Effects of ankle Kinesio taping on knee and ankle joint biomechanics during unanticipated jumps in collegiate athletes.

OBJECTIVE: Most biomechanical research on the application of Kinesio taping (KT) to the ankle joint focused on testing anticipated movements. However, ankle sprains frequently occur in real life in unanticipated situations, where individuals are unprepared and face sudden external stimuli. This situation is completely different from the anticipated situation. The aim of the present study was to investigate the effects of ankle KT application on the kinematic and kinetic characteristics of the knee and ankle joints during unanticipated jump tasks in collegiate athletes. METHODS: Eighteen healthy collegiate athletes experienced three taping conditions in a randomized order: no taping (NT), placebo taping (PT), and KT, and performed unanticipated jump tasks. A 9-camera infrared high-speed motion capture system was employed to collect knee and ankle kinematic data, and a 3-dimensional force plate was utilized to collect knee and ankle kinetic data during the tasks. RESULTS: During the right jumps, KT significantly increased peak knee flexion angle (P = 0.031) compared to NT and significantly decreased peak vertical ground reaction force (P < 0.001, P = 0.001) compared to NT and PT. During the left jumps, KT significantly reduced peak ankle inversion angle (P = 0.022, P < 0.001) and peak ankle inversion moment (P = 0.002, P = 0.001) compared to NT and PT. CONCLUSION: During unanticipated jump maneuvers, KT reduced peak ankle inversion angle, peak vertical ground reaction force, and peak ankle inversion moment and increased peak knee flexion angle in collegiate athletes.

Effects of Kinesio taping on lower limb biomechanical characteristics during the cutting maneuver in athletes after anterior cruciate ligament reconstruction.

PURPOSE: To determine the effects of Kinesio taping (KT) on the biomechanical characteristics of the lower limbs during the 90° cutting maneuver in anterior cruciate ligament (ACL) reconstruction (ACLR) athletes. METHOD: Eighteen ACLR athletes were recruited and subjected randomly to three taping conditions, KT, placebo taping (PT), and no taping (NT), followed by a 90° cutting test. A nine-camera infrared high-speed motion capture system (Vicon, T40, 200 Hz) was used to record the kinematic parameters of the lower limbs during the cutting maneuver, and a three-dimensional dynamometer (Kistler, 1000 Hz) was used to record the kinetic parameters of the lower limbs. A one-way repeated measures analysis of variance was conducted to compare the differences in the lower limb kinematic and kinetic characteristics of ACLR athletes subjected to these interventions. RESULTS: During the landing phase, the knee valgus angle reduced significantly with KT than with NT (95% confidence interval = -1.399 to -0.154; P = 0.025), whereas no significant difference was observed between PT and NT (95% confidence interval = -1.251 to 0.217; P = 0.236). No significant differences were observed in the other kinematic variables among the three taping conditions (P > 0.05). During the landing phase, no significant differences in the kinetic variables were observed among the three taping conditions (P > 0.05). CONCLUSIONS: Although KT does not improve the kinetic variables of athletes after ACLR during the 90° cutting maneuver, it reduces the knee valgus angle, which could reduce the risk of secondary ACL injury.

Abnormal Lower Limb Biomechanics During a Bilateral Vertical Jump Despite the Symmetry in Single-Leg Vertical Hop Height in Athletes After ACL Reconstruction.

Background: A limb symmetry index (LSI) of >90% for single-leg horizontal hop distance is recommended as a cutoff point for safe return to sports after anterior cruciate ligament reconstruction (ACLR). Despite achieving this threshold, abnormal lower limb biomechanics continue to persist in athletes after ACLR. Symmetry in single-leg vertical hop height appears to be more difficult to achieve and can be a better representation of knee function than single-leg horizontal hop distance. Purpose: To explore whether an LSI of >90% for single-leg vertical hop height can represent normal lower limb biomechanics in athletes during a bilateral vertical jump after ACLR. Study Design: Controlled laboratory study. Methods: According to the LSI for single-leg vertical hop height, 46 athletes who had undergone ACLR with an autologous ipsilateral bone-patellar tendon-bone or hamstring tendon graft were divided into a low symmetry group (LSI <90%; n = 23) and a high symmetry group (LSI >90%; n = 23), and 24 noninjured athletes were selected as the control group. The kinematic and kinetic characteristics during a bilateral vertical jump were compared between the low symmetry, high symmetry, and control groups. Results: During the propulsion phase of the bilateral vertical jump, the operated side in the high symmetry group showed a lower knee extension moment than the nonoperated side (P = .001). At peak vertical ground-reaction force, the operated side in the high symmetry group showed a lower knee internal rotation moment compared with the control group (P = .016). Compared with the nonoperated side, the operated side in the high symmetry group showed a higher hip extension moment (P = .002), lower knee extension moment (P < .001), lower ankle plantarflexion moment (P < .001), and lower vertical ground-reaction force (P = .023). Conclusion: Despite achieving an LSI of >90% for single-leg vertical hop height, athletes after ACLR showed abnormal lower limb biomechanical characteristics during the bilateral vertical jump. Clinical Relevance: Symmetrical single-leg vertical hop height may not signify ideal biomechanical or return-to-sports readiness in this population.

