Combinatorial metabolic engineering of Bacillus subtilis for de novo production of polymyxin B.

Polymyxin is a lipopeptide antibiotic that is effective against multidrug-resistant Gram-negative bacteria. However, its clinical development is limited due to low titer and the presence of homologs. To address this, the polymyxin gene cluster was integrated into Bacillus subtilis, and sfp from Paenibacillus polymyxa was expressed heterologously, enabling recombinant B. subtilis to synthesize polymyxin B. Regulating NRPS domain inhibited formation of polymyxin B2 and B3. The production of polymyxin B increased to 329.7 mg/L by replacing the native promoters of pmxA, pmxB, and pmxE with PfusA, C2up, and PfusA, respectively. Further enhancement in this production, up to 616.1 mg/L, was achieved by improving the synthesis ability of 6-methyloctanoic acid compared to the original strain expressing polymyxin heterologously. Additionally, incorporating an anikasin-derived domain into the hybrid nonribosomal peptide synthase of polymyxin increased the B1 ratio in polymyxin B from 57.5% to 62.2%. Through optimization of peptone supply in the fermentation medium and fermentation in a 5.0-L bioreactor, the final polymyxin B titer reached 962.1 mg/L, with a yield of 19.24 mg/g maltodextrin and a productivity of 10.02 mg/(L·h). This study demonstrates a successful approach for enhancing polymyxin B production and increasing the B1 ratio through combinatorial metabolic engineering.

Validating the Role of Tibial Tubercle-Posterior Cruciate Ligament Distance and Tibial Tubercle-Trochlear Groove Distance Measured by Magnetic Resonance Imaging in Patients With Patellar Dislocation: A Diagnostic Study.

PURPOSE: To (1) compare these parameters between patients with patellar dislocation and healthy individuals on magnetic resonance imaging measurements, (2) validate the diagnostic capacity of the tibial tubercle-posterior cruciate ligament (TT-PCL) distance and tibial tubercle-trochlear groove (TT-TG) distance, (3) define the pathologic threshold values of these parameters for tibial tubercle osteotomy indication, and (4) compare these values with those of previous studies. METHODS: Seventy patients with patellar dislocation and 70 healthy volunteers were identified. The inter- and intraobserver reliability values were determined using Bland-Altman analysis and the intraclass correlation coefficient (ICC). The diagnostic capacity of the parameters was evaluated using receiver operating characteristic curves and the area under the receiver operating characteristic curve. The data of the control group were used to determine the pathologic threshold values of the measurements. Logistic regression analysis was performed with these pathologic threshold values. RESULTS: Significant differences in the TT-PCL distance (P = .01) and TT-TG distance (P < .001) were found between the study group (21.48 ± 3.18 and 12.91 ± 3.80, respectively) and the control group (20.07 ± 2.99 and 8.46 ± 3.16, respectively). Both the TT-PCL distance and TT-TG distance had excellent inter- and intraobserver agreement, with inter-ICCs >0.915 and intra-ICCs >0.932, respectively. However, the TT-TG distance had a higher area under the receiver operating characteristic curve than did the TT-PCL distance (0.820 vs 0.627). The pathologic threshold value of the TT-PCL distance was 24.76 mm. The pathologic threshold value of the TT-TG distance was 13.64 mm. Subjects with a TT-TG distance of >13.64 mm had a greater risk for patellar dislocation, with an odds ratio of 14.02 (95% confidence interval 4.00-49.08, P < .001). CONCLUSIONS: Both the TT-PCL distance and TT-TG distance can be measured reliably by magnetic resonance imaging; however, the TT-TG distance has a better diagnostic capacity than does the TT-PCL distance. LEVEL OF EVIDENCE: Retrospective cohort study (diagnosis); Level of evidence, I.

Control of the polymyxin analog ratio by domain swapping in the nonribosomal peptide synthetase of Paenibacillus polymyxa.

