Sophoridine exerts anti-arthritic effects on fibroblast-like synoviocytes and collagen-induced arthritis in rats.

The discovery of alternative medicines with fewer adverse effects is urgently needed for rheumatoid arthritis (RA). Sophoridine (SR), the naturally occurring quinolizidine alkaloid isolated from the leguminous sophora species, has been demonstrated to possess a wide range of pharmacological activities. However, the effect of SR on RA remains unknown. In this study, the collagen-induced arthritis (CIA) rat model and tumor necrosis factor alpha (TNFα)-induced fibroblast-like synoviocytes (FLSs) were utilized to investigate the inhibitory effect of SR on RA. The anti-arthritic effect of SR was evaluated using the CIA rat model in vivo and TNFα-stimulated FLSs in vitro. Mechanistically, potential therapeutic targets and pathways of SR in RA were analyzed through drug target databases and disease databases, and validation was carried out through immunofluorescence, immunohistochemistry, and Western blot. The in vivo results revealed that SR treatment effectively ameliorated synovial inflammation and bone erosion in rats with CIA. The in vitro studies showed that SR could significantly suppress the proliferation and migration in TNFα-induced arthritic FLSs. Mechanistically, SR treatment efficiently inhibited the activation of MAPKs (JNK and p38) and NF-κB pathways in TNFα-induced arthritic FLSs. These findings were further substantiated by Immunohistochemistry results in the CIA rat. SR exerts an anti-arthritic effect in CIA rats through inhibition of the pathogenic characteristic of arthritic FLSs via suppressing NF-κB and MAPKs (JNK and p38) signaling pathways. SR may have a great potential for development as a novel therapeutic agent for RA treatment.

NtGCN2 confers cadmium tolerance in Nicotiana tabacum L. by regulating cadmium uptake, efflux, and subcellular distribution.

General control non-derepressible-2 (GCN2) is widely expressed in eukaryotes and responds to biotic and abiotic stressors. However, the precise function and mechanism of action of GCN2 in response to cadmium (Cd) stress in Nicotiana tabacum L. (tobacco) remains unclear. We investigated the role of NtGCN2 in Cd tolerance and explored the mechanism by which NtGCN2 responds to Cd stress in tobacco by exposing NtGCN2 transgenic tobacco lines to different concentrations of CdCl2. NtGCN2 was activated under 50 µmol·L-1 CdCl2 stress and enhanced the Cd tolerance and photosynthetic capacities of tobacco by increasing chlorophyll content and antioxidant capacity by upregulating NtSOD, NtPOD, and NtCAT expression and corresponding enzyme activities and decreasing malondialdehyde and O2·- contents. NtGCN2 enhanced the osmoregulatory capacity of tobacco by elevating proline (Pro) and soluble sugar contents and maintaining low levels of relative conductivity. Finally, NtGCN2 enhanced Cd tolerance in tobacco by reducing Cd uptake and translocation, promoting Cd efflux, and regulating Cd subcellular distribution. In conclusion, NtGCN2 improves the tolerance of tobacco to Cd through a series of mechanisms, namely, increasing antioxidant, photosynthetic, and osmoregulation capacities and regulating Cd uptake, translocation, efflux, and subcellular distribution. This study provides a scientific basis for further exploration of the role of NtGCN2 in plant responses to Cd stress and enhancement of the Cd stress signaling network in tobacco.

Neuronal CFL1 upregulation in head and neck squamous cell carcinoma enhances tumor-nerve crosstalk and promotes tumor growth.

Head and neck squamous cell carcinoma (HNSCC) is a prevalent cancer type, marked by a pronounced nerve density within the tumor microenvironment and a high rate of perineural invasion (PNI). Growing evidence suggests that the nervous system plays a vital role in HNSCC progression. Yet, the mechanisms governing cancer-nerve interactions remain largely elusive. Our research revealed that cofilin-1 (CFL1) is significantly overexpressed in HNSCC and correlates with both PNI and unfavorable prognosis. Utilizing multiplex fluorescent immunohistochemistry, we have localized CFL1 chiefly to the nerves adjacent to tumor sites. Significantly, it is the elevated expression of CFL1 in neuronal structures, rather than in the tumor cells, that aligns with diminished patient survival rates. We observed that HNSCC cells induced the expression of neuronal CFL1 and that the conditional knockout of neuronal CFL1 impedes tumor-nerve interactions. Both Gene Ontology functional enrichment analyses and Gene Set Enrichment Analysis demonstrate that CFL1 expression in HNSCC is associated with specific biological processes, including "RIBOSOME," "PROTEASOME," and "cadherin binding." In summary, HNSCC promotes the expression of CFL1 in nerves, which is essential for cancer-nerve interactions. The neuronal CFL1 is associated with PNI and may be a potential molecular prognostic marker of poor survival in HNSCC.

