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Background: Hip fracture patients often have a large drop in hemoglobin (Hgb) concentration that is associated with the initial trauma. However, there is no study of a correlation between Hgb concentration at admission and short-term mortality. Thus, we evaluated a possible linear and nonlinear association between Hgb and mortalityfor older patients with hip fracture., Methods: Consecutive older patients who had hip fractures were screened between January 2015 and September 2019. Demographic and clinical characteristics were collected. Linear and nonlinear multivariate Cox regression models were used to identify association between Hgb at admission and mortality. All analyses were performed with EmpowerStats and R software., Results: Two thousand five hundred eighty-nine patients were included in the study. There were 849 men and 1740 women. The mean age was 79.6 ± 6.8 years. The mean follow-up was 39.0 months. Nine hundred seven (35.0%) patients died for all-cause reasons. The mean Hgb at admission was 11.07 ± 1.95 g/dL. Linear multivariate Cox regression models showed Hgb at admission was associated with mortality ([Hazard Ratio] HR 0.91, 95% CI 0.87-0.95, P < 0.0001) after adjusting for confounding factors. However, the linear association was unstable, and nonlinearity was found between Hgb at admission and mortality. The Hgb concentration of 9.8 g/dL was an inflection point. A Hgb at admission < 9.8 g/dL was associated with mortality (HR 0.81, 95% CI 0.74-0.89, P < 0.0001), whereas > 9.8 g/dL was not a risk factor for mortality (HR 0.98, 95% CI 0.92-1.04, P = 0.4730)., Conclusions: The Hgb concentration at admission was nonlinearly associated with mortality of older patients with hip fracture, and Hgb at admission < 9.8 g/dL was a risk predictor of 3-year mortality., Research Registration: ChiCTR2200057323., (© 2023. The Author(s), under exclusive licence to European Geriatric Medicine Society.)

Objective: To compare the clinical outcomes of staged total knee arthroplasty (TKA) performed on both knees in the same patient using gap balancing (GB) and measured resection (MR) techniques, respectively. Methods: The clinical data of 57 patients undergoing bilateral staged TKA at the Xi'an Jiaotong University Affiliated Honghui Hospital from July 2018 to January 2020 were analyzed. Using the random number table, MR or GB technique was selected when patients underwent primary TKA, and contralateral procedure was done with another technique. The procedures were performed by one chief surgeon, and the same prosthesis was chosen for all the procedures. The two osteotomy techniques for TKA were compared in terms of surgical status, radiographic data, functional recovery and satisfaction rate. Results: Total of 57 patients, including 16 males and 41 females, were included in the study with a mean age of (68.5±4.6) years (59-79 years) at primary TKA. All patients were followed up for (29.6±4.5) months (22-39 months). The interval between the two procedures was (4.7±3.0) months (0.5-12.0 months). Postoperative drainage was less in the GB side when compared with that in the MR side [(93.6±22.2) ml vs (109.9±36.9) ml, P =0.003]. At the 1-month postoperative follow-up, the visual analogue scale (VAS) of pain was lower on the GB side (3.0±0.8) than on the MR side (3.5±1.2), the range of motion (ROM) was higher on the GB side (105.7°±8.2° vs 100.2°±7.5°), the Knee Society Score (KSS) was higher on the GB side (78.5±5.4 vs 74.2±6.3), and the Western Ontario and McMaster University (WOMAC) score was lower on the GB side (35.4±5.5 vs 38.0±6.3), there were significant differences in the up-mentioned indexes between the two groups (all P <0.05). However, the repeated-measures analysis of variance indicated that there was no significant difference in VAS score, ROM, KSS score and WOMAC score between the two techniques (all P >0.05). The satisfactory rate of GB technique was 84.2%(48/57), ant it was 86.0%(49/57) with MR technique ( P =0.446). There was also no significant difference between the two techniques in terms of complications ( P =0.754). Conclusion: Both the GB and MR technique result in good knee function with similar clinical outcomes in patients receiving TKA in both knees for osteoarthritis without significant deformity.

Background: During total knee arthroplasty (TKA), surgeons mobilize the patella to facilitate clear visualization of the articular surfaces and allow better prosthesis placement. According to the manipulation, this manipulation can be divided into patellar eversion and noneversion. However, the effect of patellar eversion in TKA is controversial, with substantial variability in clinical practice. This systematic review is aimed at assessing the adverse effects of patellar eversion and patellar noneversion duration in TKA., Methods: This updated systematic literature review identified randomized controlled trials comparing patellar eversion and noneversion durations in TKA. Two investigators independently extracted data and evaluated the quality of the studies. A meta-analysis was performed using RevMan version 5.3., Results: Nine studies with a total of 608 patients (730 knees) were included. Of these, 374 knees were classified in the eversion group and 356 knees in the noneversion group. The quality of the studies was high. The results showed that patellar eversion could increase the postoperative complication rate (relative risk [RR] = 1.67; 95% confidence interval [CI], 1.09-2.54; P = 0.02) and postoperative pain before discharge (mean deviation [MD] = 0.19; 95% CI, 0.04-0.34; P = 0.01), compared to noneversion. Additionally, patellar eversion could prolong the time until the patient is able to raise the leg while straightened (MD = 0.42; 95% CI, 0.24-0.59; P < 0.00001) and increase the length of stay (MD = 0.65; 95% CI, 0.05-1.25; P = 0.03). However, patellar eversion did not influence postoperative pain at 1 year (MD = 0.02; 95% CI, -0.23-0.28; P = 0.85), operative time (MD = -2.66; 95% CI, -8.84-3.52; P = 0.40), recovery of quadriceps force throughout the follow-up period, and Insall-Salvati ratio (MD = -0.04; 95% CI, [-0.11-0.02]; P = 0.23)., Conclusions: The patellar eversion could increase the postoperative complication rate and postoperative pain. Current evidence supports the avoidance of patellar eversion in TKA. Further large-sample and long-term trials are required to validate these results., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2022 Jun Wang et al.)