Polymyxins are used as the last-line therapy against multidrug-resistant bacteria. However, their further clinical development needs to solve problems related to the presence of heterogeneous analogs, but there is still no platform or methods that can regulate the biosynthesis of polymyxin analogs. In this study, we present an approach to swap domains in the polymyxin gene cluster to regulate the production of different analogs. Following adenylation domain swapping, the proportion of polymyxin B1 increased from 41.36 to 52.90%, while that of B1-1 decreased from 18.25 to 3.09%. The ratio of polymyxin B1 and B3 following starter condensation domain swapping changed from 41.36 and 16.99 to 55.03 and 6.39%, respectively. The two domain-swapping strains produced 62.96% of polymyxin B1, 6.70% of B3 and 3.32% of B1-1. This study also revealed the presence of overflow fluxes between acetoin, 2,3-butanediol and polymyxin. To our best knowledge, this is the first report of engineering the polymyxin synthetase gene cluster in situ to regulate the relative proportions of polymyxin analogs. This research paves a way for regulating lipopeptide analogs and will facilitate the development of novel lipopeptide derivatives.

Novel 2, 8-bit derivatives of quinolines attenuate Pseudomonas aeruginosa virulence and biofilm formation.

Signal molecules are stimulators of multiple quroum-sensing virulence and biofilm formation. Small molecule analogues have been suspected as a potent inhibitor in therapeutic strategy. Herein, we synthesized a series of small molecule compounds from the 2, 8-bit derivatives of quinoline by Suzuki coupling reaction. We found that these compounds have the biofilm inhibitory effect in normal condition instead of phosphate limitation state. Furthermore, lacZ reporter strain assay and rhamnolipids as well as pyocyanin experiments showed that these compounds did not affect las and pqs system but reduced the expression of rhl. All these results suggest that quinoline derivatives can be treated as potent inhibitors against biofilm and reduce virulence through the rhl system. This research will be useful in designing new quorum sensing inhibitors to attenuate the infection of bacteria.

The prevalence and morphological characteristics of the knee anterolateral ligament in a Chinese population.

The anterolateral ligament, a distinct structure connecting the lateral femoral epicondyle to the anterolateral proximal tibia, is gaining attention because of its possible function in ensuring internal rotational stability of the tibia. To study the prevalence and precise anatomical characteristics of the anterolateral ligament and its relationship to adjacent structures in a Chinese population, a total of 20 amputated knee specimens were collected. The anterolateral regions of the knees underwent detailed surgical dissection, followed by precise measurement of the anterolateral ligament and its adjacent structures. Histological analysis of the anterolateral ligament was performed using hematoxylin and eosin (H&E) staining. A thin soft tissue deep to the iliotibial band running obliquely across the lateral fibula ligament and connecting the lateral head of the gastrocnemius with the tibia, termed the 'gastrocnemius-tibial ligament' or superficial layer of the anterolateral ligament, was observed in 18 of the 20 specimens, corresponding to a prevalence of 90%. Furthermore, a well-defined anterolateral ligament deep to the gastrocnemius-tibial ligament and distinct from the lateral fibula ligament was found in all 20 knees (prevalence, 100%). The independent gastrocnemius-tibial ligament and anterolateral ligament had separate femoral originations at the lateral head of the gastrocnemius and the lateral femoral epicondyle, and the same osseous tibial insertion at the midpoint between Gerdy's tubercle and the most lateral aspect of the fibular head. H&E staining showed that both the anterolateral ligament and gastrocnemius-tibial ligament were ligaments consisting of collagenous bundles. In the Chinese Han population, the gastrocnemius-tibial ligament and anterolateral ligament may form a complex at the anterolateral aspect of the knee, which is likely involved in ensuring the internal rotational stability of the tibia.

Anatomic Anterolateral Ligament Reconstruction Improves Postoperative Clinical Outcomes Combined with Anatomic Anterior Cruciate Ligament Reconstruction.

