S100A4+ macrophages facilitate zika virus invasion and persistence in the seminiferous tubules via interferon-gamma mediation.

Testicular invasion and persistence are features of Zika virus (ZIKV), but their mechanisms are still unknown. Here, we showed that S100A4+ macrophages, a myeloid macrophage subpopulation with susceptibility to ZIKV infection, facilitated ZIKV invasion and persistence in the seminiferous tubules. In ZIKV-infected mice, S100A4+ macrophages were specifically recruited into the interstitial space of testes and differentiated into interferon-γ-expressing M1 macrophages. With interferon-γ mediation, S100A4+ macrophages down-regulated Claudin-1 expression and induced its redistribution from the cytosol to nucleus, thus increasing the permeability of the blood-testis barrier which facilitated S100A4+ macrophages invasion into the seminiferous tubules. Intraluminal S100A4+ macrophages were segregated from CD8+ T cells and consequently helped ZIKV evade cellular immunity. As a result, ZIKV continued to replicate in intraluminal S100A4+ macrophages even when the spermatogenic cells disappeared. Deficiencies in S100A4 or interferon-γ signaling both reduced ZIKV infection in the seminiferous tubules. These results demonstrated crucial roles of S100A4+ macrophages in ZIKV infection in testes.

Simple security proof of coherent-one-way quantum key distribution.

Coherent-one-way quantum key distribution (COW-QKD), which requires a simple experimental setup and has the ability to withstand photon-number-splitting attacks, has been not only experimentally implemented but also commercially applied. However, recent studies have shown that the current COW-QKD system is insecure and can only distribute secret keys safely within 20 km of the optical fiber length. In this study, we propose a practical implementation of COW-QKD by adding a two-pulse vacuum state as a new decoy sequence. This proposal maintains the original experimental setup as well as the simplicity of its implementation. Utilizing detailed observations on the monitoring line to provide an analytical upper bound on the phase error rate, we provide a high-performance COW-QKD asymptotically secure against coherent attacks. This ensures the availability of COW-QKD within 100 km and establishes theoretical foundations for further applications.

Secure and practical multiparty quantum digital signatures.

Quantum digital signatures (QDSs) promise information-theoretic security against repudiation and forgery of messages. Compared with currently existing three-party QDS protocols, multiparty protocols have unique advantages in the practical case of more than two receivers when sending a mass message. However, complex security analysis, numerous quantum channels and low data utilization efficiency make it intractable to expand three-party to multiparty scenario. Here, based on six-state non-orthogonal encoding protocol, we propose an effective multiparty QDS framework to overcome these difficulties. The number of quantum channels in our protocol only linearly depends on the number of users. The post-matching method is introduced to enhance data utilization efficiency and make it linearly scale with the probability of detection events even for five-party scenario. Our work compensates for the absence of practical multiparty protocols, which paves the way for future QDS networks.

Suppression of TCAB1 expression induced cellular senescence by lessening proteasomal degradation of p21 in cancer cells.

BACKGROUND: TCAB1, a.k.a. WRAP53ß or WDR79, is an important molecule for the maintenance of Cajal bodies and critically involved in telomere elongation and DNA repair. Upregulation of TCAB1 were discovered in a variety types of cancers. However, the function of TCAB1 in tumor cell senescence remains absent. METHODS: The TCAB1 knockdown cell lines were constructed. The expression levels of TCAB1, p21, p16 and p53 were detected by qRT-PCR and western blotting. Staining of senescence-associated ß-galactosidase was used to detect senescent cells. The ubiquitination of the p21 was analysed by immunoprecipitation and in vivo ubiquitination assay. TCGA databases were employed to perform in silico analyses for the mRNA expression of TCAB1, p21, p16 and p53. RESULTS: Here, we discovered that knockdown of TCAB1 induced rapid progression of cellular senescence in A549, H1299 and HeLa cells. In exploiting the mechanism underlining the role of TCAB1 on senescence, we found a significant increase of p21 at the protein levels upon TCAB1 depletion, whereas the p21 mRNA expression was not altered. We verified that TCAB1 knockdown was able to shunt p21 from proteasomal degradation by regulating the ubiquitination of p21. In rescue assays, it was demonstrated that decreasing the expression of p21 or increasing the expression of TCAB1 were able to attenuate the cellular senescence process induced by TCAB1 silencing. CONCLUSIONS: This study revealed the importance of TCAB1 for its biological functions in the regulation of cell senescence. Our results will be helpful to understand the mechanisms of senescence in cancer cells, which could provide clues for designing novel strategies for developing effective treatment regimens.