Effect of Platelet-Rich Plasma at Different Initiation Times on Healing of the Bone-Tendon Interface of the Rotator Cuff in a Mouse Model.

Background: Platelet-rich plasma (PRP) has demonstrated beneficial effects on healing of the bone-tendon interface (BTI). Purpose: To determine the optimal initiation time for PRP application after rotator cuff repair in an animal model. Study Design: Controlled laboratory study. Methods: A total of 136 C57BL/6 mice were included; 40 mice were used to prepare PRP, while 96 mice underwent acute supraspinatus tendon (SST) repair. The animals were randomly divided into 4 groups: a control group and 3 groups in which PRP was injected into the injury interface immediately after surgery, on the 7th postoperative day (PRP-7d), and on the 14th postoperative day. At 4 and 8 weeks postoperatively, the animals were sacrificed, blood was collected by eyeball removal, and samples of the SST-humerus complex were collected. Histological, imaging, immunological, and biomechanical data were compared among the groups using 1-way analysis of variance with the Bonferroni post hoc test. Results: Histological analysis revealed that the fibrocartilage layer at the BTI was larger in the PRP-7d group compared to the other groups at both 4 and 8 weeks postoperatively. Moreover, the PRP-7d group exhibited improved proteoglycan content and distribution compared to the other groups. Enzyme-linked immunosorbent assay results demonstrated that at 4 weeks postoperatively, higher concentrations of transforming growth factor-ß1 and platelet-derived growth factor-BB (PDGF-BB) were seen in the PRP-7d group versus the PRP-14d and control gruops (P < .05), and at 8 weeks postoperatively, the concentration of PDGF-BB was higher in the PRP-7d group versus the control group (P < .05). Biomechanical testing at 4 weeks postoperatively revealed that the failure load and ultimate strength of the SST-humerus complex were superior in the PRP-7d group compared to the other groups (P < .05), at 8 weeks, PRP-7d group was superior to the control group (P < .05). Additionally, at 8 weeks postoperatively, the PRP-7d group exhibited a greater trabecular number and trabecular thickness at the BTI compared to the PRP-14d and control gruops (P < .05). Conclusion: PRP promoted healing of the BTI after a rotator cuff injury at an early stage. Clinical Relevance: A PRP injection on the 7th postoperative day demonstrated superior therapeutic effects compared with injections at other time points.

Effects of Kinesio taping on lower limb biomechanical characteristics during unexpected jumping in patients with chronic ankle instability.

PURPOSE: The current biomechanical research on the application of Kinesio taping (KT) to patients with chronic ankle instability (CAI) has focused on testing the expected movements. However, unexpected movements are more common in actual sports. Therefore, the present study aimed to investigate the effects of KT on the biomechanical characteristics of the knee and ankle joints during unexpected jumping movements. METHODS: Twenty-one patients with unilateral CAI were recruited to capture the biomechanical parameters during unexpected jumping movements under different interventions: no taping (NT), placebo taping (PT), and KT. A one-way repeated measures analysis of variance was used to compare the differences in knee and ankle biomechanical characteristics among patients with CAI between the three intervention conditions. RESULTS: At initial contact, the KT group demonstrated a significant decrease in ankle plantarflexion and knee flexion angles compared to the NT group (p < 0.05). At the early landing phase, the KT group had a significant increase in peak ankle dorsiflexion angle, peak ankle eversion angle, peak ankle dorsiflexion moment, and peak ankle eversion moment compared to the NT and PT groups (p < 0.05). Furthermore, the KT group had a significantly reduced peak knee flexion angle, peak knee eversion angle, and peak vertical ground reaction force (p < 0.05) compared to the NT and PT groups. CONCLUSION: KT significantly improves the sprain-prone touchdown posture of patients with CAI. And reducing the risk of ankle sprains during the early landing phase by promoting ankle dorsiflexion and eversion angles and moments.