A significant cohort of patients is plagued by postoperative rotational instability after the anatomic anterior cruciate ligament (ACL) reconstruction surgery. Anatomic anterolateral ligament (ALL) reconstruction was performed in this study with the aim to assess the clinical role of ALL in knee's stability and joint functions. Sixty patients were recruited and divided into three groups to perform the operations of anatomic single-bundle ACL reconstruction, anatomic double-bundle ACL reconstruction, and anatomic single-bundle ACL reconstruction + anterolateral ligament reconstruction, respectively. And then postoperative knee's stability and joint functions were evaluated to compare the clinical outcomes among the three different kind of operations. The postoperative knee's stability and joint functions of the anatomic double-bundle ACL reconstruction group and the anatomic single-bundle ACL reconstruction + ALL reconstruction group were better than the anatomic single-bundle ACL reconstruction group. No significant difference was observed between the anatomic double-bundle ACL reconstruction group and the anatomic single-bundle ACL reconstruction + ALL reconstruction group. The anatomic anterolateral ligament reconstruction could improve the clinical outcomes after patients performed the anatomic single-bundle anterior cruciate ligament reconstruction. This indicates that the anterolateral ligament plays a crucial role in knee's stability and joint function, especially the rotational stability.

A comparison of remifentanil parturient-controlled intravenous analgesia with epidural analgesia: a meta-analysis of randomized controlled trials.

BACKGROUND: Epidural analgesia is generally accepted as the most effective form of pain relief during labor. Remifentanil patient-controlled IV analgesia (PCIA), which is less invasive than epidural analgesia, may be an attractive alternative. In this meta-analysis, we compared the efficacy and safety of the 2 analgesic techniques for labor pain. METHODS: Databases of PubMed, EMBASE, and Cochrane Library were searched independently by 2 reviewers to retrieve eligible randomized controlled clinical trials. The primary end points were pain scores at 1 and 2 hours, and the secondary end points were nausea, vomiting, pruritus, and umbilical artery pH values. Mean difference (MD) or risk ratio with 95% confidence intervals (CIs) were calculated for each end point. GRADE profiler was applied to assess the quality of evidence. RESULTS: Five eligible trials were retrieved and analyzed. We found that parturients with remifentanil PCIA had higher visual analog scale (10-cm scale) pain scores than those who received epidural analgesia at 1 hour (MD = 1.9 cm; 95% CI, 0.5-3.3; I = 94%) and 2 hours (MD = 3.0 cm; 95% CI, 0.7-5.2; I = 89%) after initiation of analgesia. There was no statistical difference between epidural analgesia and remifentanil PCIA in the incidence of nausea, vomiting, pruritus, or umbilical artery pH values. However, the CIs are quite wide and contain clinically significant differences. According to GRADE profiler, most end points had moderate quality except that pain scores at 1 hour were of low quality. CONCLUSIONS: This meta-analysis suggests that remifentanil PCIA is not superior to epidural analgesia in analgesic efficacy during labor. Given the wide CIs of the pooled results for secondary maternal and neonatal outcomes, definite conclusions cannot be drawn for those outcomes. Further studies are still warranted to validate these conclusions.

Continuous irrigation and drainage for early postoperative deep wound infection after posterior instrumented spinal fusion.

STUDY DESIGN: A retrospective study of clinical cases. PURPOSE: To evaluate the efficacy of continuous irrigation and drainage for early postoperative deep wound infection after posterior instrumented spinal fusion. SUMMARY OF BACKGROUND DATA: Aggressive debridement and irrigation has been recommended to treat postoperative wound infections after instrumented spinal fusion. However, this method of management, indicating repeating visits to the operating room until the wound is clean enough for closure, often results in prolonged hospitalization, increased cost, and sometimes compromise of the desired outcome. We hypothesize that repeat visits to the operating room for debridements can be avoided by aggressive debridements and primary closure with continuous irrigation and drainage for postoperative wound infections. METHODS: From 2004 to 2009, 23 patients with early postoperative deep wound infections after spinal fusion with instrumentation were surgically treated with thorough debridement and primary closure with continuous irrigation and drainage. All patients were followed up for 30.6 months (range, 24-54 mo). RESULTS: The mean duration of irrigation was 12.0 days (range, 7-16 d). In 21 patients (91.3%), the wound healed after continuous irrigation. The removal of the instrumentation or cages was not required in any case. Spinal fusion was achieved in all cases, except 1, where the patient developed a pseudoarthrosis at the L4-L5 level after L4-S1 fusion. The mean ODI for these 23 patients improved significantly from 53.4±18.7 preoperatively to 18.3±11.2 at the final follow-up visit (P<0.001). The mean JOA scores increased significantly from 15.5±4.1 preoperatively to 24.3±3.8 at the final follow-up (P<0.001). CONCLUSIONS: Continuous irrigation and drainage is an effective and safe method for the treatment of early postoperative deep wound infection after posterior instrumented spinal fusion.