Objectives: The lung injury is often secondary to severe trauma. In the model of crush syndrome, there may be secondary lung injury. We hypothesize that high-mobility group box 1 (HMGB1), released from muscle tissue, mediates the apoptosis of alveolar epithelial cells (AEC) via HMGB1/Receptor of advanced glycation end-products (RAGE)/c-Jun N-terminal kinase (JNK) pathway. The study aimed to investigate how HMGB1 mediated the apoptosis of AEC in the rat model., Methods: Seventy-five SD male rats were randomly divided into five groups: CS, CS + vehicle, CS + Ethyl pyruvate (EP), CS + FPS-ZM1 group, and CS + SP600125 groups. When the rats CS model were completed after 24 h, the rats were sacrificed. We collected the serum and the whole lung tissues. Inflammatory cytokines were measured in serum samples. Western blot and RT-qPCR were used to quantify the protein and mRNA. Lastly, apoptotic cells were detected by TUNEL. We used SPSS 25.0 for statistical analyses., Results: Nine rats died during the experiments. Dead rats were excluded from further analysis. Compared to the CS group, levels of HMGB1 and inflammatory cytokines in serum were downregulated in CS + EP, CS + FPS-ZM1, and CS + SP600125 groups. Western blot and RT-qPCR analysis revealed a significant downregulation of HMGB1, RAGE, and phosphorylated-JNK in CS + EP, CS + FPS-ZM1, and CS + SP600125 groups, compared with the CS groups, excluding total-JNK mRNA. Apoptosis of AEC was used TUNEL to assess. We found the TUNEL-positive cells were downregulated in CS + EP, CS + FPS-ZM1, and CS + SP600125 groups., Conclusion: The remote lung injury begins early after crush injuries. The HMGB1/RAGE/JNK signaling axis is an attractive target to abrogate the apoptosis of AEC after crush injuries., (© 2022. The Author(s).)

Objective: Adding vitamin E to highly cross-linked polyethylene liners is frequently performed in clinical practice, aiming at reducing liner wear, increasing liner survival, and delaying revision surgery. This study is aimed at evaluating the revision rate, total femoral head penetration, and postoperative clinical function of highly cross-linked polyethylene liners with and without vitamin E in total hip arthroplasty., Methods: We conducted a systematic literature search to identify the use of highly cross-linked vitamin E liners compared to other liners in patients who received total hip arthroplasty (THA) before April 2021. The study quality assessment and data collection were conducted by two independent reviewers. Studies were artificially grouped, and vitamin E-enhanced liners (VE-PE) were compared with vitamin E-free liners (non-VE-PE). Analyses were executed using Review Manager version 5.4.1., Results: From the preliminary screening of 568 studies, fourteen studies met the research criteria. Compared to non-VE-PE, using VE-PE reduced the all-cause revision rate (odds ratio = 0.54; 95% confidence interval (CI) 0.40, 0.73; P < 0.0001). The total femoral head penetration of the VE-PE was lower than that of the non-VE-PE (mean difference = -0.10; 95% CI -0.17, -0.03; P = 0.007). However, there was no difference in clinical function, including the Harris Hip Score and EuroQol Five-Dimension Questionnaire scores., Conclusion: Compared to the liners without vitamin E, the addition of vitamin E to liners could reduce the all-cause revision rate by approximately 46% in the short-term follow-up. In addition, even though addition of vitamin E could also slow down femoral head penetration, there is no contribution to clinical function., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2021 Qian-Yue Cheng et al.)

Purpose: The study is aimed at assessing the role of preoperative computerised tomography (CT) examination in the quality of reduction and outcomes in elderly patients with intertrochanteric fracture., Methods: The elderly patients with an intertrochanteric fracture who were treated with proximal femoral nail antirotation were included. The patients were divided into the CT group and the no-CT group according to the presence of preoperative CT examination. Patients' baseline characteristics, quality of reduction, and function were recorded at follow-up. Functional outcomes were evaluated using the Harris hip scores (HHS)., Results: Totally, the study included 182 patients with intertrochanteric fractures, with 85 in the CT group and 97 in the no-CT group, admitted between January 2018 and June 2019. There was no difference in the quality of reduction, HHS, the fracture healing, or postoperative complications between the CT group and the no-CT group. However, the CT group experienced the shorter mean operative time and blood transfusion, compared to the no-CT group., Conclusions: The preoperative CT examination seems to be excessive for elderly patients with an intertrochanteric fracture., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2021 Tao Ma et al.)

Objective: To explore the similarities and differences in clinical pathological features and gene rearrangement of lupus erythematosus profundus(LEP) and subcutaneous panniculitis-like T-cell lymphoma(SPTL)., Methods: We compared the clinical presentations, histopathology, immunophenotypical features and T-cell receptor (TCR) gene rearrangement findings of 9 cases of LEP and SPTL., Results: For clinical features, most patients of LEP occurred on head and face without systemic symptoms. LEP patients responded well to hydroxycholorquine treatment with good prognosis. Most patients of SPTL tended to lower extremities involvement and accompanied with systemic symptoms, the patients with disseminated lesions or hemophagocytic syndrome(HPS) showed poorer prognosis. For histopathology, LEP patients showed dense inflammatory infiltrate in the dermis consisting predominantly of lymphocytes with less numbers of plasma cells. However, the dermis was spared in SPTL, and rimming of adipocytes and erythrophagocytosis was observed in SPTL. Lymphocytes of LEP expressing CD4 + /CD8 + , as well as clusters of CD20 + . CD138-positive cells and scatter of CD123-positive cells were also observed in LEP. Tumor cells of SPTL were CD4 - /CD8 + , βF1 + , CD138 - and CD123 - . The expression of TIA-1 or GrB was more favor in SPTL. Monoclonal T-cell receptor-γ gene rearrangement was found in 89% of SPTL patients while negative for LEP., Conclusion: Base on different clinical and pathological features, it is easy to distinguish LEP from SPTL. However, a minority of lesions in LEP localize at subcutaneous tissue, which may turn to immunophenotypical and TCR gene rearrangement test for diagnosis., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Background: Hydroa vacciniforme-like lymphoproliferative disorder (HVLPD) is a rare Epstein-Barr virus (EBV)-associated lymphoma that mainly affects children., Objectives: To examine the similarities and differences in the clinical pathological features, EBV infection status, and gene rearrangements in adults and children patients with HVLPD., Methods: We compared the clinical manifestations, histopathology, immunophenotypical features, EBV infection status, and T-cell receptor gene rearrangements in the adult and children HVLPD groups., Results: Clinical manifestations differed between children and adults groups. The children were characterized by blisters and severe facial swelling, whereas the adults were characterized by mild facial swelling and papules. Mosquito bite was significantly related to morbidity in the children group. Histologically, the number of mast cells in the adult group was greater than in the children group (P < 0.05). There were no significant differences in EBV infection status or TCR-γ gene rearrangements between 2 groups., Conclusions: There were differences in clinical pathology and prognosis between the 2 groups. A higher mast cell count and T-cell phenotype might be associated with a poor prognosis.