Inhibitory effects of norcantharidin on titanium particle-induced osteolysis, osteoclast activation and bone resorption via MAPK pathways.

Wear particles generated from the surface of implanted prostheses can lead to peri-implant osteolysis and subsequent aseptic loosening. In the inflammatory environment, extensive formation and activation of osteoclasts are considered the underlying cause of peri-implant osteolysis. Current medications targeting osteoclasts for the treatment of particle-induced bone resorption are not ideal due to significant side effects. Therefore, there is an urgent need to develop more effective drugs with fewer side effects. Norcantharidin (NCTD), a derivative of cantharidin extracted from blister beetles, is currently primarily used for the treatment of solid tumors in clinical settings. However, the potential role of NCTD in treating aseptic loosening of the prosthesis has not been reported. In this study, the in vitro results demonstrated that NCTD could effectively inhibit the formation of osteoclasts and bone resorption induced by the RANKL. Consistently, NCTD strongly inhibited RANKL-induced mRNA and protein levels of c-Fos and NFATc1, concomitant with reduced expression of osteoclast specific genes including TRAP, CTR and CTSK. The in vivo data showed that NCTD exerted significant protective actions against titanium particle-induced inflammation and subsequent osteolysis. The molecular mechanism investigation revealed that NCTD could suppress the activations of RANKL-induced MAPK (p38, ERK). Overall, these findings support the potential use of NCTD for the treatment of aseptic loosening following total joint arthroplasty.

Relationship of Frailty with Kidney Function in Adults More Than 60-Years-Old: Effect of Using Different Formulas to Estimate Glomerular Filtration Rate.

Objective: Determine the relationship of renal function with frailty using different formulas for estimated glomerular filtration rate (eGFR). Methods: Individuals who were 60-years-old or more (n=507) were recruited from August 2020 to June 2021, and the FRAIL scale was used to classify them as non-frail or frail. The three equations used to compute the eGFR were based on serum creatinine (eGFRcr), cystatin C (eGFRcys), or SCr+CysC (eGFRcr-cys). Renal function was classified using eGFR and defined as normal (≥90 mL/min/1.73m2), mild damage (59-89 mL/min/1.73m2), or moderate damage (≤60 mL/min/1.73m2). The relationship of frailty with renal function was analyzed. A subset of participants (n=358) was used to analyze changes in eGFR from 1 January 2012 to 31 December 2021 according to frailty and using the different eGFR equations. Results: There were significant differences between the eGFRcr-cys and eGFRcr values in the frail group (P<0.05), but not the non-frail group; however, the differences between the eGFRcr-cys and eGFRcys values were significant in the frail and non-frail groups (P<0.001). Based on each eGFR equation, the prevalence of frailty increased as eGFR decreased (P<0.001), but there was no significant relationship after adjusting for age or the age-adjusted Charlson co-morbidity index. There were temporal declines in eGFR in all three frailty groups (robust, pre-frail, and frail), especially in the frail group (2.226 mL/min/1.73m2 per year; P<0.001). Conclusion: For older individuals who are frail, the eGFRcr value may not provide accurate estimates of renal function. Frailty is associated with a rapid decline in kidney function.

Recent progress of direct thiocyanation reactions.

Thiocyanates are common in natural products, synthetic drugs and bioactive molecules. Many thiocyanate derivatives show excellent antibacterial, antiparasitic and anticancer activities. Thiocyanation can introduce SCN groups into parent molecules for constructing SCN-containing small organic molecules. Among them, the direct introduction method mainly includes nucleophilic reaction, electrophilic reaction and free radical reaction, which can simply and quickly introduce SCN groups at the target sites to construct thiocyanates, and has broad application prospects. In this review, we summarize the research progress of direct thiocyanation in recent years.