Eccentric Contraction Enhances Healing of the Bone-Tendon Interface After Rotator Cuff Repair in Mice.

BACKGROUND: Various muscle contraction modalities have differing effects on the musculoskeletal system. To understand the magnitude of these effects, the authors investigated the effects of eccentric and concentric contractions on the bone-tendon interface after rotator cuff repair in mice. HYPOTHESIS: Eccentric contraction promotes healing of the bone-tendon interface after rotator cuff repair in mice better than other muscle contraction patterns. STUDY DESIGN: Controlled laboratory study. METHODS: The authors performed acute supraspinatus tendon repair of the right shoulder in 104 C57BL/6 mice. Animals were randomized into 4 groups postoperatively: control group (Con group), horizontal running group (Horz group), +15° uphill running group (Up group), and -15° downhill running group (Down group), with 26 animals in each group. At 4 and 8 weeks postoperatively, the authors removed the eyeball, collected blood samples, and extracted the supraspinatus tendon-humerus complex for histological, immunological, bone morphological, and biomechanical tests. RESULTS: At 4 and 8 weeks postoperatively, the Down group exhibited a better collagen cell arrangement and fibrocartilage layer than the other 3 groups. At 4 weeks postoperatively, anti-inflammatory macrophages (M2 macrophages) were observed at the repair site in all groups except for the Con group. At 8 weeks postoperatively, M2 macrophages were withdrawn from the tendon site in all groups. The transforming growth factor ß1 concentration in the Down group was greater than that in the other 3 groups at 4 weeks postoperatively, and it was higher than that in the Con group at 8 weeks postoperatively. The bone volume fraction, number of trabeculae, and thickness of trabeculae at the repair site in the Down group, as well as the ultimate strength and failure load in the biomechanical tests, were greater than those in the other 3 groups at 8 weeks postoperatively. CONCLUSION: Eccentric contraction promotes healing of the bone-tendon interface after rotator cuff repair in mice better than other muscle contraction patterns. CLINICAL RELEVANCE: After clinical rotator cuff repair, patients can be rehabilitated by eccentric training to speed up the functional recovery of the shoulder joint.

Comparison of the effects of platelet-rich plasma and corticosteroid injection in rotator cuff disease treatment: a systematic review and meta-analysis.

HYPOTHESIS: Platelet-rich plasma (PRP) and corticosteroids are used to treat rotator cuff diseases. However, few reviews have compared the effects of these 2 treatments. In this study, we compared the effects of PRP and corticosteroid injection on the prognosis of rotator cuff diseases. MATERIALS AND METHODS: According to the Cochrane Manual of Systematic Review of Interventions, the PubMed, Embase, and Cochrane databases were searched comprehensively. Two independent authors screened suitable studies and performed data extraction and risk of bias assessment. Only randomized controlled trials comparing the effects of PRP and corticosteroid in the treatment of rotator cuff injuries were included, as measured by clinical function and pain during different follow-up periods. RESULTS: Nine studies with 469 patients were included in this review. In short-term treatment, corticosteroids were superior to PRP in the improvement of Constant, Simple Shoulder Test, and American Shoulder and Elbow Surgeons scores (mean difference [MD] -5.08, 95% confidence interval [CI] -10.26, 0.06; P = .05 and MD -0.97, 95% CI -1.68, -0.07; P = .03 and MD -6.67, 95% CI -12.85, -0.49; P = .03, respectively). No statistically significant difference was observed between the 2 groups at midterm (P > .05), and the recovery of the Simple Shoulder Test and American Shoulder and Elbow Surgeons scores in PRP treatment was significantly better than that in corticosteroid treatment in the long term (MD: 1.21, 95% CI: 0.68, 1.74; P < .00001 and MD: 6.96, 95% CI: 3.90, 9.61; P < .00001, respectively). In pain reduction based on visual analog scale score, corticosteroids led to better pain reduction (MD: 0.84, 95% CI: 0.03, 1.64; P = .04), but no significant difference was observed in pain reduction between the 2 groups in the any term (P > .05). However, these differences did not reach the minimum clinically important difference. CONCLUSIONS: Current analysis showed that corticosteroids have better efficacy in short term, whereas PRP is more beneficial for long-term recovery. However, no difference was observed in the mid-term efficacy between the 2 groups. Randomized controlled trials with longer follow-up periods and larger sample sizes are also needed to determine the optimal treatment.