Soil fertility properties described by four physical indices and seven chemical indices in the 10-year long-term experiment of wheat fields in Weibei Highland were used to investigate the cumulative effect of 11 soil indices under six long-term conservation tillage treatments. The principal component analysis was performed to comprehensively evaluate the cumulative effect of soil fertility under different tillage treatments, so as to provide scientific basis for soil sustainable utilization and management in Weibei Highland. The results showed that, compared with continuous conventional tillage (CT/CT), soil bulk density by no-tillage/subsoiling rotational tillage (NT/ST), subsoiling/conventional tillage rotational tillage (ST/CT) and conventional tillage/no-tillage rotational tillage (CT/NT) decreased by 6.6%, 5.9% and 6.6%, respectively, and no significant difference was found between continuous no-tillage (NT/NT) and CT/CT. NT/NT had the highest content of >0.25 mm mechanical stable aggregate, and no-tillage/subsoiling rotational tillage (NT/ST) had the highest content of >0.25 mm water stable aggregate. Compared with CT/CT, the mechanical stable aggregate contents under other five tillage treatments increased by 1.7%-10.1%, soil organic matter increased by 0.6%-11.2%, and the contents of nitrogen, phosphorus, and potassium contents also increased. By principal component analysis, 11 soil indices were divided into two principal components to comprehensively characterize soil quality. The contribution rate of the first principal component was 75.5%, where the major contributors were soil organic matter, water stable aggregate, total nitrogen, total phosphorus, total potassium, alkaline hydrolysis nitrogen, available phosphorus, available potassium, bulk density and soil porosity. The contribution rate of the second principal component was 13.2%, where the major contributor was mechanical stable aggregate. Together, those two principal components accounted for 88.6% of the original variables. The soil fertility performance was in the sequence of NT/ST>ST/CT>CT/NT>NT/NT>ST/ST>CT/CT after gradually accumulating the effects of long-term conservation tillage. NT/ST was the most appropriate tillage patterns for the wheat fields in Weibei Highland, due to the resulting balanced and high-quality soil environment.

Background: Lateral compartmental osteoarthritis (LCOA), a major complication after medial mobile-bearing unicompartmental knee arthroplasty (UKA), is highly associated with the increased stress of the lateral compartment. This study aimed to analyze the effects on the stress and load distribution of the lateral compartment induced by lower limb alignment and coronal inclination of the tibial component in UKA through a finite element analysis., Methods: Eight three-dimensional models were constructed based on a validated model for analyzing the biomechanical effects of implantation parameters on the lateral compartment after medial Oxford UKA: postoperative lower limb alignment of 3° valgus, neutral and 3° varus, and the inclination of tibial components placed in 4°, 2° valgus, square, and 2° and 4° varus. The contact stress of femoral and tibial cartilage and load distribution were calculated for all models., Results: In the 3° valgus lower limb alignment model, the contact stress of femoral (3.38 MPa) and tibial (3.50 MPa) cartilage as well as load percentage (45.78%) was highest compared to any other model, and was increased by 36.75%, 47.70%, and 27.63%, respectively when compared to 3° varus. In the condition of a neutral position, the outcome was comparable for the different tibial tray inclination models. The inclination did not greatly affect the lateral compartmental stress and load distribution., Conclusions: This study suggested that slightly varus (undercorrection) lower limb alignment might be a way to prevent LCOA in medial mobile-bearing UKA. However, the inclination (4° varus to 4° valgus) of the tibial component in the coronal plane would not be a risk factor for LCOA in neutral position.

Background: The lateral pillar of the femoral head is an important site for disease development such as osteonecrosis of the femoral head. The femoral head consists of medial, central, and lateral pillars. This study aimed to determine the biomechanical effects of early osteonecrosis in pillars of the femoral head via a finite element (FE) analysis., Methods: A three-dimensional FE model of the intact hip joint was constructed from the image data of a healthy control. Further, a set of six early osteonecrosis models was developed based on the three-pillar classification. The von Mises stress and surface displacements were calculated for all models., Results: The peak values of von Mises stress in the cortical and cancellous bones of normal model were 6.41 MPa and 0.49 MPa, respectively. In models with necrotic lesions in the cortical and cancellous bones, the von Mises stress and displacement of lateral pillar showed significant variability: the stress of cortical bone decreased from 6.41 MPa to 1.51 MPa (76.0% reduction), while cancellous bone showed an increase from 0.49 MPa to 1.28 MPa (159.0% increase); surface displacements of cortical and cancellous bones increased from 52.4 μm and 52.1 μm to 67.9 μm (29.5%) and 61.9 μm (18.8%), respectively. In addition, osteonecrosis affected not only pillars but also adjacent structures in terms of the von Mises stress and surface displacement levels., Conclusions: This study suggested that the early-stage necrosis in the femoral head could increase the risk of collapse, especially in lateral pillar. On the other hand, the cortical part of lateral pillar was found to be the main biomechanical support of femoral head.

MYBA2 transcription factor (Myb-related gene) affects the coloring in grapevine berry and plays an important role in the biosynthesis of anthocyanin. The MYBA2 gene was cloned from Vitis vinifera L. cv. Cabernet Sauvignon and polyclonal antibodies for VvmybA2 were prepared. The VvmybA2 gene expression patterns were observed in seven tissues (the leaf, stem, flower, bud, root, berry, and tendril) and during the berry development stage at transcriptional and translational levels, respectively. The results indicated that the expression of VvmybA2 was approximately 11-fold higher in the berry than that in the other six tissues, and increased rapidly from 60 days after full bloom reaching a maximum on day 80. Furthermore, both the anthocyanin content and UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT) gene expression levels increased rapidly 60 days after full bloom. Moreover, correlation analysis indicated that the transcriptional and translational expression levels of the VvmybA2 gene were significantly positively correlated with not only UFGT and DFR genes but also with the anthocyanin content during berry development. These results suggested that VvmybA2 could not only regulate the transcription of both UFGT and DFR but also is involved in the expression of the UFGT gene associated with color determination in grape berries.

