resistance mechanism study. The acquired process of MCF-7/S resistance to Doc or Adr is gradual, and is complicated with the various pathways involved in. There are some common resistant mechanisms as well as own drug-specific changes between both the sublines.

Patterns of synonymous codon usage on human metapneumovirus and its influencing factors.

Human metapneumovirus (HMPV) is an important agent of acute respiratory tract infection in children, while its pathogenicity and molecular evolution are lacking. Herein, we firstly report the synonymous codon usage patterns of HMPV genome. The relative synonymous codon usage (RSCU) values, effective number of codon (ENC) values, nucleotide contents, and correlation analysis were performed among 17 available whole genome of HMPV, including different genotypes. All preferred codons in HMPV are ended with A/U nucleotide and exhibited a great association with its high proportion of these two nucleotides in their genomes. Mutation pressure rather than natural selection is the main influence factor that determines the bias of synonymous codon usage in HMPV. The complementary pattern of codon usage bias between HMPV and human cell was observed, and this phenomenon suggests that host cells might be also act as an important factor to affect the codon usage bias. Moreover, the codon usage biases in each HMPV genotypes are separated into different clades, which suggest that phylogenetic distance might involve in codon usage bias formation as well. These analyses of synonymous codon usage bias in HMPV provide more information for better understanding its evolution and pathogenicity.

Clonal spread of carbapenem non-susceptible Acinetobacter baumannii in an intensive care unit in a teaching hospital in China.

BACKGROUND: This study was aimed to investigate the genetic diversity and antibiotic resistance profile of the nosocomial infection agent Acinetobacter baumannii from a medical intensive care unit (ICU) in a teaching hospital in Suzhou, China. METHODS: The genetic relationship among A. baumannii isolates in an ICU was investigated using multilocus sequence typing (MLST). The antibiotic resistance pattern was determined by performing an antibiotic susceptible test, which included an agar dilution method and an E-test method. Resistant determinants, e.g., carbapenemase genes, metallo-ß-lactamases, and class 1 integron, were analyzed by specific PCR and DNA sequencing. RESULTS: In the present study, 33 non-duplicate isolates were identified as 5 existing sequence types (STs) (ST92, ST75, ST112, ST145, and ST345) and 1 new sequence type STn, which has a G-A mutation at nt268 on ropD40 of ST251. These results reveal limited diversity in carbapenem non-susceptible A. baumannii (CNSAb) isolates in our ICU, which are comprised of only 2 distinct STs, with ST92 and ST75 clustering into a clonal complex (CC) 92. Most CNSAb isolates (94.4%, 17/18) harbored the OXA-23 gene, while no carbapenem-susceptible A. baumannii (CSAb) isolates harbored it. In addition, 66.7% (22/33) isolates were positive for class 1 integrase, and gene cassette analysis showed there are 3 gene arrays among them, i.e., aacA4-catB8-aadA1 (77.3%, 17/22), aacA4 (22.7%, 5/22), and aacC1-orfX-orfX'-aadA1 (4.5%, 1/22). CONCLUSIONS: When all these data are combined, the antibiotic resistance and wide distribution of CNSAb isolates in our ICU are probably caused by expansion of the CC92 clone.

[Comparison of the clinical therapeutic effects between electroacupuncture and warming needle moxibustion for knee osteoarthritis of kidney deficiency and marrow insufficiency pattern/syndrome].

OBJECTIVE: To explore the actions of electroacupuncture (EA) and the warming needle moxibustion on knee osteoarthritis (KOA) of kidney deficiency and marrow insufficiency pattern/syndrome and compare the clinical effects between these two therapies. METHODS: Seventy-four cases of KOA were randomly divided into an electroacupuncture (EA) group and a warming needle moxibustion (WNM) group, 37 cases in each one. The acupoints were Dubi (ST 35), Neixiyan (EX-LE 4), Xuehai (SP 10), Zusanli (ST 36), Yanglingquan (GB 34), etc. In EA group, electric stimulation was given, 5 Hz, continuous wave. In the WNM group, warm needling technique was applied, 2 moxa cones on each acupoint in each time, three treatments a week. Totally, 4 weeks of treatment were required. The indicaices such as WOMAC score, illness severity index and systematic efficacy were adopted to evaluate the efficacy before treatment, 1 session and 2 sessions after treatment separately. RESULTS: The treatment in either group achieved the effectiveness. The cured and markedly effective rate was 64.7% (22/34) in EA group and was 40.0% (14/35) in WNM group, presenting statistically significant difference in comparison (P < 0.05). But the total effective rate did not indicate significance (P > 0.05). In EA group, the releasing effect of joint pain was obvious (P < 0.01). In the WNM group, the treatment was more advantageous at relieving joint stiffness (P < 0.01). There was no difference in the mean curative time between two groups (P > 0.05). CONCLUSION: Electroacupuncture and the warming needle moxibustion have their own advantages in the treatment of KOA of kidney deficiency and marrow insufficiency pattern/syndrome. Electroacupuncture is advantageous at analgesia and the warming needle moxibustion is at relieving joint stiffness. The total efficacy of electroacupuncture is superior to that of the warming needle moxibustion.