The prognostic potential of coilin in association with p27 expression in pediatric acute lymphoblastic leukemia for disease relapse.

BACKGROUND: Cajal body (CB) is a nucleic organelle where small nuclear ribonucleoproteins undergo modification, maturation, splicing and/or assembly. Coilin is the marker structural protein of CBs. The expression level and cellular localization of coilin is sensitive to chemotherapeutic reagents, such as cisplatin. The gene of cyclin-dependent kinase inhibitor 1B (p27) is located with a high incidence translocation region of leukemic chromosomes, and its expression was of prognosis values in a variety of adult leukemia types. The exact profile and associated functions of coilin, as well as p27, in children's acute lymphoblastic leukemia (ALL) is obscure. METHODS: Bone marrow samples from 144 patients with ALL were collected. The expression levels of coilin and p27 were detected by qRT-PCR. The patient cohort was divided into low and high groups of coilin and p27 respectively. The prognosis and clinicobiological characteristics of different groups were investigated, especially focused on the treatment outcome. Leukemia cells of Reh or RS4;11 were exposed to different concentrations of DNR, prior to the detection for morphological changes of coilin by immunofluorescence. In Reh cells, lentivirus sh-coilin was used to silence coilin expression. Western blotting was used to detect coilin and p27 expression; flow cytometry was used for cell cycle and apoptosis assay; MTS method was used for measuring cell viability to examine the drug sensitivity of DNR. RESULTS: In this study, we found that daunorubicin was able to induce significant morphological changes of CBs in Reh and RS4;11 cells. Knockdown the expression of coilin increased the sensitivity to daunorubicin and inhibited the expression of p27 in Reh cells, and led to increased apoptosis. Importantly, not only the levels of coilin and p27 mRNA expression at initial diagnosis ALL children are markedly higher than those at complete remission (CR), but also both coilin and p27 expression in the relapsed patients was observed significantly higher comparing to the continuous CR patients. The 4-year EFS and RFS indicated that low levels of both coilin and p27 group favored better prognosis (p < 0.05). CONCLUSIONS: Our results indicated that consideration of coilin and p27 levels could be a prognostic reference for predicting the outcome of pediatric ALL patients, especially for disease recurrence. Reduction of coilin expression was sufficient to increase the sensitivity of leukemic cells to daunorubicin treatments, and during which possibly involved functions of p27 in cell cycle regulation and its effects on cell apoptosis.

Reliable HPLC separation, vibrational circular dichroism spectra, and absolute configurations of isoborneol enantiomers.

Resolution of chiral compounds has played an important role in the pharmaceutical field, involving detailed studies of pharmacokinetics, physiological, toxicological, and metabolic activities of enantiomers. Herein, a reliable method by high-performance liquid chromatography (HPLC) coupled with an optical rotation detector was developed to separate isoborneol enantiomers. A cellulose tris(3, 5-dimethylphenylcarbamate)-coated chiral stationary phase showed the best separation performance for isoborneol enantiomers in the normal phase among four polysaccharide chiral packings. The effects of alcoholic modifiers and column temperature were studied in detail. Resolution of the isoborneol racemate displayed a downward trend along with an increase in the content of ethanol and column temperature, indicating that less ethanol in the mobile phase and lower temperature were favorable to this process. Moreover, two isoborneol enantiomers were obtained via a semipreparative chiral HPLC technique under optimum conditions, and further characterized by analytical HPLC, and experimental and calculated vibrational circular dichroism (VCD) spectroscopy, respectively. The solution VCD spectrum of the first-eluted component was consistent with the Density Functional Theory (DFT) calculated pattern based on the SSS configuration, indicating that this enantiomer should be (1S, 2S, 4S)-(+)-isoborneol. Briefly, these results have provided reliable information to establish a method for analysis, preparative separation, and absolute configuration of chiral compounds without typical chromophoric groups.

[Effect of Electroacupunctrue on ERp57 in NAFLD Rats].