The aim of this study was to investigate the mRNA expression levels of suppressor of cytokine signaling 1 (SOCS1) in patients with systemic lupus erythematosus (SLE) compared with healthy controls. The associations of systemic lupus erythematosus disease activity index scores and clinical features of SLE with the expression levels of SOCS1 mRNA were also evaluated. Real-time quantitative reverse transcription-polymerase chain reaction was applied to detect the mRNA expression levels of SOCS1 in peripheral blood mononuclear cells from 34 patients with SLE and 34 healthy controls. The mRNA expression level of SOCS1 was significantly decreased in SLE patients in comparison with healthy controls (Z = -4.207, P < 0.001). Lower SOCS1 mRNA expression was detected in active SLE patients when compared with inactive ones (Z = -2.428, P = 0.015). There was no significant difference found for the SOCS1 mRNA levels between SLE patients with nephritis and those without (Z = -0.642, P = 0.521). The presence of photosensitivity, proteinuria, positive antinuclear antibody, and C4 decline were associated with SOCS1 mRNA levels in SLE patients (all P < 0.05). Furthermore, the SOCS1 mRNA expression was negatively correlated with disease activity (r s  = -0.372, P = 0.030). Our results suggest that the dysregulation of SOCS1 might be associated with the pathogenesis of SLE.

To examine the effect of postharvest ultraviolet C (UV-C) irradiation on flavanol polyphenol accumulation in the grape berry, we investigated total flavanol polyphenol content, the enzyme activity of leucoanthocyanidin reductase (LAR), and transcription of Vv lar1 and Vv lar2 using spectrophotometry, real-time polymerase chain reaction, and western blot analysis in 5-year-old Vitis vinifera L. cv. Cabernet Sauvignon plants. Our results indicated that the accumulation of flavanol polyphenol reached its highest value when exposed to UV-C irradiation for 30 min. Additionally, UV-C irradiation induced the transcription of Vv lar1 and Vv lar2 and the synthesis of LAR1 and LAR2 proteins, resulting in increased accumulation of flavanol polyphenol in the grape berry. Moreover, these effects were associated with the length of time of UV-C irradiation.

Objective: To explore the correlation of the clone sources of MF-like cells in the epidermis and RS-like cells in the dermis of lymphomatoid papulosis (LyP), a compartive study of this two type cells in 8 LyP cases was carried out., Methods: 8 cases of LyP (4 type A cases and 4 type C cases) were detected the immunophenotype first, and then comparative study between the MF-like cells and RS-like cells was conducted on T-cell receptor (TCR) gamma-chain gene rearrangement by combinating laser capture microdissectinon (LCM) and PCR. PCR products of one patient which had positive bands were selected randomly for sequencing and sequence alignment analysis., Results: Monoclonal TCRgamma gene rearrangement was detected in 6 cases (75%) of the 8 cases. Detection rates of LyP type A and type C were 50% (2/4) and 100% (4/4), respectively. The rates of MF-like cells and RS-like cells were 75% (6/8) and 75% (6/8), respectively. Homology of the products with positive stripes in both MF-like cells and RS-like cells in one randomly selected case was compared and analyzed, and the results showed the homology of them was 106/108 (98%)., Conclusion: The MF-like cells in the epidermis and the RS-like cells in the dermis of LyP may be from the same clone.

Objective: Studies investigating the association between interleukin (IL)-4 gene promoter -590C/T (rs2243250) polymorphism and autoimmune diseases report conflicting results. To derive a more precise estimation of the relationship, a meta-analysis was performed., Methods: A systematic literature search was conducted to identify relevant studies. Pooled odds ratio (OR) with 95% confidence interval (CI) was used to estimate the strength of association., Results: A total of 6001 cases and 6788 controls from 24 studies were analysed. Significant association of the C allele of IL-4 rs2243250 polymorphism with rheumatoid arthritis (RA) was detected (odds ratio (OR) = 0.696, 95% confidence interval (CI) = 0.601-0.807). Stratification by ethnicity indicated an association between the IL-4 rs2243250 polymorphism and RA in Caucasians. Furthermore, the overall ORs of the associations between the C allele and multiple scleorosis (MS) were 1.340 (95% CI = 1.102-1.630). However, we failed to reveal any association between IL-4 rs2243250 polymorphism and systemic lupus erythematosus (SLE), type 1 diabetes (T1D) or Graves' disease (GD)., Conclusions: The present study suggests that the IL-4 rs2243250 polymorphism might be associated with genetic susceptibility to autoimmune diseases, including RA and MS., (© 2014 European Academy of Dermatology and Venereology.)

The results of studies on association between FCGR2A-R/H131 polymorphism and susceptibility to systemic lupus erythematosus (SLE) in the Asian population are controversial. To derive a more precise estimation on the genetic risk of FCGR2A-R/H131 variant for SLE in Asians, a meta-analysis was performed for genotypes R/R + R/H (dominant effect), R/R (recessive effect), and R allele in fixed/random effects models. Twenty studies involving 4,832 SLE patients and 6,190 controls were included. The meta-analysis showed that FCGR2A-R/H131 variant was associated with development of SLE in overall Asians both at allele or genotype level (R vs H, OR 1.201, 95 % CI 1.098-1.315; RR + RH vs HH, OR 1.369, 95 % CI 1.115-1.682; RR vs RH + HH, OR 1.305, 95 % CI 1.029-1.654). After stratification by ethnic, a significant association of R allele with susceptibility to SLE was observed in the Chinese population (R vs H, OR 1.104, 95 % CI 1.030-1.183). Evidence from subgroup analyses in non-Chinese populations (all Asians excluding Chinese population) also showed a significant association (R vs H, OR 1.354, 95 % CI 1.207-1.519; RR + RH vs HH, OR 1.705, 95 % CI 1.234-2.355). In addition, FCGR2A-R131 allele was associated with a 1.186-fold (95 % CI 1.043-1.349) greater risk for the occurrence of nephritis in the Asians population with SLE. After stratification by ethnic, this significant association was only consistently identified in non-Chinese populations (R vs H, OR 1.220, 95 % CI 1.002-1.486). In summary, FCGR2A-R/H131 polymorphism is associated with SLE and lupus nephritis in Asians.