[Microanalysis study of the inspirable nano-particles into lungs by SEM and XREDS].

Inspirable nano-particles into lungs in the atmosphere were studied in this paper. Field emission scanning electron microscope (FSEM) and X-ray energy dispersive spectrometer were used to investigate the morphology and major constituents of inspirable nano-particles into lungs systematically. The results showed that most of the inspirable nano-particles in the atmosphere are spherical and ellipsoidal, with smooth surface and dense structure. The smaller nano-particles are clustered into loose floccule, with the sizes in the range of 30 to 100 nm. The constant elements in the nano-particles are close consistent with the large particle pollutants, which mainly contain C, O, Al, Si, Na, Mg, K, Ca, Fe, S and Cl etc. The point Analysis of EDS confirmed that the element content of Cl and S in some nano-particles is significantly increased, while others mainly contain C and O. It is believed that the surface of nano inorganic dust particles was adsorbed by the organic pollutants to form the core-shell structure nano-particles pollutants in the process of aerosol formation. Thus, reducing anthropogenic emissions of organic pollutants has great influence on the formation of inspirable nano-particles.

Enhanced Iturin A Production of Engineered Bacillus amyloliquefaciens by Knockout of Endogenous Plasmid and Rap Phosphatase Genes.

Iturin A biosynthesis has garnered considerable interest, yet bottlenecks persist in its low productivity in wild strains and the ability to engineer Bacillus amyloliquefaciens producers. This study reveals that deleting the endogenous plasmid, plas1, from the wild-type B. amyloliquefaciens HM618 notably enhances iturin A synthesis, likely related to the effect of the Rap phosphatase gene within plas1. Furthermore, inactivating Rap phosphatase-related genes (rapC, rapF, and rapH) in the genome of the strain also improved the iturin A level and specific productivity while reducing cell growth. Strategic rap genes and plasmid elimination achieved a synergistic balance between cell growth and iturin A production. Engineered strain HM-DR13 exhibited an increase in iturin A level to 849.9 mg/L within 48 h, significantly shortening the production period. These insights underscore the critical roles of endogenous plasmids and Rap phosphatases in iturin A biosynthesis, presenting a novel engineering strategy to optimize iturin A production in B. amyloliquefaciens.

Improving salt-tolerant artificial consortium of Bacillus amyloliquefaciens for bioconverting food waste to lipopeptides.

High-salt content in food waste (FW) affects its resource utilization during biotransformation. In this study, adaptive laboratory evolution (ALE), gene editing, and artificial consortia were performed out to improve the salt-tolerance of Bacillus amyloliquefaciens for producing lipopeptide under FW and seawater. High-salt stress significantly decreased lipopeptide production in the B. amyloliquefaciens HM618 and ALE strains. The total lipopeptide production in the recombinant B. amyloliquefaciens HM-4KSMSO after overexpressing the ion transportor gene ktrA and proline transporter gene opuE and replacing the promoter of gene mrp was 1.34 times higher than that in the strain HM618 in medium containing 30 g/L NaCl. Lipopeptide production under salt-tolerant consortia containing two strains (HM-4KSMSO and Corynebacterium glutamicum) and three-strains (HM-4KSMSO, salt-tolerant C. glutamicum, and Yarrowia lipolytica) was 1.81- and 2.28-fold higher than that under pure culture in a medium containing FW or both FW and seawater, respectively. These findings provide a new strategy for using high-salt FW and seawater to produce value-added chemicals.