OBJECTIVE: To investigate the effect of electroacupunctrue on ERp57 in non-alcoholic fatty liver disease (NAFLD) rats, and study the therapeutic mechanism of electroacupunctrue in patients with NAFLD. METHODS: Sixty male SD rats were randomly divided into common diet group (n = 15) and high-fat diet group (n = 45). 5 weeks later, two rats from the two groups were executed and confirmed that the model was successful. Then 10 rats in common diet group were chosen as control group (Control), and 40 rats in high-fat diet group were randomly chosen and divided into diet group 1 (D1), diet group 2 (D2), electroacupuncture group 1 (EA) and electroacupuncture group 2 (EA2) (n = 10 each). D1 and EA1 were fed by high fat diet; D2 and EA2 were fed with common diet. In EA1 and EA2, filiform needle acupuncture was applied to ST36, SP6 and Liv3 and electroacupunctrue was applied to one-side of ST36, SP6 for 20 min once daily for 4 weeks. The rats in each group were weighed per-week. After the treatment the changes of blood lipid and liver functions of these rats were observed. ERp57 gene expression and protein expression were detected by RT-PCR and Western blot, and expression of ERp57 downstream SREBP-1c was detected. RESULTS: The body mass of D1 increased slowly and were lower than D2 and EA1 (P < 0.05); the body weights of EA2 increased rapidly and were higher than EA1 (P < 0.05), but without significant difference with D2 (P > 0.05). The contents of blood lipid, liver functions and the expression of ERp57 and SREBP-1c were significantly higher than those in Control, D2 and EA1 (P < 0.05). While compared to D2 and EA1 respectively, the index mentioned above in EA2 decreased more significantly (P < 0.05). CONCLUSION: Electroacupunctrue can decrease expression of ERp57 to improve endoplasmic reticulum stress (ERS) of rats with NAFLD and then decrease expression of SREBP-1c to regulate rat lipid, which could be one of mechanism to cure NAFLD.

A prospective, single-centre, randomized, double-blind controlled study protocol to study whether long-term oral metronidazole can effectively reduce the incidence of postoperative liver metastasis in patients with colorectal cancer.

INTRODUCTION: Fifteen to 25% of patients with colorectal cancer have combined liver metastases at the time of diagnosis, whereas an additional 15 to 25% will develop liver metastases after curative resection of primary colorectal cancer, with the vast majority (80-90%) of liver metastases unresponsive to curative resection at first. Colorectal cancer liver metastasis is also the leading cause of death in patients with colorectal cancer. In recent years, several studies have demonstrated that intestinal flora, especially Fusobacterium nucleatum, plays a crucial role in the development of colorectal cancer liver metastasis, so we hypothesized that long-term metronidazole use could effectively reduce the incidence of postoperative liver metastasis in colorectal cancer patients. METHODS/DESIGN: This study is a prospective, single-centre, randomized, double-blind controlled study in which 300 patients will be randomly assigned to the test group or the control group in a 1:1 allocation ratio. The aim of this trial is to demonstrate that long-term oral antibiotics can effectively reduce the incidence of postoperative liver metastasis in patients with colorectal cancer. ETHICS AND DISSEMINATION: Ethics approval was obtained from the Ethics Committee at the Chinese Ethics Committee of Registering Clinical Trials (ChiECRCT20210229). The results of this study will be disseminated at several research conferences and as published articles in peer-reviewed journals. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2100046201. Registered on July 05, 2021.

Are preoperative oral antibiotics effective in reducing the incidence of anastomotic leakage after colorectal cancer surgery? Study protocol for a prospective, multicentre, randomized controlled study.