The aim of this study was to estimate the associations between tumor necrosis factor (TNF) gene polymorphisms and type 1 diabetes (T1D) using meta-analysis. Relevant studies were searched using PubMed and Embase up to August 2013. A total of 32 comparisons from 21 studies examining the associations between TNF polymorphisms and T1D were included in the present meta-analysis. Our meta-analysis identified a significant association between TNF -308 A/G polymorphism A allele and T1D in all subjects [odds ratio (OR) 2.001, 95 % confidence interval (CI) 1.732-2.312). Significant associations of AA and AA+AG genotype of TNF -308 A/G polymorphism with genetic susceptibility to T1D were also found (OR 3.203, 95 % CI 2.373-4.324; OR 2.232, 95 % CI 1.881-2.649). After stratification by ethnicity, significant associations of T1D with TNF -308 A/G polymorphism under all genetic models (A allele and AA, AA+AG genotype) were still detected in European (OR 1.952, 95 % CI 1.675-2.274; OR 3.108, 95 % CI 2.169-4.455; OR 2.249, 95 % CI 1.870-2.706, respectively) and non-European populations (OR 2.152, 95 % CI 1.488-3.112; OR 3.439, 95 % CI 2.000-5.914; OR 2.207, 95 % CI 1.496-3.257, respectively). Our meta-analysis also revealed an association of TNF -857 T/C polymorphism T allele with T1D risk (OR 1.647, 95 % CI 1.431-1.896). Furthermore, analysis of TT and TT+TC genotype indicated the same result patterns as shown by the TNF -857 T/C polymorphism T allele (OR 2.206, 95 % CI 1.467-3.317; OR 1.762, 95 % CI 1.490-2.083). In conclusion, our meta-analysis results indicate that TNF -308 A/G and -857 T/C polymorphisms are involved in the genetic background of T1D.

The adventitious root of Tripterygium wilfordii was used as experiment material to study effects of various concentration of aspartic acid, isoleucine, cysteine and arginine in MS medium on the growth and triptolide, wilforgine, wilforine contents of the adventitious roots. The results showed that compared with the control, supplemented with 0.25 mmol x L(-1) aspartic acid at 3rd week, the growth of the adventitious roots only accounted for 80%, but the content of triptolide of the adventitious roots and the medium was 1.36, 1.30 times, the content of wilforgine was 1.16, 1.37 times, the content of wilforine was 1.22, 1.63 times, respectively. At 3rd week 0.05 mmol x L(-1) isoleucine, the growth of adventitious roots was 97.3%, wilforgine of adventitious roots and medium 1.02, 1.27 times, wilforine 1.36 times and 1.15 times. At 1st week 0.25 mmol x L(-1) cysteine, the growth of the adventitious roots comprised 77.5% of the control, while content of triptolide of adventitious roots reached 1.87 times. At 2nd week 1.00 mmol x L(-1) cysteine, the growth of adventitious roots was 44.6% of the control, the content of wilforine in medium was 2.97 times. At 3rd week 0.50 mmol x L(-1) arginine, the growth of adventitious roots was 124.2%, the content of wilforgine and wilforine was 1.3, 1.4 times, respectively.

Background: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by abnormal production of autoantibodies and proinflammatory cytokines. The clear pathogenesis of SLE has not been fully elucidated. Cytokine-mediated immunity has been showed to be involved in the pathogenesis of SLE., Objectives: The aim of this study was to investigate serum levels of cytokines (IL-19, IL-24, IL-26, IL-31, IL-32, IL-36) in SLE patients, in comparison with normal controls in a Chinese population., Materials and Methods: A total of 65 patients with SLE and 65 healthy volunteers were recruited for the current study. All serum levels of cytokines were measured by enzyme-linked immunosorbent assay (ELISA) kits., Results: Serum levels of IL-19, IL-24, IL-26, IL-31, IL-32 and IL-36 in SLE patients were not significantly different from the normal controls (all p > 0.05)., Conclusion: Serum levels of IL-19, IL-24, IL-26, IL-31, IL-32 and IL-36 in SLE patients were not markedly different from the normal controls. However, functional research should be discussed in future studies to elucidate the roles of these cytokines in SLE.

Emerging evidences were accumulated to support the view that aberrant interleukin-7 (IL-7) signaling might be associated with autoimmunity. Former studies demonstrated the single nucleotide polymorphism (SNP) rs6897932 C/T in the IL-7 receptor (IL-7R) gene was associated with susceptibility to autoimmune diseases, including multiple sclerosis and type I diabetes. Given these, this study was conducted to investigate whether an association existed between SNP rs6897932 and the susceptibility to systemic lupus erythematosus (SLE), a severe systemic autoimmune disease. In this context, 816 SLE patients and 816 controls from a Chinese population were recruited for this study, and the results showed that the major allele C of rs6897932 showed a higher frequency in SLE patients compared with controls (P = 0.039, C versus T); significant difference was also detected under a recessive model with regard to the distribution of genotype frequencies between SLE patients and controls (P = 0.041, CC versus CT + TT), which was not consistent with the results under a dominant model (P = 0.349, CC + CT versus TT). Moreover, association studies were also performed contraposing the relationship between the SNP rs6897932 C/T and lupus nephritis as well as 10 clinical features of SLE; however, no significant association signal was found regarding the distribution of allele and genotype frequencies between SLE patients positive and negative for the presence of 11 sub-phenotypes. In conclusion, the major allele C of SNP rs6897932 may be associated with increased SLE risk in Chinese populations, and further studies are still encouraged to shed light on the true associations between SLE and its susceptibility genes with respect to IL-7R gene.