Peimine suppresses collagen-induced arthritis, activated fibroblast-like synoviocytes and TNFα-induced MAPK pathways.

BACKGROUND AND PURPOSE: Peimine (PM), a main isosterol alkaloid component isolated from the bulbs of traditional Chinese herb Fritillaria cirrhosa D. Don, has been demonstrated to exhibit multiple pharmacological properties, including anti-inflammation, anti-cancer and pain suppression. However, its effect on rheumatoid arthritis (RA) remains unknown. In the present study, we investigated the effect of PM on collagen-induced arthritis (CIA) rats in vivo and its inhibition on destructive behaviors of arthritic fibroblast-like synoviocytes (FLSs) in vitro. METHODS: Arthritis was induced in rats by chicken type II collagen. Arthritis score, radiological evaluation, and histopathological assessment were used to evaluate the therapeutic effects of PM on CIA rats. EdU assay, wound healing assay and real-time PCR were used to examine the inhibitory effect of PM on proliferation, migration, and over-expression of pro-inflammatory cytokines in TNFα-induced arthritic FLSs. TRAP staining and scanning electron microscopy were used to analyze the effect of PM on osteoclastogensis and bone resorption. Western blot was used to reveal PM's molecular mechanism of action on RA. RESULTS: PM significantly suppressed synovitis and bone destruction in CIA rats. In vitro experiments showed that PM treatment significantly inhibited TNFα-induced destructive behaviors of arthritic FLSs, including over-proliferation, migration and over-expression of pro-inflammatory cytokines. Additionally, RANKL-induced osteoclast formation and bone-resorpting function were also inhibited by PM. Further molecular mechanism studies revealed that PM treatment significantly suppressed TNFα-induced activations of MAPKs (ERK, JNK and p38) in arthritic FLSs. CONCLUSION: Our findings provide strong evidence that PM has the potential to be developed as a therapeutic agent for patients with RA.

Piperlongumine Inhibits Titanium Particles-Induced Osteolysis, Osteoclast Formation, and RANKL-Induced Signaling Pathways.

Wear particle-induced aseptic loosening is the most common complication of total joint arthroplasty (TJA). Excessive osteoclast formation and bone resorptive activation have been considered to be responsible for extensive bone destruction and prosthesis failure. Therefore, identification of anti-osteoclastogenesis agents is a potential therapy strategy for the treatment of aseptic loosening and other osteoclast-related osteolysis diseases. In the present study, we reported, for the first time, that piperlongumine (PL), a key alkaloid compound from Piper longum fruits, could significantly suppress the formation and activation of osteoclasts. Furthermore, PL effectively decreased the mRNA expressions of osteoclastic marker genes such as tartrate-resistant acid phosphatase (TRAP), calcitonin receptor (CTR), and cathepsin K (CTSK). In addition, PL suppressed the receptor activator of nuclear factor-κB ligand (RANKL)-induced activations of MAPKs (ERK, JNK and p38) and NF-κB, which down-regulated the protein expression of nuclear factor of activated T-cells cytoplasmic 1 (NFATc1). Using a titanium (Ti) particle-induced calvarial osteolysis model, we demonstrated that PL could ameliorate Ti particle-induced bone loss in vivo. These data provide strong evidence that PL has the potential to treat osteoclast-related diseases including periprosthetic osteolysis (PPO) and aseptic loosening.

Evaluation of flow field in the anterior segment during irrigation and aspiration in cataract surgery with three-dimensional computational simulation.