INTRODUCTION: The optimal preoperative preparation for elective colorectal cancer surgery has been debated in academic circles for decades. Previously, several expert teams have conducted studies on whether preoperative mechanical bowel preparation and oral antibiotics can effectively reduce the incidence of postoperative complications, such as surgical site infections and anastomotic leakage. Most of the results of these studies have suggested that preoperative mechanical bowel preparation for elective colon surgery has no significant effect on the occurrence of surgical site infections and anastomotic leakage. METHODS/DESIGN: This study will examine whether oral antibiotic bowel preparation (OABP) influences the incidence of anastomotic leakage after surgery in a prospective, multicentre, randomized controlled trial that will enrol 1500 patients who require colon surgery. The primary endpoint, incidence of anastomotic leakage, is based on 2.3% in the OABP ± mechanical bowel preparation (MBP) group in the study by Morris et al. Patients will be randomized (1:1) into two groups: the test group will be given antibiotics (both neomycin 1 g and metronidazole 0.9 g) the day before surgery, and the control group will not receive any special intestinal preparation before surgery, including oral antibiotics or mechanical intestinal preparation. All study-related clinical data, such as general patient information, past medical history, laboratory examination, imaging results, and surgery details, will be recorded before surgery and during the time of hospitalization. The occurrence of postoperative fistulas, including anastomotic leakage, will be recorded as the main severe postoperative adverse event and will represent the primary endpoint. ETHICS AND DISSEMINATION: Ethics approval was obtained from the Chinese Ethics Committee of Registering Clinical Trials (ChiECRCT20200173). The results of this study will be disseminated at several research conferences and as published articles in peer-reviewed journals. Protocol was revised on November 22, 2021, version 4.0. TRIAL REGISTRATION: ChiCTR2000035550 . Registered on 13 Aug 2020.

Study protocol for comparing the efficacy of left-open single-flap technique versus double-flap technique after proximal gastrectomy: A multicenter randomized controlled trial.

Background: Proximal gastrectomy has gradually gained more attention due to its superiority in retaining the function of part of the stomach. The inevitable loss of the antireflux barrier and postoperative complications resulting from proximal gastrectomy can severely affect the quality of life. Continuous improvements in digestive tract reconstruction after proximal gastrectomy have yielded the development of a variety of methods with antireflux functions. Recently, our center attempted the left-open single-flap technique and initiated a multicenter, prospective, randomized controlled trial for patients undergoing proximal gastrectomy to reduce the difficulty of surgical anastomosis and the incidence of perioperative complications compared with the double-flap technique. These findings will provide more evidence-based medical research for the development of clinical guidelines. Methods/design: This study is a prospective, multicenter, randomized controlled clinical trial. We plan to recruit 250 patients who are eligible for proximal gastrectomy. After informed consent is obtained, patients will be randomly assigned to the trial group (left-open single-flap technique) and the control group (double-flap technique) in a 1:1 allocation ratio. Discussion: Increasingly, clinical studies have focused on the improvement of reconstruction modalities after proximal gastrectomy. Among these methods, the double-flap technique is a clinically effective method. The purpose of this study is to establish a prospective randomized controlled trial to compare the efficacy of the left-open single-flap technique versus the double-flap technique after proximal gastrectomy, aiming to provide more evidence-based medical studies for digestive tract reconstruction in proximal gastrectomy. Clinical Trial Registration: ClinicalTrials.gov, identifier [NCT05418920].

Protocol for Comparing the Efficacy of Three Reconstruction Methods of the Digestive Tract (Kamikawa Versus Double-Tract Reconstruction Versus Tube-Like Stomach) After Proximal Gastrectomy.

Background: Appropriate gastrointestinal reconstruction after proximal gastrectomy can effectively reduce the incidence of postoperative complications in patients with proximal early gastric cancer. However, there is still great controversy about the choice of digestive tract reconstruction after proximal gastrectomy, and there is no clinical consensus on the choice of digestive tract reconstruction after proximal gastrectomy. Currently, there is a lack of large-sample, prospective, randomized controlled studies to compare the efficacy of Kamikawa, double-tract reconstruction, and tube-like stomach reconstruction after proximal gastrectomy. Methods/design: This study will investigate the efficacy of three reconstruction methods after proximal gastrectomy in a prospective, multicenter, randomized controlled trial, which will enroll 180 patients with proximal early gastric cancer. Patients will be randomly divided into three groups: Group A (Kamikawa, n = 60), Group B (double-tract reconstruction, n = 60), and Group C (tube-like stomach, n = 60). The general information, past medical history, laboratory findings, imaging findings, and surgical procedures of the patients will be recorded and analyzed. The incidence of reflux esophagitis will be recorded as the primary endpoint. The incidence of anastomotic leakage, anastomotic stenosis, operative time and intraoperative blood loss will be recorded as secondary endpoints. Discussion: This study will establish a large-sample, prospective, randomized controlled trial to compare the efficacy of Kamikawa, double-tract reconstruction, and tube-like stomach reconstruction after proximal gastrectomy. Trial registration: This study was approved by the Chinese Clinical Trial Registry and registered on April 30, 2021. The registration number is ChiCTR2100045975.