Objective: The aim of this study was to determine whether CTLA-4 gene variants were associated with susceptibility to inflammatory bowel disease (IBD)., Methods: Meta-analysis was conducted on the association between CTLA-4 variants and IBD using: (1) allelic contrast, (2) the recessive model, and (3) the dominant model., Results: A total of 9 relevant studies including 1739 Crohn's disease (CD) cases, 10 relevant studies containing 1017 ulcerative colitis (UC) cases and 2685 healthy controls were involved in this meta-analysis. Overall, CTLA-4+49A/G, -318C/T and CT60 variants were not associated with IBD susceptibility in all genetic models (P>0.05). Stratification by ethnicity indicated a significant association between the CTLA-4+49A/G variant and CD in Caucasian group (GG vs. GA+AA: OR=0.723, 95% CI=0.564-0.926, P=0.010). In Asian group, meta-analysis showed a significant association between the CTLA-4 CT60 variant and UC (AA vs. AG+GG: OR=0.375, 95% CI=0.163-0.861, P=0.021)., Conclusions: Based on the published literature, this meta-analysis suggests that the CTLA-4+49A/G variant may be related to CD susceptibility in Caucasians, and the CTLA-4 CT60 variant may be associated with UC susceptibility in Asians., (Copyright © 2013 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)

Objective: The objective of this paper is to examine some solid tumors incidence in patients with systemic lupus erythematosus (SLE) derived from population-based cohort studies by means of meta-analysis., Methods: Relevant electronic databases were searched for studies characterizing the associated risk of overall malignancy and four site-specific malignancies (lung, liver, prostate, bladder cancer) in patients with SLE. The meta-analysis procedure was used to pool standardized incidence rates (SIRs) with 95% confidence intervals (CIs) to evaluate the association., Results: A total of seven cohort studies were identified, of which six provided the SIR for overall malignancy, seven reported the SIR for lung cancer, five for liver cancer, four for prostate cancer and six for bladder cancer. Overall, lung and liver cancers were more frequently observed in patients with SLE with SIR of 1.16 (95% CI = 1.12-1.21), 1.68 (95% CI = 1.33-2.13) and 2.44 (95% CI = 1.46-4.05), respectively. However, the risk of prostate cancer appeared to be somewhat reduced in male patients with SLE (SIR = 0.71, 95% CI = 0.57-0.89)., Conclusions: This meta-analysis shows that SLE patients are at increased risk of developing cancer, particularly of the lung, bladder and liver. However, males with SLE have a decreased risk of prostate cancer.

Objective: To determine whether the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) CT60 polymorphism (rs3087243) confers susceptibility to autoimmune thyroid disease (AITDs)., Methods: A meta-analysis was performed using: (1) allelic contrast, (2) recessive model and (3) dominant model. Electronic search of PubMed, Medline and Chinese National Knowledge Infrastructure (CNKI) was conducted to select studies., Results: Finally, a total of 20 separate studies were available for the current meta-analysis: Graves' disease (GD): 18 studies including 1 Iranian, 6 Caucasian and 11 Asian populations; Hashimoto's thyroiditis (HT): seven studies including one Iranian, three Caucasian and three Asian populations. A significant association was found between the CTLA-4 CT60 polymorphism (rs3087243) and GD, with regard to comparisons between allele and genotype frequencies (all p < 0.001). After stratification by ethnicity, significant relationships were consistently identified both in Caucasian and Asian populations. Furthermore, the association between this allelic variant and HT risk was also found in overall and Asian populations (OR: 1.26, 95% CI: 1.10-1.44; OR: 1.45, 95% CI: 1.19-1.76, respectively)., Conclusion: Taken together, our study suggested that the CT60 polymorphism (rs3087243) in CTLA-4 gene might confer susceptibility to the AITDs (GD/HT).

Interleukin-10 (IL-10) is a pleiotropic cytokine and plays an important role in the pathogenesis of systemic lupus erythematosus (SLE). Unlike IL-10 protein, few studies have focused on the potential association between IL-10 receptor (IL-10R) and SLE. The purpose of this study was to examine the association of a single nucleotide polymorphism, rs2834167 (A/G), in IL-10R2 gene (IL-10RB) with SLE in a Chinese Han population. A total of 667 patients with SLE and 676 healthy controls were enrolled in the present study. IL-10RB rs2834167 (A/G) polymorphism was specified from genomic DNA using TaqMan genotyping assay on a 7300 real-time reverse transcription polymerase chain reaction system. We found that the frequency of A allele for rs2834167 in patients (44.53 %) was significantly higher than in controls (40.16 %) (χ (2) = 5.24, P = 0.022). Allele A was associated with a 1.196-fold (95 % confidence interval (CI), 1.026-1.394) greater risk for the occurrence of SLE compared with the G allele. And both genotypes AG and AA were associated with the susceptibility to SLE as compared with the GG genotype (AG versus GG, odds ratio (OR) = 1.332; 95 %CI, 1.047-1.696; AA versus GG, OR = 1.373; 95 %CI, 1.004-1.878). We also found a statistical significance in the dominant model (AA + AG versus GG, OR = 1.343; 95 %CI, 1.070-1.687). However, no significant evidence for the association of IL-10RB rs2834167 (A/G) polymorphism with any clinical manifestations was detected. Our observations indicate that IL-10RB rs2834167 (A/G) polymorphism may be a potential biomarker for susceptibility to SLE.

Interleukin (IL)-7 is one of the IL-2 family cytokines comprised of IL-2, IL-4, IL-7, IL-9, IL-15, as well as IL-21. IL-7 is mainly secreted by stroma cells in primary lymphoid tissues, playing an essential role in the program of T cell development. Recently, studies have revealed that physiological function exerted by immunocytes can be influenced by aberrant IL-7 signaling, which is common in abnormal autoimmunity regulation. There is also increasing evidence that IL-7 is involved in several autoimmune diseases, such as rheumatoid arthritis, type I diabetes, multiple sclerosis and systemic lupus erythematosus, etc. Targeting components in IL-7 signaling pathways may have potential significance for treating numerous autoimmune diseases. In this review, we therefore summarize our current understandings regarding the relationship between IL-7 and autoimmune diseases so as to render more valuable information on this kind of research.