Use of Luminex xMAP bead-based suspension array for detecting microRNA in NSCLC tissues and its clinical application.

BACKGROUND: We measured the expression of microRNA (miRNA) in non-small cell lung cancer (NSCLC) tissues using the Luminex xMAP bead-based suspension array. We discuss the feasibility of employing this method to detect miRNA in NSCLC and explore its value as a high-throughput miRNA array. METHODS: We performed the methodological analysis of xMAP with oligoribonucleic acid references. We detected the expression of miR-21, miR, miR-31, miR-222, miR-145 and miR 40 NSCLC cancer tissues and adjacent normal tissues by xMAP bead-based suspension array. We selected miR-191 and miR-103 as the house-keeping genes (internal control). We also analyzed the relationship between xMAP and RT-PCR. RESULTS: The methodological analysis parameters of xMAP are quite good. The expression of miR-21, miR, miR-31 and miR-222 was higher in NSCLC tissues than in adjacent tissues, while the expression of miR-145 and miR-126 was lower in NSCLC tissues than in adjacent tissues. The expression of miR-145 and miR-126 decreased with disease progression. The intraassay and interassay coefficients of variation were lower in xMAP than in RT-PCR. xMAP proved cheaper and more flexible in detecting multiple miRNAs of one sample. CONCLUSIONS: The Luminex xMAP bead-based suspension array for detecting miRNA has many advantages, such as allowing a smaller sample size (only 2 µL), no sample amplification, fast detection, high throughput, and flexible combination of multiple detection targets. The high throughput testing technology shows a great advantage in saving time and labor. We found that the Luminex xMAP bead-based suspension array is a good and feasible method for detecting miRNA expression with high-throughput technology.

[Impact of acupoint heat-sensitive moxibustion on lung function and life quality of patients with chronic persistent bronchial asthma: a randomized controlled study].

OBJECTIVE: To observe the impact of heat-sensitive moxibustion on lung function in chronic persistent bronchial asthma and analyze its effect on the improvement of life quality as well as compare its efficacy with Seretide inhaler. METHODS: Fifty-seven cases were divided into a heat-sensitive moxibustion group (28 cases) and a Seretide group (29 cases) according to the random number table. In heat-sensitive moxibustion group, the therapy of heat-sensitive moxibustion was applied to the regions at the level of Feishu (BL 13) and Geshu (BL 17), or to the heat-sensitized points in the region 6 cun lateral from the 1st and 2nd intercostal spaces in the chest. The treatment was given continuously for 8 days, once per day, 12 treatments should be ensured in the later 22 days of the 1st month. In the later two months since then, 15 treatments should be ensured each month (< or = 1 treatment each day). In Seretide group, Seritide manufactured in GlaxoSmithKLine was used, one inhalation each time, twice per day. The cases in both groups were received treatment for 3 months. The changes in lung function such as forced expiratory volume one second (FEV1), TCM symptoms score, Asthma Control Test (ACT) and St George's Respiratory Questionnaire (SGRQ) were assessed and compared between two groups and within group separately. RESULTS: Lung function as FEV1 was improved in either group (P < 0.05, P < 0.01). The terminal improvement effects on symptoms of Chinese medicine, life quality and others were same between two groups (all P > 0.05). But the improvements in general situation, chills, fever and sweating in heat-sensitive moxibustion group were superior to those in Seretide group (both P < 0.05). Concerning to the improvement in respiratory symptoms, the effect of Seretide was better than that of heat-sensitive moxibustion (P < 0.05). CONCLUSION: Heat-sensitive moxibustion improves lung function, relieves clinical symptoms and benefits life quality for patients with chronic persistent bronchial asthma. It's efficacy on the disease is equal to the internationally-recognized effect of Seretide.

[Comparative research on effect of warm needling therapy on pulmonary function and life quality of patients with COPD in the stable phase].