A retrospective study of the role of preoperative ultrasonography in the detection of deep vein thrombosis in 1750 patients with gastric and colorectal cancers.

BACKGROUND: There has been a lack of research in the past on the prevalence and risk factors associated with deep vein thrombosis (DVT) in patients with resectable gastric and colorectal cancers. The purpose of this study was to review the anatomical distribution, prevalence and risk factors associated with lower limb DVT in 1750 patients with preoperative gastric and colorectal cancers and to evaluate the role of preoperative ultrasonography in the detection of DVT in preventing postoperative pulmonary thromboembolism (PTE) in patients with gastric and colorectal cancers. METHODS: A total of 1750 patients with gastric and colorectal cancers who underwent preoperative venous ultrasonography of the lower limbs were retrospectively reviewed. The risk factors associated with preoperative DVT were identified using univariate and multivariate logistic regression analysis. RESULTS: Seventy-three of the 1750 patients with gastric and colorectal cancers had DVT detected by preoperative venous ultrasonography of the lower limb and the incidence of lower limb DVT was 4.17 % in 1750 patients with gastric and colorectal cancers. Univariate analysis showed a higher risk of DVT in patients who met the following criteria: aged ≥80 years, female sex, the performance status ≥1, stage IV, ASA class ≥ III/IV, and hypertension. Multivariate logistic regression analysis showed that female sex, stage IV and ASA class ≥ III/IV were significantly associated with DVT before gastric and colorectal cancer surgery. CONCLUSIONS: Our study showed that female sex, stage IV and ASA class ≥ III/IV were significantly associated with DVT before gastric and colorectal cancer surgery. Routine venous ultrasonography for the lower limb can identify the risk of PTE, which is of great significance in the prevention and occurrence of PTE.

[Changes in expression of autophagy-related factors during acute contusion repair of skeletal muscle].

OBJECTIVES: To explore the possible biological mechanism of skeletal muscle contusion repair through researching the changes in expression of autophagy-related genes and proteins in SD rats with acute skeletal muscle contusion. METHODS: Six rats were randomly selected as the control group from 30 male SD rats, acute skeletal muscle contusion model were established in the remaining 24 rats with self-made hitter, then the model rats were randomly divided into 4 groups (3 d, 5 d, 7 d, 14 d groups, n=6). On the 3rd, 5th, 7th and 14th day after injury, injured gastrocnemius of each group was harvested. The morphological and the ultra-microstructure changes of gastrocnemius after injury were observed by HE staining and transmission electron microscope (TEM) respectively. The relative protein levels of (LC3-Ⅱ) and P62 of each group were observed by Western blot. The relative mRNA levels of atg7, atg10, atg12, atg16L1 of each group were observed by RTPCR. RESULTS: The results of HE staining showed that compared with the control group, the inflammation reached its peak on the 5th day after injury, new muscle fibers were clearly observed in 7 d group. The results of TEM showed that, compared with the control group, oncotic mitochondria could be clearly seen in the 3 d, 5 d, 7 d groups. Also, the Z line changed from disappearing to drift thickening, sarcoplasmic reticulum dilatation gradually improved, there was no evident difference between the 14 d group and the control group, suggesting that the damage has preliminarily healed. The results of Western blot showed that the expressions of LC3-Ⅱand P62 were increased at first and then decreased. The expression of LC3-Ⅱwas markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0.01). Similarly, compared with the control group, the expression of P62 reached its peak on the 3rd day after injury (P<0. 01), and returned to normal level on the 14th day. The results of RT-PCR showed that the expression of atg10 mRNA in the natural recovery group of 3 d, 5 d, 7 d, 14 d was firstly decreased and then increased, the atg10 mRNA was markedly down-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0. 01). The expression of atg7, atg12, atg16L1 mRNA was generally increased at first and then decreased, it was markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0.01, P<0.05, P<0.01). CONCLUSIONS: The above results indicate that the autophagy is involved in repair of skeletal muscle injury by its autophagyrelated factors,regularly changes after contusion, and the rate of damage repair may be related to the level of autophagy.