Uncut interposed jejunum pouch versus esophago-gastrostomy and double anastomoses of jejunum to the esophagus and residual stomach: An innovative method of digestive tract reconstruction following proximal gastrectomy.

AIM: An innovative method of digestive tract reconstruction following proximal gastrectomy, the uncut interposed jejunum pouch, esophagus and residual stomach double anastomosis(Uncut-D), was established in recent years. In order to fully clarify the superiority of the procedure, this study has conducted a systematic analysis and thorough discussion. METHODS: 118 patients with adenocarcinoma of the esophagogastric junction who underwent proximal gastrectomy were enrolled in this study. According to the methods of digestive tract reconstruction, these patients were divided into three groups: Uncut-D(n = 43), esophagogastrostomy (EG, n = 36), jejunal interposition (JI, n = 39).The preoperative indicators, surgical complications and related indicators of postoperative quality of life were analyzed. RESULTS: There were no significant differences in preoperative data among all groups (P > 0.05); The digestive tract reconstruction time in Uncut-D group was more than that in EG group, and less than that in JI group (P < 0.05). The incidence of esophageal anastomotic stenosis in Uncut-D group was significantly lower than that in EG group (P < 0.05); In Uncut-D group, the incidence of reflux esophagitis, postoperative nutrition index(PNI), weight recovery and Visick classification were significantly better than those in EG group (P < 0.05), furthermore, the incidence of delayed gastric emptying,PNI and weight recovery were better than those in JI group (P < 0.05). CONCLUSIONS: The Uncut-D procedure gave full play to jejunal continuity and the advantages of pouch, and played a valuable role in gastric and cardiac replacement, which significantly reduced long-term complications, improved postoperative nutritional status of patients and long-term quality of life.

Comprehensive Analysis of Genomic Alterations in Hepatoid Adenocarcinoma of the Stomach and Identification of Clinically Actionable Alterations.

Hepatoid adenocarcinoma of the stomach (HAS) is a rare malignancy with aggressive biological behavior. This study aimed to compare the genetic landscape of HAS with liver hepatocellular carcinoma (LIHC), gastric cancer (GC), and AFP-producing GC (AFPGC) and identify clinically actionable alterations. Thirty-eight cases of HAS were collected for whole-exome sequencing. Significantly mutated genes were identified. TP53 was the most frequently mutated gene (66%). Hypoxia, TNF-α/NFκB, mitotic spindle assembly, DNA repair, and p53 signaling pathways mutated frequently. Mutagenesis mechanisms in HAS were associated with spontaneous or enzymatic deamination of 5-methylcytosine to thymine and defective homologous recombination-related DNA damage repair. However, LIHC was characteristic of exposure to aflatoxin and aristolochic acid. The copy number variants (CNVs) in HAS was significantly different compared to LIHC, GC, and AFPGC. Aggressive behavior-related CNVs were identified, including local vascular invasion, advanced stages, and adverse prognosis. In 55.26% of HAS patients there existed at least one clinically actionable alteration, including ERBB2, FGFR1, CDK4, EGFR, MET, and MDM2 amplifications and BRCA1/2 mutations. MDM2 amplification with functional TP53 was detected in 5% of HAS patients, which was proved sensitive to MDM2 inhibitors. A total of 10.53% of HAS patients harbored TMB > 10 muts/Mb. These findings improve our understanding of the genomic features of HAS and provide potential therapeutic targets.

Effects of leukocyte-rich platelet-rich plasma and leukocyte-poor platelet-rich plasma on the healing of bone-tendon interface of rotator cuff in a mice model.