INTRODUCTION: To quantitatively analyze the flow field and fluid pressure contour in the anterior segment during the irrigation and aspiration (I/A) procedure by means of computational fluid dynamics (CFD) simulation. METHODS: Four different models were established combining the irrigation sleeve, aspiration cannula, anterior chamber, and capsule to simulate various scenarios during the coaxial I/A procedure. Commercial CFX software was applied to perform the steady-state simulation with the k-ε turbulence model. The inlet boundary condition was a balanced salt solution with a 100 cm height of pressure, and the mass flow rate boundary condition set at 40 cc/min was applied at the outlet. RESULTS: The four models included the I/A tip located in the middle of the pupillary plane, the capsule equator, and the middle of the posterior capsule. The inlet/outlet pressure at the tip of the I/A cannula was almost the same in the four models. Following ejection into the anterior chamber and capsule bag, the streamline speed rapidly decreased. The pressure in the anterior chamber and capsule bag was approximately 60-70 mmHg. The area on the cornea and capsule closer to the initial streamline of the inlet irrigation possessed larger pressure. The differential pressure on the cornea and capsule was greater when the irrigation orifice directly facing the wall. CONCLUSION: The pressure varies at different positions in the anterior segment during the I/A procedure. The study suggests that specific precautions are required when the tip moves closer to the wall and the irrigation orifice is directed toward the opposite wall.

Epiphyseal Arterial Network and Inferior Retinacular Artery Seem Critical to Femoral Head Perfusion in Adults With Femoral Neck Fractures.

BACKGROUND: A better understanding of the blood supply of the femoral head is essential to guide therapeutic strategies for patients with femoral neck fractures. However, because of the limitations of conventional techniques, the precise distribution and characteristics of intraosseous arteries of the femoral head are not well displayed. QUESTIONS/PURPOSES: To explore the characteristics and interconnections of the intraosseous vessel system between different areas of the femoral head and the possible blood supply compensatory mechanism after femoral neck fracture. METHODS: The three-dimensional (3-D) structures of the intraosseous blood supply in 30 uninjured normal human femoral heads were reconstructed using angiography methods and microCT scans. The data were imported in the AMIRA® and MIMICS® software programs to reconstruct and quantify the extra- and intraosseous arteries (diameter, length). In a separate experiment, we evaluated the residual blood supply of femoral heads in 27 patients with femoral neck fractures before surgery by analyzing digital subtraction angiography data; during the study period, this was performed on all patients in whom hip-preserving surgery was planned, rather than arthroplasty. The number of affected and unaffected subjects included in the three groups (superior, inferior, and anterior retinacular arteries) with different types of fractures (Garden Types I-IV) were recorded and analyzed (Fisher's exact test) to reflect the affected degrees of these three groups of retinacular arteries in patients after femoral neck fractures. RESULTS: The main results of our cadaver study were: (1) the main blood supply sources of the femoral head were connected by three main network structures as a whole, and the epiphyseal arterial network is the most widely distributed and the primary network structure in the femoral head; (2) the main stems of the epiphyseal arteries which were located on the periphery of the intraosseous vascular system have fewer anastomoses than the network located in the central region; (3) compared with the round ligament artery and anterior retinacular artery, the inferior retinacular artery has a relatively large caliber. Digital subtraction angiography of the 27 patients with hip fractures indicated that the inferior retinacular arterial system had a high likelihood of being unaffected after femoral neck fracture (100% [14 of 14] in nondisplaced fractures and 60% [six of 10] in Garden Type III fractures). CONCLUSIONS: The epiphyseal arterial network and inferior retinacular arterial system appear to be two important structures for maintaining the femoral head blood supply after femoral neck fracture. Increased efforts to protect these key structures during surgery, such as drilling and placing internal implants closer to the central region of the femoral head, might be helpful to reduce the effect of iatrogenic injury of the intraosseous vascular system. CLINICAL RELEVANCE: 3-D anatomic evidence of intraosseous arterial distribution of the femoral head and the high frequency with which the inferior retinacular arteries remained patent after femoral neck fracture lead us to consider the necessity of drilling and placing internal implants closer to the central region of the femoral head during surgery. Future controlled studies might evaluate this proposition.

Vascular anatomy and clinical application of the free proximal ulnar artery perforator flaps.