[Effect of Electroacupunctrue Stimulation at "Fenglong"(ST 40) on Expression of SREBP-1 c in Non-alcoholic Fatty Liver Disease Rats].

OBJECTIVE: To observe the effect of electroacupuncture(EA) stimulation at "Fenglong"(ST 40) on expression of sterol regulatory element binding protein-1 c(SREBP-1 c) in non-alcoholic fatty liver disease(NAFLD) rats, so as to reveal its mechanism underlying improving NAFLD. METHODS: Male SD rats were randomly divided into normal diet group, high-fat diet group, ST 40 manual acupuncture (MA) group, ST 40 EA group and "Zusanli"(ST 36) EA group(n=12 in each group). Apart from the normal diet group, rats in other groups were fed with high-fat diet to replicate NAFLD model. ST 40 MA group was treated with filiform needling for 20 min, EA (4 Hz/20 Hz, 5 mA) was applied to bilateral acupoints of ST 40 and ST 36 respectively for 20 min in the ST 40 EA group and ST 36 EA group, once a day for 4 weeks. The pathological changes of hepatic tissue were observed by HE staining. The contents of serum free fatty acid (FFA), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by colorimetry. The expression of SREBP-1 c mRNA and protein in the hepatic tissue were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot, respectively. RESULTS: HE staining showed that hepatocytes disorganized with diffuse bullous fat vacuolar changes in rats of the high-fat diet group, and small vesicle steatosis appeared in the ST 40 MA group. The liver cells arranged orderly and the steatosis decreased in the ST 40 EA group and ST 36 EA group. The contents of FFA, ALT, AST in serum and the expression levels of SREBP-1 c mRNA and protein in the hepatic tissue were significantly higher in the high-fat diet group than in the normal diet group(P<0.01), while significantly lower in the MA and EA groups than in the high-fat diet group(P<0.01). Furthermore, the effects of treatment in the ST 40 EA group were better than those in the ST 40 MA group(P<0.01, P<0.05). The FFA contents in serum and the expression levels of SREBP-1 c mRNA and protein were decreased in the ST 40 EA group in comparison with the ST 36 EA group(P<0.05). CONCLUSIONS: EA at ST 40 and ST 36 have a positive regulating effect on NAFLD rats, which may be related to down-regulating the expression of SREBP-1 c mRNA and protein in hepatic tissue, improving the endoplasmic reticulum stress and regulating lipid metabolic disorder, so as to relieve the inflammatory injury in hepatic tissue, and ST 40 works better.

[Effects of Electroacupuncture on Adaptive Hypertrophy of Gastrocnemius and Proliferation and Differentiation of Muscle Satellite Cells in Rats].

OBJECTIVE: To explore the effects and mechanism of different duration electroacupuncture (EA) on adaptive hypertrophy of gastrocnemius and the expressions of proliferation cell nuclear antigen (PCNA), paired box transcription factor 7 (Pax 7), myogenic differentiation antigen (MyoD 1), myogenin (MyoG), myosin heavy chain-Ⅰ(Myh 7), transforming growth factor-ß 1 (TGF-ß 1), Smad 3 in rats. METHODS: Twenty-four SD adult male rats were randomly divided into normal, two-week EA (A 2), four-week EA (A 4) and eight-week EA (A 8) groups, 6 rats in each group. Except the normal group, EA was applied at right "Zusanli" (ST 36) and "Huantiao" (GB 30) for 10 min in the other three groups, separately for 2 weeks, 4 weeks and 8 weeks, once a day, 6 times a week. The ratio of gastrocnemius wet weight was calculated after EA. The cross-sectional area and diameter of muscle fiber were measured after HE staining. The expression of PCNA protein was detected by immunofluorescence. The relative expressions of Pax 7, MyoD 1, MyoG, Myh 7, TGF-ß 1 and Smad 3 mRNA were tested with real-time PCR. RESULTS: The expressions of PCNA protein were obviously higher in the A 2 and A 4 groups than that in the normal group. The expression of genes in the EA intervention groups showed a trend of increasing first and then decreasing except Smad 3 mRNA. The relative expressions of Pax 7 and TGF-ß 1 mRNAs reached their peaks in the A 2 group compared with those in the normal group (P<0.01). The ratio of gastrocnemius wet weight, cross-sectional area and diameter of muscle fiber, and the relative expressions of MyoD 1, MyoG, Myh 7 mRNAs reached their peaks in the A 4 group (P<0.01), while the relative expression of Smad 3 mRNA showed a trend of decreasing first and then increasing and reached its bottom at the 4th week (P<0.01). CONCLUSIONS: EA at "Zusanli" (ST 36) and "Huantiao" (GB 30) may induce adaptive hypertrophy of gastrocnemius in rats by up-regulating the expressions of Pax 7, PCNA, TGF-ß 1, MyoD 1, MyoG, Myh 7 and down-regulating the expression of Smad 3, promoting the proliferation of skeletal satellite cells, myogenic differentiation, myotubes terminal differentiation, increasing the wet weight, fiber cross-sectional area and diameter of gastrocnemius.