Platelet-rich plasma (PRP) is widely used clinically to treat tendon injuries, and often contains leukocytes. However, the debate regarding the concentration of leukocytes in PRP is still ongoing. This study aimed to evaluate the therapeutic effects of leukocyte-rich platelet-rich plasma (LR-PRP) and leukocyte-poor platelet-rich plasma (LP-PRP) on the healing of the bone-tendon interface (BTI) of the rotator cuff. A total of 102 C57BL/6 mice were used. Thirty mice were used to prepare the PRP, while 72 underwent acute supraspinatus tendon injury repair. The animals were then randomly assigned to three groups: LR-PRP, LP-PRP and control groups. The mice were euthanized at 4 and 8 weeks postoperatively, and histological, immunological and biomechanical analyses were performed. The histological results showed that the fusion effect at the bone-tendon interface at 4 and 8 weeks after surgery was greater in the PRP groups and significantly increased at 4 weeks; however, at 8 weeks, the area of the fibrocartilage layer in the LP-PRP group increased significantly. M2 macrophages were observed at the repaired insertion for all the groups at 4 weeks. At 8 weeks, M2 macrophages withdrew back to the tendon in the control group, but some M2 macrophages were retained at the repaired site in the LR-PRP and LP-PRP groups. Enzyme-linked immunoassay results showed that the concentrations of IL-1ß and TNF-α in the LR-PRP group were significantly higher than those in the other groups at 4 and 8 weeks, while the concentrations of IL-1ß and TNF-α in the LP-PRP group were significantly lower than those in the control group. The biomechanical properties of the BTI were significantly improved in the PRP group. Significantly higher failure load and ultimate strength were seen in the LR-PRP and LP-PRP groups than in the control group at 4 and 8 weeks postoperatively. Thus, LR-RPR can effectively enhance the early stage of bone-tendon interface healing after rotator cuff repair, and LP-PRP could enhance the later stages of healing after rotator cuff injury.

Effect of Mechanical Stimulation Combined With Platelet-Rich Plasma on Healing of the Rotator Cuff in a Murine Model.

BACKGROUND: Mechanical stimulation and platelet-rich plasma (PRP) have been shown to be beneficial for healing of the bone-tendon interface (BTI), but few studies have explored the efficacy of a combination of these applications. We investigated the effect of mechanical stimulation combined with PRP on rotator cuff repair in mice. HYPOTHESIS: Mechanical stimulation combined with PRP can enhance BTI healing in a murine model of rotator cuff repair. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 160 C57BL/6 mice were used. Overall, 40 mice were used to prepare PRP, while 120 mice underwent acute supraspinatus tendon (SST) repair. The animals were randomly assigned to 4 groups: control group, mechanical stimulation group, PRP group, and mechanical stimulation combined with PRP group (combination group). At 4 and 8 weeks postoperatively, animals were sacrificed, the eyeballs were removed to collect blood, and the SST-humeral complexes were collected. Histological, biomechanical, immunological, and bone morphometric tests were performed. RESULTS: Histologically, at 4 and 8 weeks after surgery, the area of the fibrocartilage layer at the BTI in the combination group was larger than in the other groups. The content and distribution of proteoglycans in this layer in the combination group were significantly greater than in the other groups. At 8 weeks postoperatively, trabecular number, and trabecular bone thickness of the subchondral bone area of interest at the BTI of the combination group were greater than those of the other groups, bone volume fraction of the combination group was greater than the control group. On biomechanical testing at 4 and 8 weeks after surgery, the failure load and ultimate strength of the SST-humeral complex in the combination group were higher than in the other groups. Enzyme-linked immunosorbent assay results showed that, at 4 weeks postoperatively, the serum concentrations of transforming growth factor beta 1 and platelet-derived growth factor (PDGF) in the combination group were significantly higher than in the other groups; at 8 weeks, the PDGF-AB concentration in the combination group was higher than in the control and mechanical stimulation groups. CONCLUSION: Mechanical stimulation combined with PRP can effectively promote the early stage of healing after a rotator cuff injury. CLINICAL RELEVANCE: These findings imply that mechanical stimulation combined with PRP can serve as a potential therapeutic strategy for rotator cuff healing.

Repeated lumps and infections: A case report on breast augmentation complications.

BACKGROUND: Polyacrylamide hydrogel (PAAG) injections were once common in breast augmentation and have been prohibited for augmentation mammaplasty in China since a large number of patients who underwent breast augmentation with PAAG injections have continued to seek medical advice as a result of related complications. Among all these complications, distant migration is relatively rare. CASE SUMMARY: A 49-year-old female presented at the hospital with a one-year history of a vulvar lump. The sonography of the lump showed several subcutaneous fluid-filled regions from the left vulva to the pubic symphysis, which suggested possible fat liquefaction. An enhanced magnetic resonance imaging (MRI) revealed a cystic area, which was considered a benign lesion. Intraoperative observations showed that the mass did not have an obvious capsule, the subcutaneous tissue presented as a cavity, and some yellow material came out of this cavity. A culture of the drainage did not show bacterial contamination. Histopathology revealed a foreign body granuloma. After resection and closed drainage, lumps were successively observed in the left lower abdomen and the bilateral hypochondriac region with infections. Sonography found that the hypoechoic areas in the bilateral hypochondriac region seemed continuous with deep in the breasts. The patient reported that she had undergone surgery with PAAG injections 20 years ago after she was repeatedly asked about her past history. Finally, a diagnosis of distant migration of PAAG was made. CONCLUSION: PAAG gel can migrate after long periods of time. A diagnosis should not be limited to the area where the symptom develops.