Enhancement of polymyxin B1 production by an artificial microbial consortium of Paenibacillus polymyxa and recombinant Corynebacterium glutamicum producing precursor amino acids.

Polymyxin B, produced by Paenibacillus polymyxa, is used as the last line of defense clinically. In this study, exogenous mixture of precursor amino acids increased the level and proportion of polymyxin B1 in the total of polymyxin B analogs of P. polymyxa CJX518-AC (PPAC) from 0.15 g/L and 61.8 % to 0.33 g/L and 79.9 %, respectively. The co-culture of strain PPAC and recombinant Corynebacterium glutamicum-leu01, which produces high levels of threonine, leucine, and isoleucine, increased polymyxin B1 production to 0.64 g/L. When strains PPAC and C. glu-leu01 simultaneously inoculated into an optimized medium with 20 g/L peptone, polymyxin B1 production was increased to 0.97 g/L. Furthermore, the polymyxin B1 production in the co-culture of strains PPAC and C. glu-leu01 increased to 2.21 g/L after optimized inoculation ratios and fermentation medium with 60 g/L peptone. This study provides a new strategy to improve polymyxin B1 production.

Long noncoding RNA: an emerging paradigm of cancer research.

Recent studies have demonstrated the importance of non-protein coding part of human genome in carcinogenesis and metastasis. Among numerous kinds of non-protein coding RNAs, long noncoding RNAs (lncRNAs) play a key regulatory role in cancer biology. LncRNAs are dysregulated in different kinds of cancer and the expression levels of certain lncRNAs are associated with recurrence, metastasis, and prognosis of cancer. It is also proved that overexpression of certain lncRNAs, behaving like oncogenes, can promote matrix invasion of cancer cells and tumor growth. In this review, we focus our attention on lncRNAs those have been validated in human cancer tissues to suggest reasonable strategies for future research. We introduce an update view of lncRNA, extract cancer-related lncRNAs from literature, and describe the known functions and possible underlying molecular mechanisms of some well investigated lncRNAs (MALAT1, HOX antisense intergenic RNA, and highly upregulated in hepatocellular cancer), as well as their current and potential future application in cancer diagnosis (PCA3) and treatment (H19).

miR-221/222: promising biomarkers for breast cancer.

MicroRNAs (miRNAs) are small noncoding RNAs of 19-25 nt that can regulate gene expression at a posttranscriptional level. Increasing evidence indicates that miRNAs participate in almost every step of cellular processes and are often aberrantly expressed in human cancer. miR-221 and miR-222 are two highly homologous miRNAs that always act as a gene cluster (miR-221/222) in cellular regulation and have extensively been studied in cancer network. Here, we review the role of miR-221/222 in breast cancer (BCa) development and progression: regulating proliferative signaling pathways, altering telomere and telomerase activity, avoiding cell death from tumor suppressors, autophagy and apoptosis, monitoring angiogenesis, supporting epithelial-mesenchymal transition, and even controlling cell-specific function within microenvironment. We consider that miR-221/222 act as promising biomarkers for BCa and they would offer a new way in molecular targeting cancer treatment.

Secoiridoid glycosides from the fruits of Ligustrum lucidum and their in vitro anti-inflammatory activity.

Seven new secoiridoid glycosides (1-7), together with a known analogue (8), were isolated from the fruits of Ligustrum lucidum. Their structures with absolute configurations were determined by HR-ESI-MS, 1D and 2D NMR, and electronic circular dichroism (ECD) spectroscopic analysis, as well as biogenetic consideration. Compounds 1 and 2 are the first examples of secoiridoid glycoside dimers featuring a rare rearranged oleoside-type secoiridoid moiety, and compounds 3-7 represent a new class of oleoside-type secoiridoid glycosides with unusual stereochemistry at C-1 position. A plausible biosynthetic pathway for this group of unusual secoiridoid glycosides was also proposed herein. In addition, the isolates were evaluated for their in vitro anti-inflammatory activity, and all tested compounds exhibited modest inhibitory effects against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophages.

Enhancing fengycin production in the co-culture of Bacillus subtilis and Corynebacterium glutamicum by engineering proline transporter.