[The Effect of Electroacupuncture Combined with Treadmill Training on Skeletal Muscle Contusion in Rats].

OBJECTIVE: To study the effect and mechanism of electroacupuncture(EA)combined with treadmill training in rats with skeletal muscle contusion. METHODS: A total of sixty Sprague Dawley rats were randomly divided into normal,model, EA, treadmill, and combination groups (n=12/group). The self-made striking device was used to establish skeletal muscle contusion model. EA was applied at "Zusanli" (ST 36) and Ashi point in the EA group. Treadmill was applied to train rats in the treadmill group. Rats in the combination group were received the above two methods. All the treatment was given once a day until the 3rd and 14th days after injury. The cross-sectional area and diameter of neonatal gastrocnemius muscle cells 14 days after injury were observed by HE staining. The expression levels of mammalian target of rapamycil (mTOR), myogenin (myoG) in gastrocnemius 3 days after injury and gastrocnemius mTOR, Fast myosin skeletal heavy chain (Fast MyHC) 14 days after injury were observed by Western blot. RESULTS: The cross-sectional area and diameter of neonatal muscle gastrocnemius cells in the model group were significantly lower than those in the normal group (P<0.01), and those in all the intervention groups were significantly higher than those of the model group (P<0.05), with better results in the combination group compared with those in the EA and treadmill groups (P<0.05). On the 3rd day after injury, the expressions of mTOR and myoG proteins in the model group were significantly higher compared with those in the normal group (P<0.01), and those in all the intervention groups were up-regulated in comparison with the model group (all P<0.01). The mTOR and myoG proteins in the treadmill group were lower than those in the EA group (P<0.01). The above two indexes in the combination group were higher than those in the EA and treadmill groups (P<0.05, P<0.01). On the 14th day after injury, the expression of mTOR was up-regulated in the model group compared with that in the normal group, but without significant difference (P<0.05), while the expression of Fast MyHC decreased (P<0.01). The expressions of mTOR and Fast MyHC in all the intervention groups increased compared with those in the model group (P<0.05, P<0.01). The expressions of mTOR and Fast MyHC proteins in the treadmill group were significantly down-regulated compared with those in the EA group (P<0.05, P<0.01). The two indexes in the combination group were higher than those in the other two intervention groups (P<0.01). CONCLUSIONS: EA combined with treadmill training can improve the myogenic differentiation and maturation, alleviate the injury of skeletal muscle, which may be related to its effect of increasing the expression of mTOR protein and positively regulating myoG and Fast MyHC proteins.

[Effects of Electroacupuncture on Apoptosis-related Protein Expression of Gastrocnemius Muscle Cells in Rats with Denervated Sciatic Nerve].

