[Status of Heavy Metal in Organic Fertilizers in Main Tea Growing Regions of China].

To provide guidance for the safe use of organic fertilizers and improve soil quality and tea safety, it is necessary to conduct systematic analyses of the heavy metal content of organic fertilizers applied in the main tea producing areas of China. In this study, we analyzed the heavy metal contents in organic fertilizer samples collected from 2017 to 2019. The risks of collected organic fertilizers from different areas and sources were calculated. The results showed that the average concentrations of ω(As), ω(Hg), ω(Pb), ω(Cd), ω(Cr), ω(Cu), ω(Zn), and ω(Ni) in the collected organic fertilizers were 4.60, 0.22, 27.1, 0.78, 27.9, 58.3, 250.1, and 16.3 mg·kg-1, respectively. According to the assessment standard in NY/T 525- 2021, the over-limit rates of As, Hg, Pb, Cd, and Cr were 6.19%, 1.33%, 4.42%, 4.42%, and 1.33%, respectively. With respect to the area, the qualified rates were 100% in Shaanxi, Jiangsu, Anhui, Fujian, and Guangxi; 80%-90% in Shandong, Zhejiang, Hubei, Sichuan, Yunnan, and Guangdong; and only 54.5% in Jiangxi. The qualified rates of sources were 100% in rapeseed cake, soybean cake, and pig manure; 95.8% in sheep manure; 91.7% in cow manure; 90.7% in chicken manure; 87.2% in manure of other animals; 82.4% in the mixture of plant and animal sources; 65.2% in other plant sources; and 63.6% in other sources. According to the recommended application rate, the accumulation rate of heavy metals in soil with pig manure, cow manure, chicken manure, and sheep manure would be much higher than that with rapeseed cake and soybean cake. The average accumulation rate of organic fertilizer from animal sources was 7-30 times higher than that from plant sources. Therefore, it is recommended to use rapeseed cake or soybean cake fertilizer in tea plantation and to increase the supervision of heavy metal accumulation in soil and tea in those high-risk areas.

[Clinical results of arthroscopic anterior cruciate ligament reconstruction with short femoral tunnel].

OBJECTIVE: To evaluate the clinical outcome of arthroscopic reconstruction of anterior cruciate ligament (ACL) with short femoral tunnel. METHODS: From May 2013 to June 2017, 128 patients with anterior cruciate ligament reconstruction were performed with Transportal technique. Among them, 32 cases had short femoral tunnel were included, including 13 males and 19 females, aged 25.8 (17 to 43) years old, with a mean history of (4.5±1.1) months. The tibial tunnels were drilled in the middle of the footprint of the ACL, and femoral tunnels were drilled by transportal technique. Grafts were fixed with Endobutton at the femoral side and with interference screw at the tibial side. The changes of symptoms and signs were observed and the anterior tibial displacement was measured. The function of knee joint was evaluated by Lysholm score and Tegner score. RESULTS: All patients were followed up for over 2 years. At the latest follow-up, 30 patients were negative and 2 patients were positive in knee shift test; 28 patients were negative in Lachman sign, 4 patients were positive in degree I; 30 patients were negative in anterior drawer test, 1 patient was positive in degree I and 1 patient was positive in degree II. The anterior displacement of the tibia increased by (2.6±1.8) mm compared with the healthy side, which was significantly different from that before operation (t=19.77, P<0.05). Lysholm score of 82.2±6.1 was significantly higher than that before operation (t=17.33, P=0.001). According to Lysholm score evaluation, 15 cases got an excellent result, 10 were good, 7 were fair, and no bad results, with a significant difference compared with that before operation (z=-7.151, P<0.05). Tegner motor function score of (7.4±0.6) was significantly different from that before operation (t=9.11, P=0.000 5). After operation, the knee joint movement ability of the patients improved significantly. Twelve patients could participate in antagonistic sports and 15 patients could participate in non-antagonistic sports. Fifteen patients were very satisfied with the curative effect, 13 patients were satisfied with the curative effect. CONCLUSIONS: The incidence of short femoral tunnel in anterior cruciate ligament reconstruction with transportal technique is 25%. At present, the clinical effect of patients with short tunnel is acceptable. However, due to the lack of comparative study, the effect of short tunnel on the curative effect is still unclear.