Fengycin possesses antifungal activity but has limited application due to its low yields. Amino acid precursors play a crucial role in fengycin synthesis. Herein, the overexpression of alanine, isoleucine, and threonine transporter-related genes in Bacillus subtilis increased fengycin production by 34.06%, 46.66%, and 7.83%, respectively. Particularly, fengycin production in B. subtilis reached 871.86 mg/L with the addition of 8.0 g/L exogenous proline after enhancing the expression of the proline transport-related gene opuE. To overcome the metabolic burden caused by excessive enhancement of gene expression for supplying precursors, B. subtilis and Corynebacterium glutamicum which produced proline, were co-cultured, which further improved fengycin production. Fengycin production in the co-culture of B. subtilis and C. glutamicum in shake flasks reached 1554.74 mg/L after optimizing the inoculation time and ratio. The fengycin level in the fed-batch co-culture was 2309.96 mg/L in a 5.0-L bioreactor. These findings provide a new strategy for improving fengycin production.

Effect of respirator resistance on tolerant capacity during graded load exercise.

Respirator breathing resistance impacts performance of wearers during constant work load. However, it is less clear as to how breathing resistance affects the tolerant capacity of users during graded work load. The present study investigated the tolerant capacity of 8 individuals during incremental work load. The 8 subjects were required to wear two matched respirators (respirators I and II which were designed to have different breathing resistances and the same dead space) respectively on separate days and then work to end points. Minute ventilation (V(E)), breathing frequency (BF), oxygen consumption (VO(2)) and heart rate (HR) were recorded during exercise, while tolerant time, response time and breathing discomfort were measured at the end of each test trial. The test variables were compared between the two respirators by using matched-pairs t-test. The results showed that the tolerant time was significantly reduced for the respirator I with higher level of breathing resistance when compared with its counterpart with lower breathing resistance (respirator II) (P<0.05). The same changes occurred for response time. Results also showed a significant increase in V(E) and BF for respirator I wearers when the work load was above 125 W. The O(2) consumption was similar under the two breathing resistance conditions. These findings suggested that the respiratory resistance caused by self-contained breathing apparatus (SCBA) has an impact on the tolerant capacity of users.

Stage IV non-small cell lung cancer among young individuals: Incidence, presentations, and survival outcomes of conventional therapies.

Background: There is a paucity of data published on the clinicopathological features and prognosis of stage IV non-small cell lung cancer (NSCLC) patients aged ≤45 years. Herein, we evaluated a large clinical series in an effort to provide a clearer picture of this population. Methods: The least absolute shrinkage and selection operator (LASSO)-penalized Cox regression model was performed to identify prognostic factors for NSCLC among individuals aged ≤45 years. The Kaplan-Meier method with log-rank test was used to compare overall survival (OS) differences between groups. Competing risk analysis with the Fine-Gray test was used to analyze cancer-specific survival (CSS) differences. Propensity score matching (PSM) was used to minimize selection bias. Results: Incidence-rate analyses, including 588,680 NSCLC cases (stage IV, 233,881; age ≤ 45 years stage IV, 5,483; and age > 45 years stage IV, 228,398) from 2004 to 2015, showed that the incidence of stage IV NSCLC among young individuals decreased over the years. In comparative analyses of clinical features and survival outcomes, a total of 48,607 eligible stage IV cases (age ≤ 45 years stage IV, 1,390; age > 45 years stage IV, 47,217) were included. The results showed that although patients in the young cohort were more likely to be diagnosed at advanced stages, they were also more likely to receive aggressive treatments. In addition, the survival rates of the young patients were superior to those of the older patients both before and after PSM. Conclusions: Stage IV NSCLC patients aged ≤45 years comprise a relatively small but special NSCLC subgroup. Although this population had better survival outcomes than older patients, these patients deserve more attention due to their young age and the significant socioeconomic implications.

Lymphovascular invasion: A non-sized T descriptor for stage IA non-small cell lung cancer.