MicroRNA-186 affects the proliferation of tumor cells via yes-associated protein 1 in the occurrence and development of pancreatic cancer.

The present study aimed to determine the expression of microRNA (miRNA or miR)-186 in tumor tissues and peripheral blood of patients with pancreatic cancer (PC), as well as its mechanism of regulation. A total of 65 patients with PC who underwent surgery between June 2013 and October 2015 were included. In addition, 59 healthy subjects were recruited as controls. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of mRNA and miRNA. Western blotting and enzyme-linked immunosorbent assay were used to determine protein expression. Bioinformatics was employed for the prediction of the target gene of miR-186, whereas dual luciferase reporter assay was performed to identify whether miR-186 directly bound to YAP1 mRNA. Human pulmonary aortic endothelial cells (HPACs) were transfected with ago-miR-186. YAP1 expression in HPACs was silenced by siRNA. MTT assay was used to evaluate the viability of HPACs. YAP1 mRNA and protein expression levels were elevated in PC. In addition, expression levels of miR-186 in PC were downregulated. miR-186 regulated the expression of YAP1 by binding with the 3'-untranslated region of YAP1. Elevated expression of miR-186 inhibited the proliferation of HPACs by downregulating the expression of YAP1. Decreased expression of YAP1 by siRNA reduced the viability of HPACs. The present study demonstrates that YAP1 is upregulated in the tumor tissues and blood of PC patients, and this may be associated with the downregulation of miR-186. In addition, miR-186 may affect the occurrence and development of PC by controlling the proliferation of PC cells via YAP1.

Primary hepatic malignant fibrous histiocytoma combined with invasion of inferior vena cava: A case report and literature review.

RATIONALE: Malignant fibrous histiocytoma (MFH), primary presented in liver, was very rare and displayed a poor prognosis because of high aggression. As a few of cases had been reported merely, we shared the case of primary hepatic MFH combined with invasion of inferior vena cava (IVC). PATIENTS CONCERNS: A 69-year-old women presented with abdominal pain. DIAGNOSES: Abdominal computed tomography and magnetic resonance imaging indicated a soft mass about 5.4 × 4.2 cm in the caudate lobe, accompanied with IVC invaded. INTERVENTIONS: After the multidisciplinary consultation, laparotomy was performed, followed by chemotherapy and radiotherapy. Primary hepatic MFH was demonstrated pathologically. Till now, the patient was alive for >22 months after surgery and no evidence of recurrence or distant metastasis was suspected. OUTCOMES: We discussed the integrated procedure of diagnosis and treatment, combined with data from literature review. LESSONS: To our knowledge, the primary hepatic MFH combined with invasion of IVC was hardly reported. Despite the poor prognosis, the comprehensive treatment integrating the surgery, chemotherapy, and radiotherapy showed the satisfactory disease-free and overall survival. However, further investigations are definitely warranted.

[Inhibition of malignant phenotype of HeLa cells by RNA interference of the telomerase activity].