The aim of this study was to perform a meta-analysis of eligible studies to derive precise estimation of the association of interleukin-1 (IL-1), IL-10 and tumor necrosis factor (TNF)-α polymorphisms with Behcet's disease (BD). Odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of the association. A total of 4003 cases and 4748 controls in 19 eligible studies were included in the meta-analysis. We examined the relationship between seven single nucleotide polymorphisms (SNPs) in the above-mentioned three cytokine genes and susceptibility to BD. Meta-analysis indicated the association between the cytokine gene polymorphisms in all study subjects in the allelic model (TNF-α -308A/G: OR = 0.73, 95% CI: 0.61-0.88, P = 0.001; IL-10 -819C/T: OR = 0.72, 95% CI: 0.66-0.78, P < 0.001; IL-10 -592C/A: OR = 0.74, 95% CI: 0.64-0.86, P < 0.001); the dominant model (TNF-α -308A/G: OR = 0.77, 95% CI: 0.64-0.92, P = 0.004; IL-10 -1082G/A: OR = 1.64, 95% CI: 1.10-2.44, P = 0.014); the recessive model (TNF-α -308A/G: OR = 0.27, 95% CI: 0.12-0.65, P = 0.003; IL-10 -819C/T: OR = 0.71, 95% CI: 0.57-0.90, P = 0.004). However, no significant evidence for the associations of IL-1α -889C/T, IL-1β -551C/T, IL-1β -3962C/T polymorphisms with BD susceptibility was detected. The present study might suggest that TNF-α -308A/G, IL-10 -1082G/A, -819C/T, -592C/A polymorphisms are associated with BD susceptibility., (© 2013 Asia Pacific League of Associations for Rheumatology and Wiley Publishing Asia Pty Ltd.)

Background and Aims: Published data on the associations between interleukin-4 receptor (IL-4R) gene polymorphisms (Q551R, I50V) and rheumatoid arthritis (RA) risk are controversial. To quantitatively evaluate the relationships, a meta-analysis was performed., Methods: Studies were identified from the databases of PubMed, MEDLINE, Chinese Biomedical Literature Database, and Chinese National Knowledge Infrastructure, with the last report up to June 2012. The effect summary odds ratio (OR) and 95% confidence interval (CI) were obtained., Results: A total of six separate comparisons involving 2173 patients and 1892 controls were included to assess the association of IL-4R gene Q551R polymorphism and RA susceptibility. Overall, no significantly elevated RA risk was found in the meta-analysis. The pooled OR for the minor R allele was 0.942 (95% CI: 0.848-1.047, p=0.268) in patients with RA. After stratification by ethnicity, there was still no significant association detected in the European population (OR=0.979, 95% CI: 0.875-1.094). As for I50V polymorphism, there were four comparisons involving 1653 patients and 1584 controls in this meta-analysis. The pooled OR for the V allele was 1.104 (95% CI: 1.001-1.217) in RA, the V allele of the IL-4R gene I50V variant might be a risk factor for RA. However, the relationship between the V allele of IL-4R gene I50V polymorphism and rheumatoid factor positive in patients with RA was not identified through a minor meta-analysis, including four independent relevant comparisons., Conclusions: This meta-analysis indicates that the I50V polymorphism of IL-4R gene may confer susceptibility to RA; up to now, there is still not enough evidence to reveal the association of the IL-4R gene Q551R polymorphism with RA risk.

Purpose: Studies investigating the association between the tumor necrosis factor (TNF) gene polymorphisms and Behcet's disease (BD) report conflicting results. The aim of this meta-analysis was to assess the association between TNF gene polymorphisms and BD., Methods: A systematic literature search was conducted to identify all relevant studies. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were used to estimate the strength of the association., Results: A total of 16 articles, involving 1,708 patients with BD and 1,910 healthy controls, were included in the meta-analysis. Overall, a significant association was found between BD and the TNF -308A/G polymorphism (OR = 0.730, 95% CI = 0.608-0.877, p = 0.001). Meta-analysis of TNF -238A/G showed significant association with BD (OR = 1.512, 95% CI = 1.155-1.979, p = 0.003). The TNF -1031C allele showed significant association with BD (OR = 1.549, 95% CI = 1.190-2.015, p = 0.001). Similarly, the meta-analysis showed a significant association of the TNF -857T/C polymorphism with BD (OR = 0.758, 95% CI = 0.593-0.968, p = 0.027). Stratification by ethnicity revealed that the -308A/G and -857T/C polymorphisms were associated with BD in the Asian group, while the -238A/G and -1031C/T polymorphisms were associated with BD in the Caucasian population., Conclusions: The results of our meta-analysis suggest that TNF (-308A/G, -238A/G, -1031C/T, and -857T/C) polymorphisms are associated with susceptibility to BD.

To determine whether the tumor necrosis factor (TNF)-receptor associated factor 1/complement component 5 (TRAF1/C5) polymorphism (rs10818488) confers susceptibility to rheumatoid arthritis (RA) and systemic lupus erythematous (SLE), a meta-analysis was performed. A total of 11 studies with 17 comparisons (11 for RA, 6 for SLE) were available for this meta-analysis, which consisted of 13,456 patients, 12,259 controls for RA and 1,894 patients, 6,729 controls for SLE. A significant association of the A allele of TRAF1/C5 polymorphism (rs10818488) with RA susceptibility was detected in the North Africa population (OR=1.557, 95% CI: 1.225-1.977). Furthermore, the association between this allelic variant and SLE risk was additionally found in population of European (OR=1.247, 95% CI: 1.060-1.466). Analysis also showed the A/G allelic frequency of TRAF1/C5 variant (rs10818488), in different healthy populations, had a different distribution (χ(2)=269.41, P<0.001). Taken together, our study demonstrates that the TRAF1/C5 polymorphism (rs10818488) may confer susceptibility to RA in North Africa population, and in European population, it might be a contributory factor towards SLE., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Leprosy is a chronic granulomatous infection caused by Mycobacterium leprae, which primarily affects the skin and peripheral nerves. In this article, we present a 45-year-old man and a 39-year-old women who suffered from asymptomatic irregular erythemas on their trunk and extremities. Since both patients denied the history of exposure to leprosy patients and were absent clinical signs of superficial sensation dysfunction and enlarged peripheral nerves, they were diagnosed of mycosis fungoides and livedo reticularis clinically. Nevertheless the biopsies of erythemas showed perineural and periadnexal foamy-cell granulomas in the dermis and Fite staining revealed a large number of acid-fast bacilli. A diagnosis of multibacillary leprosy was made finally. These cases revealed that since leprosy is still epidemic in some remote area in China and in other developing countries and its clinical manifestations may be very weird sometimes, the dermatologists should be alert of it and skin biopsy could confirm the diagnosis.