BACKGROUND: Lymphovascular invasion (LVI) has not been included in the tumor-node-metastasis (TNM) staging manual of non-small-cell lung cancer (NSCLC). We aimed to investigate the predictive value of LVI on stage IA NSCLC and proposed a method of incorporating LVI into the T category based on the latest TNM staging manual. METHODS: The least absolute shrinkage and selection operator (LASSO)-penalized Cox multivariable regression model was performed to identify prognostic factors. The Kaplan-Meier method was used to compare overall survival (OS) and disease-free survival (DFS) between groups. Propensity score matching (PSM) was used to minimize bias. RESULTS: A total of 1452 eligible stage I NSCLC cases (stage IA without LVI, 1022 cases; stage IA with LVI, 120 cases; stage IB, 310 cases) were included. LASSO-penalized multivariable Cox analysis revealed that LVI was an independent prognostic factor for both OS and DFS. Survival analysis demonstrated that the survivals of stage IA NSCLCs without LVI were better than those of stage IA with LVI and stage IB NSCLCs. In the matched cohort, the survivals of stage IA NSCLCs with LVI were comparable to those of stage IB NSCLCs. CONCLUSIONS: Stage IA NSCLCs with LVI and stage IB NSCLCs had similar survivals, and we proposed that LVI might be a non-sized T descriptor that upstaged stage IA diseases to stage IB.

[Short needling for knee osteoarthritis with blood stasis obstruction and its effect on serum inflammatory factors].

OBJECTIVE: To compare the clinical efficacy and its effect on serum levels of tumor necrosis factor α(TNF-α), interleukin 1ß(IL-1ß), interleukin 6 (IL-6) and prostaglandin E2 (PGE2) between short needling (close-to-bone needling) and conventional acupuncture for knee osteoarthritis (KOA) with blood stasis obstruction. METHODS: A total of 68 KOA patients with blood stasis obstruction were randomized into a short needling group (34 cases, 3 cases dropped off) and a conventional acupuncture group (34 cases, 3 cases dropped off). The same acupoints (Dubi [ST 35], Neixiyan [EX-LE 4], Binzhong [Extra], Liangqiu [ST 34], etc. on the affected side) were selected in the two groups. In the short needling group, short needling technique was adopted, the needles were slowly inserted and the needle bodies were shaken, thus gradually penetrated to the bone. In the conventional acupuncture group, conventional acupuncture was adopted, the needles were penetrated to the muscle. After qi-arrival, Dubi (ST 35) and Neixiyan (EX-LE 4), Zusanli (ST 36) and Liangqiu (ST 34) were connected with CMNS6-1 electronic acupuncture instrument, with disperse-dense wave, 2 Hz/10 Hz in frequency, the current intensity was based on patients' feeling, the needles were retained for 30 min, at the same time, the knee joint was irradiated for 30 min with a special electromagnetic wave apparatus in the two groups. Once every other day, 3 times a week for 4 weeks. Before and after treatment, the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) score, knee joint pain visual analogue scale (VAS) score, inflammatory response related indexes (serum TNF-a, IL-1ß, IL-6 and PGE2) and knee joint ultrasound were observed，and the clinical effect was evaluated in the two groups. RESULTS: After treatment，the pain, stiffness, function scores and total scores of WOMAC were decreased as compared with those before treatment in the two groups (P<0.05), except for the pain score, the changes of above scores in the short needling group were greater than the conventional acupuncture group (P<0.05). After treatment, the VAS scores, serum levels of TNF-a, IL-1ß, IL-6, PGE2 and knee joint synovium thickness, intra-articular effusion were decreased as compared with those before treatment in the two groups (P<0.05), the levels of TNF-a, IL-1ß, IL-6 in the short needling group were lower than the conventional acupuncture group (P<0.05). The total effective rate in the short needling group was 87.1% (27/31), which was superior to 83.9% (26/31) in the conventional acupuncture group (P<0.05). CONCLUSION: Short needling could improve the knee joint function, relieve the pain and inflammatory response, improve the knee joint synovium inflammatory response, reduce the knee joint intra-articular effusion for KOA patients, its effect is better than conventional acupuncture.