OBJECTIVE: To investigate the biology of HeLa cells upon inhibition of human telomerase catalytic subunit (hTERT) gene by RNA interference in vitro. METHODS: Four shRNAs (A, B, C and D) targeting hTERT gene were designed and prepared by in-vitro transcription. The expression of hTERT gene was evaluated by immunofluorescent staining and telomeric repeat amplification protocol (TRAP) ELISA (TRAP-ELISA), after transient transfection of shRNAs by lipid formulation. Through the initial selection, shRNA (B) was noticed as the most efficient one in down-regulating hTERT gene and therefore was chosen as the ultimate shRNA used in the experimental groups. Those transfected by non-silencing RNAi were chosen as the control groups. Cell spreading and migration were studied by microscopy and cell adhesion to fibronectin (FN) was assayed by cell counting kit-8 (CCK-8). Cell invasion was assessed by Boyden chamber assay. RESULTS: Cell spreading study revealed that the rates of spreading cells in the experimental groups were (5.6 +/- 2.3)% at 30 min, and (26.3 +/- 6.1)% 2 h after the inoculation, respectively, whereas the rates of spreading cells in the control groups were (31.3 +/- 7.9)% and (79.4 +/- 4.8)%, respectively. There were significant differences between the two groups (P < 0.01). However, most of the cells in both groups became spreading after 24 h. Cell adhesion assay demonstrated that the rate of adhesion cells on FN in experimental groups was (67.2 +/- 2.8)%, less than that in control groups (83.7 +/- 5.4)% (P < 0.05). The relative migration distance was (27.1 +/- 6.2)% in the experimental group, lower than that of the control group (58.7 +/- 15.0)%. The invasion assay revealed that the invading cells were 75.7 +/- 14.5 in the experimental group, in contrast to 165.1 +/- 11.0 in the control group after 4 h incubation on matrigel. The difference between these two groups was significant (P < 0.05). CONCLUSION: In vitro shRNA silencing of hTERT gene can down-regulate the telomerase activity, leading to an inhibition of the malignant phenotype of HeLa cells, including decreased ability of cell spreading and adhesion, reduction of cell migration, and declined invasive ability through Matrige assay.

[The clinical characteristics of postinfectious irritable bowel syndrome in Qingdao].

OBJECTIVE: To investigate the clinical characteristics of postinfectious irritable bowel syndrome (PI-IBS) in Qingdao. METHODS: Two hundred and four PI-IBS and 2068 non-PI-IBS patients were investigated with questionnaire including general information, symptoms and quality of life scores with microecological study before and after therapy. RESULTS: (1) The morbidity rate of PI-IBS in female was 2. 2 times of that in male, which was similar to that in non-PI-IBS. (2) Brain work labors dominated in both PI-IBS and non-PI-IBS patients. (3) As to the simultaneous presence of extra-gastrointestinal symptoms, there was no statistical difference between the rate of physical symptoms in PI-IBS and non-PI-IBS patients (chi2 =10.5, P > 0.05), but the rate of mental symptoms was higher in PI-IBS than in non-PI-IBS patients, and the difference was significant (chi2 = 28.7, P < 0.05). (4) The alteration of intestinal microflora rate in PI-IBS was obviously higher than that in non-PI-IBS patients. (5) The quality of life scores in PI-IBS was improved after treatment with Birid Triple Viable , and there was significant difference (t = 3.8, P < 0.01), but there was no statistical difference in non-PI-IBS (t = 1.5, P > 0.05). CONCLUSION: There was some difference in certain clinical characteristics between PI-IBS and non-PI-IBS patients in Qingdao.

[Construction expression and biologic activity of recombinant human sCR1 eucaryotic cell].

AIM: To express human soluble complement receptor type 1(sCR1)protein using ferment cell secreting type carrier and study the extraorgan biologic activity of recombinant human sCR1 fusion protein. METHODS: Total human RNA was extracted from peripheral blood. The full length cDNA of human sCR1 gene was obtained by RT-PCR and them, cloned into Pichia pastoris eukaryotic expression vector pPIC9k to construct the recombinant plasmid pPIC9k-sCR1 containing human sCR1.After identified by DNA sequencing, the recombinant plasmid pPIC9k-sCR1 was transformed into Pichia pastoris SMD1168. The ferment cell line of the recombinant sCR1 which was chosen by G418 resistance was identified by PCR, After methanol induction, the expressed protein products were verified by SDS-PAGE and Western blot, purified by Ni(2+)-NTA agarose affinity chromatography, and its biologic activity was identified. RESULTS: The obtained Pichia pastoris secretion type yeast carrier pPIC9k-sCR1 was chosen by G418 and identified by PCR to get a highly copied and integral recombinant ferment cell line. The recombinant human sCR1 fusion protein was expressed by yeast cells containing pPIC9k-sCR1 induced by methanol. It was a protein band about M(r) 31 000 in gel, which could be identified by CD35 of anti-sCR1 protein monoclonal antibody with Western blotting technique. The highly purified sCR1 fusion protein and its biologic activity were detected obtained by Ni(2+)-NTA agarose affinity chromatography. CONCLUSION: The recombinant human sCR1 fusion protein can be highly expressed in the Pichia pastoris expression system, which resembles the human natural protein's antigenicity and biologic activity.