OBJECTIVE: To observe the effects on chronic obstructive pulmonary disease (COPD) in the stable phase treated with warm needling therapy and Seretide. To make the comparison of improvements between two therapies in terms of the clinical symptoms and life quality. METHODS: Sixty cases were randomly divided into a warm needling group and a Seretide group, 30 cases in each one. In warm needling group, acupuncture with filiform needle was applied mainly to Dingchuan (EX-B 1), Feishu (BL 13) and Zusanli (ST 36). Warm needling therapy was supplemented on back-shu point and Zusanli (ST 36), once every other day, thrice each week. In Seretide group, Seretide inhalant was applied, one inhalation, 2 times each day. After 8-week treatment, the changes in pulmonary function such as forced expiratory volume in the first second (FEV1), clinical symptoms, and the score of St George's Respiratory Questionnaire (SGRQ) were observed in two groups and the comparison of them was conducted between groups and within each group. RESULTS: The pulmonary function indices such as FEV1 were all improved effectively in two groups (all P < 0.05). After 4-week treatment, the improvement in respiratory symptoms was superior in Seretide group as compared with warm needling group (P < 0.05). In terms of the improvement in general symptoms such as chills and fever, the result in warm needling group was better than that in Seretide group (P < 0.05). The efficacies were similar between two groups in 8-week treatment. Concerning to the improvement of life quality, the result in warm needling group was slightly better than that in Seretide group (the improvement value 12.8413.43 vs 10.21 +/- 3.89, P < 0.05). CONCLUSION: Warm needling therapy improves the pulmonary function of the patients with COPD in the stable phase, releases clinical symptoms and improves life quality. Its effects are similar to those of Seretide.

Effect of electro-acupuncture on tumor necrosis factor-α and vascular endothelial growth factor in peripheral blood and joint synovia of patients with rheumatoid arthritis.

OBJECTIVE: To observe the effect of electro-acupuncture (EA) on tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) in peripheral blood and joint synovia in patients with rheumatoid arthritis (RA) to verify the clinical efficacy of EA. METHODS: Adopting randomized controlled principle, the 63 RA patients enrolled were assigned to two groups, the 32 patients in the EA group were treated with EA, and the 31 patients in the simple needling (SN) group treated with simple needling. According to the integral-local combining method, the acupoints were selected mainly from yang-meridian and local Ashi points (pain-point). EA or SN was applied every other day, 10 times as a course, and each patient received a total of 3 courses of treatment. RESULTS: Blood and synovial levels of TNF-α and VEGF were reduced significantly after treatment in both groups. The lowering (absolute value and difference value) of TNF-α as well as the absolute value of VEGF, either in blood or in synovia, were similar in the two groups (P>0.05); but the lowering of VEGF after treatment was more significant in the EA group than that in the SN group (P<0.05). CONCLUSION: EA could effectively lower the contents of TNF-α and VEGF in peripheral blood and joint synovia to improve the internal environment for genesis and development of RA, so as to enhance the clinical therapeutic effectiveness.

[Effects of electroacupuncture and simple acupuncture on changes of IL-1, IL-4, IL-6 and IL-10 in peripheral blood and joint fluid in patients with rheumatoid arthritis].

OBJECTIVE: To explore the mechanism of electroacupuncture on rheumatoid arthritis (RA). METHODS: In a randomized and controlled trial, sixty-three cases with RA were randomly divided into an electroacupuncture group (n = 32) and a simple acupuncture group (n = 31). Baihui (GV 20), Fengchi (GB 20), Quchi (LI 11), Waiguan (TE 5), Guanyuan (CV 4) and Zusanli (ST 36) were selected by coordination method combined whole and local acupoints. The electroacupuncture group was treated with electroacupuncture at the local acupoints near painful joints, continuous wave, retaining needle for 30 minutes, and then electroacupuncture at Back-shu acupoints, retaining needle for 15 minutes, and the simple acupuncture group was treated with the same acupoints selection and acupuncture manipulation without electroacupuncture apparatus. They were all treated once every other day for 20 days as one course. After 3 courses, changes of interleukins in peripheral blood and joint fluid of patients were observed. RESULTS: Both of electroacupuncture and simple acupuncture had significant effect on IL-1, IL-4, IL-6 and IL-10 in peripheral blood and joint fluid of patients with RA ( P < 0.01, P < 0.05). But after electroacupuncture, the absolute value and improvement value of decreasing IL-1 in peripheral blood and joint fluid were super than those of simple acupuncture (all P < 0.05), and of IL-4 in joint fluid was super than that after simple acupuncture (P < 0.05), and of IL-6 and the absolute value of decreasing IL-10 were almost the same after both treatment (all P > 0.05), and after electroacupuncture, the improvement value of IL-10 in peripheral blood and joint fluid were super than those after simple acupuncture (both P < 0.05). CONCLUSION: Electroacupuncture can effectively decrease the proinflammatory cytokine of IL-1 and IL-6 and increase the inhibition cytokine of IL-4 and IL-10 and improve the internal environment of occurrence and progression of RA.