BACKGROUND: There is a dearth of detailed published work on the anatomy of ulnar artery perforators. The objective of this study was to fully document the vascular basis of the free proximal ulnar artery perforator flap and report its use in reconstruction of the hand. METHODS: (1) The ulnar artery perforators were studied in 25 fresh cadavers and 10 cast preparations. Cadavers were injected with lead oxide for 3-dimensional reconstruction. The origin, course, and distribution of the ulnar artery perforators were comprehensively documented. (2) Between August 2011 and January 2013, 29 free proximal ulnar artery perforator flaps were utilized for reconstruction of soft-tissue defects of the hand in 25 patients. Flap size varied from 3.5 × 2.0 cm to 24.0 × 4.0 cm, with a consistent thickness of approximately 3 mm. RESULTS: (1) There were 7 ± 2.0 ulnar artery perforators. The average external diameter was 0.6 ± 0.2 mm. Each perforator supplied an average area of 26 ± 7.0 cm(2). Extensive anastomoses were found between the ulnar artery perforators and multiple adjacent source arteries. (2) All flaps survived. The clinical results were satisfactory after 10.2 ± 5.3 months of follow-up. The flaps were considered cosmetically acceptable by both patients and doctors. CONCLUSIONS: The main advantage of the proximal ulnar artery perforator flap is that it is a thin flap that is ideal for upper extremity reconstruction, either as proximally or distally based local perforator flap or as a free flap. The donor site is excellent, and the vascular anatomy is very consistent.

Montelukast attenuates neuropathic pain through inhibiting p38 mitogen-activated protein kinase and nuclear factor-kappa B in a rat model of chronic constriction injury.

BACKGROUND: Cysteinyl leukotrienes and their receptors have been shown to be involved in the generation of neuropathic pain. We performed this study to determine the antagonistic effect of montelukast, a cysteinyl leukotrienes receptor antagonist, on neuropathic pain and its underlying mechanism. METHODS: Neuropathic pain was induced by chronic constriction injury (CCI) of the sciatic nerve in rats. After CCI, rats were repeatedly administered montelukast (0.5, 1.0, and 2.0 mg/kg intraperitoneal, once daily) for a period of 14 days. Mechanical withdrawal threshold and thermal withdrawal latency were assessed before surgery and on days 1, 3, 5, 7, and 14 after CCI. The levels of interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α in the spinal cord were determined by enzyme-linked immunosorbent assay. The phosphorylation of p38 mitogen-activated protein kinase (MAPK) and activation of nuclear factor-kappaB (NF-κB) were assessed by Western blot. The expression of astrocyte marker glial fibrillary acidic protein and microglia marker Iba-1 and the coexpression of p-p38MAPK and Iba-1 or NF-κB and Iba-1 were observed by immunofluorescent staining. RESULTS: The CCI group displayed significantly decreased mechanical withdrawal threshold and thermal withdrawal latency on days 1, 3, 5, 7 and 14 compared with sham groups (P <0.05, P < 0.0001), which were markedly increased by montelukast (P < 0.05, P < 0.01, P <0.0001). After administration with montelukast for 14 days, as biological markers of inflammation, the levels of IL-1ß (P < 0.0001), IL-6 (P = 0.001 for low dosage, P < 0.0001 for middle and high dosages), and TNF-α (P =0.002, 0.001, < 0.0001 for low, middle, and high dosage, respectively) in the spinal cord were lower than those in the CCI group. Western blot analysis demonstrated that montelukast reduced the elevated expression of p-p38 MAPK (P =0.006, 0.015, < 0.0001 for low, middle, and high dosage, respectively) and NF-κB (P < 0.0001) in the spinal cord induced by CCI. Immunofluorescent staining showed that montelukast could inhibit CCI-induced activation of microglia but not astrocytes in the spinal cord. In addition, montelukast (2.0 mg/kg) significantly decreased the number of p38MAPK and Iba-1 or NF-κBp65 and Iba-1 double-positive cells. CONCLUSIONS: These results suggest that montelukast could effectively attenuate neuropathic pain in CCI rats by inhibiting the activation of p38MAPK and NF-κB signaling pathways in spinal microglia.

[Clinical features and management of pyothorax due to postoperative cervical anastomotic leakage in esophageal cancer surgery].