Current research indicated that the resveratrol was mainly accumulated in the skin of grape berry, however, little is yet known about the distribution of resveratrol, as well as the regulation mechanism at protein level and the localization of stilbene synthase (malonyl-CoA:4-coumaroyl-CoA malonyltransferase; EC 2.3.1.95; STS), a key enzyme of resveratrol biosynthesis, in young grape plants (Vitis vinifera L. cv. Cabernet Sauvignon). Resveratrol, whose constitutive level ranged from 0.2 mg kg(-1) FW to 16.5 mg kg(-1) FW, could be detected in stem, axillary bud, shoot tip, petiole, root and leaf of grape plants. Among them, stem phloems presented the most abundant of resveratrol, and the leaves presented the lowest. Interestingly, the level of STS mRNA and protein were highest in grape leaves. And the analysis of immunohistochemical showed the tissue-specific distribution of STS in different organs, presenting the similar results compared with the amount of protein. And the subcellular localization revealed that the cell wall in different tissues processed the most golden particles representing STS. Subjecting to UV-C irradiation, resveratrol and STS were both intensely stimulated in grape leaves, with the similar response pattern. Results above indicated that distribution of resveratrol and STS in grape was organ-specific and tissue-specific. And the accumulation of resveratrol induced by UV-C was regulated by transcriptional and translational level of STS., (Copyright 2009 Elsevier Masson SAS. All rights reserved.)

Both phospholipase D (PLD, EC 3.1.4.4) and salicylic acid (SA) play important roles in response to external stimulation and activating defense system in plants. However, roles of the two signals in plants during the development of thermotolerance induced by low temperature acclimation remain unclear. In the experiment presented in the paper, grape berries (Vitis vinifera L. cv. Chardonnay) were pretreated at 8 degrees C for 3h and then transferred to 45 degrees C for heat stress. Compared with the control without low temperature pretreatment, membrane permeability and malondialdehyde (MDA) contents were reduced and the expression of HSP73 increased in the low temperature-pretreated berries under heat stress. During low temperature acclimation, PLD, SA and HSP73 could be activated. Meanwhile, the expression of HSP73 and the accumulation of free SA induced by low temperature can be inhibited by PLD activity inhibitor. All these results suggest that the activation of PLD is an early response to low temperature, and it is involved in the accumulation of free SA and the development of thermotolerance induced by low temperature acclimation.

To investigate whether phospholipase D (PLD, EC 3.1.4.4) plays a role in adaptive response of post-harvest fruit to environment, a PLD gene was firstly cloned from grape berry (Vitis Vinifera L. cv. Chardonnay) using RT-PCR and 3'- and 5'-RACE. The deduced amino acid sequence (809 residues) showed 84.7% identity with that of PLD from Ricinus communis. The secondary structures of this protein showed the characteristic C2 domain and two active sites of a phospholipid-metabolizing enzyme. The PLD activity and its expression in response to heat acclimation were then assayed. The results indicated PLD was significantly activated at enzyme activity, as well as accumulation of PLD mRNA and synthesis of new PLD protein during the early of heat acclimation, primary suggesting that the grape berry PLD may be involved in the heat response in post-harvest grape berry. This work offers an important basis for further investigating the mechanism of post-harvest fruit adaptation to environmental stresses.

The subcellular distribution and activity of glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) were studied in developing peach (Prunus persica L. Batsch cv. Zaoyu) fruit. Fruit tissues were separated by differential centrifugation at 15,000g into plastidic and cytosolic fractions. There was no serious loss of enzyme activity (or activation) during the preparation of fractions. G6PDH activity was found in both the plastidic and cytosolic compartments. Moreover, DTT had no effect on the plastidic G6PDH activities, that is, the redox regulatory mechanism did not play an important role in the peach fleshy tissue. Results from the immunogold electron-microscope localization revealed that G6PDH isoenzymes were mainly present in the cytosol, the secondary wall and plastids (chloroplasts and chromoplasts), but scarcely found in the starch granules or the cell wall. In addition to a decrease in fruit firmness, the G6PDH activity in the cytotolic and plastidic fractions increased, and anthocyanin started to accumulate during fruit maturation. These results suggest that G6PDH, by providing precursors for metabolic processes, might be associated with the red coloration that occurs in peach fruit.

Flavonoids are closely related to a plant's antioxidative ability. Because chalcone synthase (CHS) is the first enzyme to act as part of the flavonoid biosynthesis pathway, its expression and regulation are important. Here we present the expression of a full-length chs cDNA with 1225bp from grape seedlings as well as the preparation of an antibody against the expressed protein. A full-length chs cDNA was introduced into an expressed plasmid pET-30a(+) vector at the EcoRI and SalI restriction sites. pET-chs was found to be highly expressed in Escherichia coli BL21(DE3) pLysS cells with isopropyl-beta-d-thiogalactoside (IPTG) induction. A fusion protein with the His.tag label was purified by Ni-NTA His. Bind Resin and then used as the antigen to immunize a New Zealand rabbit. The resulting antiserum was then further precipitated by 50% saturated ammonium sulfate and DEAE-Sepharose FF column chromatography to obtain the immunoglobulin G (IgG) fraction. The resulting antibody was found capable of immuno-recognizing the CHS of the crude protein extracts from different grape tissues with a molecular mass of 43kDa.

The phenylpropanoid pathway yields a variety of phenolics that are closely associated with fruit qualities in addition to structural and defense-related functions. However, very little has been reported concerning its metabolism in fruit. This experiment was designed to assess changes of eleven phenolic acids in grape berry (Vitis vinifera L. cv. Cabernet Sauvignon) and explore both the activities and amounts of three key enzymes--phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate:coenzyme A ligase (4CL)--catalyzing the biosynthesis of these compounds during berry development. Finally, the subcellular localizations of the enzymes within berry tissues were also investigated using immuno-gold electron microscopic technique. The results indicated that the contents of gallic, protocatechuic, gentisic and caffeic acid all changed drastically during berry development, while other compounds containing p-hydroxybenzoic, vanillic, syringic, chlorogenic, p-coumaric, ferulic and sinapic acid varied only slightly. Activities of PAL, C4H and 4CL showed similar pattern changes with two accumulated peaks throughout berry development. In addition, their activities all showed a highly positive correlation with the total contents of phenolic acids, whereas the immunoblotting analysis showed that changes in enzyme activities were independent of the enzyme amounts. Results from the subcellular-localization study revealed that PAL was mainly present in the cell walls, secondarily thickened walls, and the parenchyma cells of the berry mesocarp cells, C4H was found primarily in the chloroplast (plastid) and nucleus and 4CL predominantly in the secondarily thickened walls and the parenchyma cells of mesocarp vascular tissue.