OBJECTIVE: To observe the anti-apoptosis effect of electroacupuncture (EA) on gastrocnemius muscle cells in rats with denervated sciatic nerve, so as to explore its possible mechanisms underlying delaying atrophy of skeletal muscle. METHODS: Sixty-three male Sprague-Dawley rats were randomly divided into sham operation group, model group, and EA group, and then further divided into 3 subgroups in each group(n=7/subgroup). Gastrocnemius muscle atrophy model was established by transecting the sciatic nerve of rat. EA (5 Hz, 1.5 mA) was applied to "Zusanli" (ST 36) and "Chengshan" (BL 57) acupoints at the affected side for 10 min, once a day for 1, 2, 3 weeks, respectively. The gastrocnemius muscles were sampled on the 7th, 14th, 21th d after modeling, separately. The apoptotic cells were detected by TUNEL staining. Bcl-2, Bax, Cyt-C and Caspase-3 protein expressions were determined by Western blot. RT-PCR was used to check Caspase-3 gene expression. RESULTS: Compared with the sham operation group, the cell apoptotic index in gastrocnemius was markedly higher, gastrocnemius Bcl-2 protein expression was markedly down-regulated, and gastrocnemius Bax, Cyt-C and Caspase-3 protein expressions were considerably up-regulated in the model group at all time-points (P<0.05). Compared with the model group, the cell apoptotic indexes in gastrocnemius were significantly lower in the three EA subgroups (P<0.05). Bcl-2 protein expressions were markedly up-regulated while Bax, Cyt-C and Caspase-3 protein expressions were significantly down-regulated in the EA group 14, 21 days after modeling compared with the corresponding model subgroups (P<0.05). The changes of Caspase-3 mRNA expression levels of gastrocnemius in all groups were similar to those of Caspase-3 protein expressions. CONCLUSIONS: Electroacupuncture intervention can effectively increase Bcl-2 expression and decrease Bax, Cyt-C and Caspase-3 expression in gastrocnemius muscle, and consequently reduce the apoptosis of muscle cell, which may contribute to its effect in delaying the skeletal muscle atrophy of denervated sciatic nerve.

[Effects of Electroacupunctrue Combined with Dietary Control on Peroxisome Proliferator-activa- ted Receptor-α, and Liver Fatty Acid-binding Protein Levels in Non-alcoholic Fatty Liver Disease Rats].

OBJECTIVE: To observe the effect of electroacupunctrue (EA) intervention or EA combined with dietary control on peroxisome proliferator-activated receptor (PPAR)-α, and liver fatty acid-binding protein (L-FABP) levels in non-alcoholic fatty liver disease (NAFLD) rats, so as to reveal its mechanism underlying improvement of NAFLD. METHODS: Sixty SD male rats were randomly divided into common diet (control) group (n = 10) and high-fat diet group (n = 45). The NAFLD model was established by feeding the animals with high-fat forage (HFF, including cholesterol, sodium cholate, propylthiouracil, sucrose, lard and common forage) for 5 weeks. Forty NAFLD rats were then randomized into model, EA + HFF, low-fat forage (LFF) and EA+ LFF groups (n = 10 rats in each group). EA (4 Hz/20 Hz, 3 mA) was applied to ipsilateral "Zusanli" (ST 36),"Sanyinjiao" (SP 6) and "Taichong" (LR 3) for 20 min, once daily for 4 weeks. The pathologic changes of the hepatic tissue were detected by H. E. staining. Serum total cholesterol (TC) and triglyceride (TG) contents were determined by using enzymatic methods, serum free fat acids (FFA) content was detected by colorimetry. The expression levels of PPAR-α and L-FABP protein and gene of the liver tissue were determined by Western blot and RT-PCR, respectively. RESULTS: H. E. staining showed that the hepatocytes presented moderate or severe bullous adipose degeneration in rats of the model group, vesicular steatosis in the EA + HFF and LFF groups, turned to almost normal but with small amount of lipid droplets in the EA + LFF group. The contents of serum TC, TG and FFA were significantly higher in the model group than in the control group (P < 0.05), and were obviously decreased in the EA + HFF, LFF and EA + LFF groups in comparison with the model group (P < 0.05). Compared to the control group, hepatic PPAR-α protein and mRNA were markedly down-regulated in the model group, and hepatic L-FABP protein and mRNA considerably up-regulated in the model group (P < 0.05). Following EA intervention and low fat diet feeding, PPAR-α protein and mRNA were markedly up-regulated while L-FABP protein and mRNA remarkably down-regulated in the EA + HFF, LFF and EA + LFF groups (P < 0.05). CONCLUSION: EA intervention or EA combined with dietary control can relieve hepatic pathological changes in NAFLD rats, which may be related to its effects in down-regulating blood lipid level and hepatic L-FABP protein and mRNA expression and in up-regulating PPAR-α protein and mRNA expression, and the effect of combined treatment was relatively better than simple EA or LFF treatment.