OBJECTIVE: To investigate the clinical characteristics and managements of pyothorax due to postoperative cervical anastomotic leakage after esophageal cancer surgery. METHODS: From January 2006 to January 2013, 3342 patients with esophageal carcinoma underwent esophagectomy and cervical esophagogastric anastomosis. Of them, 19 patients developed pyothorax following cervical anastomotic leakage and their clinicopathological data were analyzed retrospectively. RESULTS: All the patients underwent a cervical anastomosis via a three-incisional approach (right cervicothoracic mid-abdominal incision, RT group, n=1094) or a two-incisional approach (left cervicothoracic incision, LT group, n=2248). The total number of cervical anastomotic leakage cases was 237, of which 152 cases were in LT group (6.8%), and 85 cases in RT group (7.8%), respectively (P=0.287). The incidence of pyothorax was 2.0% (n=3) in LT group, and 18.8% (n=16) in RT group, respectively (P<0.01). Fourteen cases develop pyothorax within 3 days after operation. The main symptoms were high fever, dyspnea and chest pain. All the pyothorax patients received conservative treatments, including thoracic closed drainage, nasogastric tube placement, jejunal stoma, nutritional support, antibiotics and symptomatic treatment. Sixteen cases were cured, while 3 cases were dead. CONCLUSIONS: The right thoracotomy approach predisposes the cervical anastomotic leakage-associated pyothorax. Sufficient drainage and sufficient nutritional support are critical to the treatment.

Human papillomavirus infection and esophageal squamous cell carcinoma: a case-control study.

BACKGROUND: The risk factors for esophageal squamous cell carcinoma (ESCC) in the high-incidence areas of China remain unclear. METHODS: A total of 300 patients with ESCC and 900 controls matched for age and sex were enrolled in Anyang (China), a high-risk area for ESCC in China. In tumor tissue of the cases and in esophageal biopsies of controls, the presence of human papillomavirus (HPV) DNA was assessed by an SPF1/GP6(+)-mediated PCR followed by sequencing. The presence of serum antibody against the HPV-16 E7 oncoprotein was assessed by use of the ELISA. ORs with 95% confidence intervals (CI) were calculated via unconditional logistic regression models. RESULTS: The presence of HPV in the esophagus (OR, 6.4; 95% CI, 4.4-9.2) was associated with increased risk of ESCC. Moreover, infection with "oncogenic" types of HPV (OR, 10.3; 95% CI, 6.3-16.8) was more strongly associated with ESCC than other types of HPV (OR, 2.4; 95% CI, 1.4-4.2). The presence of HPV-16 (OR, 12.8; 95% CI, 7.6-21.7) was particularly strongly associated with ESCC. In addition, a higher proportion of cases than controls had serum antibodies against HPV-16 E7 (OR, 6.1; 95% CI, 3.7-10.0). CONCLUSION AND IMPACT: This study provides the strongest epidemiologic evidence to date in support of the important role of HPV in the development of ESCC in high-incidence areas of China.

[Chest wall resection and reconstruction for thoracic tumor 
invading the chest wall: a report of 12 cases].

BACKGROUND AND OBJECTIVE: Tumor involving the chest wall is a common clinical event, and if there are no distant metastases, complete resection of tumor and involved chest wall can give excellent results. The aim of this study is to report experience with chest wall resection and reconstruction (CWRR) for 12 patients who suffered thoracic malignant tumor involving chest wall, including the artificial materials used for reconstruction, soft tissue coverage, and our multidisciplinary CWRR approach. METHODS: All characteristics of 12 cases of CWRR from Oct 2005 to Apr 2011 were reviewed, including preoperative treatment, surgical approach, resection range, reconstruction methods, the local and systematic complications and postoperative survival. RESULTS: All 12 of these patients underwent radical resection and bony chest wall resection, with resultant bony chest wall defects ranging from 25 cm² to 700 cm², soft tissue defects of 56 cm² to 400 cm². The bony chest wall was reconstructed using polypropylene mesh, and repair of the soft tissue was carried out using the shifting muscle flaps, myocutaneous flaps and omental flaps. There was only one significant complication in these 12 cases where 1 case suffered respiratory failure and needed mechanical ventilation but recovered one month later. All 12 patients have survived to the end point of follow up. CONCLUSIONS: Only thoracic surgery and reconstructive surgery work together can complete the complex CWRR which according the tumor discipline. Thoracic surgeons as the leader and reconstructive surgeons as the subsidiary and be familiar with reconstruction materials of bony chest wall and appropriate choice of soft tissue coverage is the key to achieve radical surgery and to ensure long-term survival.

