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.

A novel fully automated system for quantification of Hepatitis B virus DNA using magnetic bead-based method combined with real-time PCR.

BACKGROUND: Hepatitis B virus (HBV) infection is a major global health problem and causes liver damage as cirrhosis of the liver or liver cancer. Development of an accurate, sensitive and reproducible detection method for detecting and monitoring HBV DNA is very necessary and urgent. OBJECTIVES: The aims were to evaluate the analytical performances of the fully automated Pre-NAT system comparing to domestic assay, and to explore the role of highly sensitive quantification of HBV DNA in the management of chronic HBV infection. STUDY DESIGN: Pseudo-viral particles at high HBV DNA concentration were serially diluted to assess linear range. Accuracy and lower limit of detection were assessed by determining a panel of HBV standard substance. HBV DNA positive clinical specimen and internal quality control were measured 20 times to evaluate precision and reproducibility. 20 non HBV-infected specimens were used for the specificity assay. 96 chronic hepatitis B samples were quantified for HBV DNA to evaluating the correlation between the new test and Da-an assay. HBV serological markers were detected using ELISA method. RESULTS: Pre-NAT quantitated HBV DNA levels covered a wide dynamic range (10 logs) with a close correlation between expected and observed values (r=0.999, P<0.05), satisfactory precision and higher specificity. The lower detection limit was 20IU/mL. Comparability assay showed Pre-NAT had a good agreement with but more sensitive than Da-an assay (t=0.149, P>0.05). HBV DNA level was partially correlated to but more reliable and sensitive than serological evidence in reflecting the viral level. CONCLUSION: This novel fully-automated real-time PCR assay exhibits good analytical and clinical performances for highly sensitive detection of HBV DNA. It is well suited for monitoring antiviral responses and making treatment strategies according to current clinical practice guidelines for the management of chronic HBV infection.

Characteristics of carcinogenic human papillomavirus infection in Suzhou: Epidemiology, vaccine evaluation, and associated diseases.

Human papillomavirus infection is a major health problem and caused substantial benign and malignancy diseases among female and male worldwide. We aim to investigate the epidemiology of high-risk human papillomavirus (HR-HPV) and related diseases in Suzhou population. As well as evaluating the potential benefit of a nine-valent HPV vaccine (regardless of HPV-6 and -11) in Suzhou. A total of 40,108 people aged 13-89 years were retrospectively examined by database retrieval from 2010 to 2015. Thirteen genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 66) of HR-HPV were detected using Tellgenplex™ xMAP™ HPV DNA Test assay. The overall prevalence of HR-HPV was 21.1%, the female and male account for 96.4% and 3.6%, respectively. The infection rate among male (25.6%, 367/1,432) was significantly higher than that among female (20.9%, 8,100/38,676), X2 = 17.341 (P < 0.001), with OR = 1.293, 95% CI (1.146-1.460). The five most frequent HR-HPV genotypes were HPV-16 (5.12%), -52 (5.07%), -58 (3.02%), -39 (2.00%), and -18 (1.74%). HR-HPV infection rate was peak in person aged <20 years, and second higher in person aged 51-60 years. Infection modes as HPV-16, -18, -31, -33, -45, -52, -58 alone or mixed accounted for 63.2%. The top three prevalent diseases in HR-HPV infected women were cervicitis, vaginitis, and cervical lesions, and in men were verruca, urethritis, and balanitis, respectively. This is the first study to demonstrate HPV infection status in Suzhou population. Both women and men had a large burden of HPV infection. The nine-valent HPV prophylactic vaccines may potentially prevent 63.2% HR-HPV infection in Suzhou. J. Med. Virol. 89:895-901, 2017. © 2016 Wiley Periodicals, Inc.

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.

[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.

[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.