[Gene expression profiling study of HPV-16 positive and HPV-negative esophageal squamous cell carcinoma].

OBJECTIVE: To study the difference in gene expression between human papillomavirus (HPV)16-positive and HPV-negative esophageal squamous cell carcinoma(ESCC) . METHODS: Eight HPV 16-positive and seven HPV-negative ESCC specimens were evaluated by PCR. The samples were then determined for gene expression profiling using Solexa Sequencing Chip followed by bioinformatics analysis. RESULTS: A total of 796 differentially expressed genes between HPV 16-positive and HPV-negative ESCC were observed. Among them, 366 were up-regulated while 430 were down-regulated. Functional classification and pathway analysis showed that the functions of these genes were mostly related to tumor morphology, immune, and inflammatory response, cellular growth and proliferation and cellular movement. Of these, factors related to immune and inflammation were the most representative. CONCLUSION: Differences in immunologic factors may be associated with HPV infection in esophageal cancer.

Detection of HPV DNA in esophageal cancer specimens from different regions and ethnic groups: a descriptive study.

BACKGROUND: HPV has been found repeatedly in esophageal carcinoma tissues. However, reported detection rates of HPV DNA in these tumors have varied markedly. Differences in detection methods, sample types, and geographic regions of sample origin have been suggested as potential causes of this discrepancy. METHODS: HPV L1 DNA and HPV genotypes were evaluated in 435 esophageal carcinoma specimens collected from four geographic regions with different ethnicities including Anyang in north China, Shantou in south China, Xinjiang in west China, and the United States. The HPV L1 fragment was detected using SPF1/GP6+ primers. HPV genotyping was performed using genotype specific PCR. RESULTS: Two hundred and forty four of 435 samples (56.1%) tested positive for HPV L1. Significant differences in detection rate were observed neither among the three areas of China nor between China and the US. HPV6, 16, 18, 26, 45, 56, 57, and 58 were identified in L1 positive samples. HPV16 and 57 were the most common types in all regions, followed by HPV26 and HPV18. CONCLUSIONS: HPV infection is common in esophageal carcinoma independent of region and ethnic group of origin. Findings in this study raise the possibility that HPV is involved in esophageal carcinogenesis. Further investigation with a larger sample size over broader geographic areas may be warranted.

Matrix metalloproteinases expression correlates with survival in patients with esophageal squamous cell carcinoma.

OBJECTIVES: The matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases capable of degrading the extracellular matrix and play important roles in malignancies. We evaluated the expression of four MMPs in esophageal squamous cell carcinoma (ESCC), and assessed the association between MMP expression and clinicopathologic characteristics and disease-free survival time. METHODS: We evaluated MMP1, MMP7, MMP9, and MMP13 expression in tissues from 208 patients with ESCC using immunohistochemistry (IHC), and correlated MMP expression to clinicopathologic characteristics and disease-free survival time. To confirm MMP9 expression at different levels, we simultaneously performed RT-PCR, Western blotting, and IHC on tissues from a separate cohort of 23 patients with ESCC. RESULTS: IHC analysis showed that 63.0%, 41.8%, 49.0%, and 32.2% of 208 ESCC samples were positive for MMP1, MMP7, MMP9, and MMP13, respectively. MMPs were strongly expressed in the cytoplasm of cancer cells, especially in the invasive margin, and weakly expressed in stromal cells. No immunostaining was detected in non-cancerous esophageal mucosa. MMP9 expression was positively associated with poor tumor cell differentiation (p= 0.001), vessel permeation (p= 0.027), and lymph node metastasis (p= 0.027). MMP9 expression was a negative, independent predictor of disease-free survival time (Hazard ratio, 1.470; 95% CI, 1.105 approximately 1.955; p= 0.008). The expression of MMP7 (median survival time: 23 months for MMP7 positive patients, >77 months for MMP7 negative patients; p= 0.001) and MMP13 (median survival time: 18 months for MMP13 positive patients, 39 months for MMP13 negative patients; p= 0.014) correlated negatively with disease-free survival in relatively early stage ESCC patients. Co-expression of MMP7, MMP9, and MMP13 in relatively early stage ESCC samples identified patients with a poor prognosis (13 months median survival time) compared to those lacking MMP7, MMP9, and MMP13 expression (58 months median survival time, p < 0.001). CONCLUSIONS: MMP9 expression is a negative, independent prognostic factor in ESCC and correlates with tumor cell differentiation, vessel permeation, and lymph node metastasis. MMP7, MMP9, and MMP13 may function in early stage ESCC, and their co-expression predicts poor outcome for relatively early stage ESCC patients.

LMP7/TAP2 gene polymorphisms and HPV infection in esophageal carcinoma patients from a high incidence area in China.

Esophageal carcinoma is characterized by a widely ranged incidence variation among the different geographic regions. Anyang is a county in Henan Province of North China with the highest prevalence of esophageal carcinoma. Human papillomavirus (HPV) infection has been linked to the etiology of esophageal cancer in this area. In this study, we investigated correlations of the polymorphisms at low molecular weight polypeptide (LMP) and transporters with antigen processing (TAP) genes, with the risk of esophageal carcinoma. DNA extracted from either tumor specimens or esophageal epithelial cells was used to test HPV infection. Peripheral blood lymphocyte DNA was used for LMP/TAP genotyping. Polymerase chain reaction was performed to analyze HPV infection and LMP/TAP gene polymorphisms. The combined effect of LMP/TAP gene polymorphisms and HPV infection on esophageal carcinoma was analyzed by using unconditional logistic regression models. The TAP2 codons 379 isoleucine carriers and LMP7 codons 145 lysine carriers were found to be more susceptible to esophageal carcinoma (OR = 2.74, 95% CI = 1.15-6.49, P = 0.023 for TAP2; OR = 2.19, 95% CI = 1.09-4.37, P = 0.027 for LMP7). Patients carrying homozygous LMP7/TAP2 haplotype C, which contained the glutamine at LMP7 codons 145 and the isoleucine at TAP2 codons 379, were more prone to develop esophageal carcinoma (OR = 2.96, 95% CI = 1.13-7.81, P = 0.027). An additive effect on the risk of esophageal carcinoma development was found among individuals carrying LMP7/TAP2 haplotype C and infected by HPV (OR = 4.33, 95% CI = 2.53-7.42, P < 0.0001). LMP7/TAP2 haplotype C may act as the risk factor in esophageal carcinoma development and it may influence the tumorigenesis in HPV infected individuals.

Expression of 11 HOX genes is deregulated in esophageal squamous cell carcinoma.

PURPOSE: HOX genes are vital for all aspects of mammalian growth and differentiation, and recent data have shown that their deregulated expression is related to carcinogenesis. To date, there has been no systemic study on expression of HOX genes in esophageal carcinoma. We investigated the expression pattern of 39 known HOX genes in cancerous and noncancerous tissue from 36 patients with esophageal squamous cell carcinoma to determine whether their expression is altered in esophageal cancer. EXPERIMENTAL DESIGN: Thirty-six patients with resectable esophageal squamous cell carcinoma (ESCC) were enrolled in this study. Specific primers were designed for each of 39 HOX genes, and reverse transcription-PCR was done in cancerous and noncancerous samples of these 36 patients. Furthermore, the expression of HOXA9 protein was subjected to Western blot analysis in all 36 paired tissue samples. RESULTS: Eight of 39 HOX genes were expressed in cancerous but not in noncancerous tissue. Five of 39 HOX genes were expressed both in cancerous and noncancerous tissue. Of the latter, expression of HOXA7, HOXA9, and HOXC6 was significantly higher in cancerous tissue (P < 0.05). The remaining 26 HOX genes were not detected in either types of tissue. HOXA9 protein was expressed in both kinds of tissue (cancer tissue versus noncancerous mucosa: 0.34 +/- 0.32 versus 0.24 +/- 0.27, P = 0.121). CONCLUSIONS: This is the first comprehensive survey of 39 HOX gene expression in ESCC and noncancerous mucosa. Five of the 39 HOX genes were expressed in both types of tissue indicating their possible role in maintaining normal structure and function of adult esophageal mucosa. Eleven of the 39 HOX genes were deregulated in cancer tissue. These genes possibly participate in the carcinogenesis of ESCC.