Role of autophagy in the progression of osteoarthritis: The autophagy inhibitor, 3-methyladenine, aggravates the severity of experimental osteoarthritis.

Accumulating evidence suggests that autophagy is closely related to the pathogenesis of osteoarthritis (OA). The aim of this study was to determine the changes in autophagy during the progression of OA and to elucidate the specific role of autophagy in OA. For this purpose, a cellular model of OA was generated by stimulating SW1353 cells with interleukin (IL)-1ß and a rabbit model of OA was also established by an intra-articular injection of collagenase, followed by treatment with the autophagy specific inhibitor, 3-methyladenine (3-MA). Cell viability was analyzed by MTS assay, and the mRNA expression levels of matrix metalloproteinases (MMP)-13 and tissue inhibitor of metalloproteinase (TIMP)-1 were determined by RT-qPCR. Cartilage degeneration was examined under a light microscope, and autophagosome and chondrocyte degeneration was observed by transmission electron microscopy. The protein expression of Beclin-1 and light chain 3 (LC3)B was evaluated by western blot analysis and immunofluorescence staining. We found that the autophagy was enhanced during the early stages and was weakened during the late stages of experimental OA. The inhibition of autophagy by 3-MA significantly aggravated the degeneration of chondrocytes and cartilage in experimental OA. Our results thus determine the changes in autophagy during different stages of OA, as well as the role of impaired autophagy in the development of OA. Our data suggest that the regulation of autophagy may be a potential therapeutic strategy with which to attenuate OA.

(5R)-5-hydroxytriptolide (LLDT-8) prevents collagen-induced arthritis through OPG/RANK/RANKL signaling in a rat model of rheumatoid arthritis.

(5R)-5-hydroxytriptolide (LLDT-8) extracts from Tripterygium have anti-inflammatory, antineoplastic and immunity adjustment functions. The present study used a collagen-induced arthritis (CIA) model to evaluate whether LLDT-8 prevents collagen-induced arthritis, and investigated the signaling underlying this. Male Sprague-Dawley rats were induced to generate CIA, mimicking rheumatoid arthritis (RA). The presence of arthritis was determined using RA progression scores. The inflammatory cytokines interleukin (IL)-1ß, IL-6 and nuclear factor-κB were detected using enzyme-linked immunosorbent assay kits. Induced nitric oxide synthase (iNOS) and matrix metalloprotease (MMP)-13 protein expression were measured using western blot analysis. Lastly, reverse transcription-quantitative polymerase chain reaction was used to evaluate osteoprotegerin (OPG) and receptor activator of nuclear factor κB (RANK) gene expression. LLDT-8 improved RA progression scores and reduced the incidence and severity of CIA. Furthermore, LLDT-8 administration inhibited collagen-induced inflammation and iNOS protein expression in arthritic rats. The current data indicated that MMP-13 production was suppressed and OPG/RANKL expression was increased by LLDT-8 treatment in the arthritic rat. The present results suggest that LLDT-8 attenuates CIA through OPG/RANK/RANK ligand signaling in a rat model of RA.

Increased receptor activator of nuclear factor κß ligand/osteoprotegerin ratio exacerbates cartilage destruction in osteoarthritis in vitro.

Osteoarthritis (OA) is a degenerative joint disease characterized by progressive cartilage destruction, matrix degradation and bony changes. Subchondral bone alterations in osteoarthritis are associated with cartilage destruction. It has previously been demonstrated that osteoprotegerin (OPG) and receptor activator of nuclear factor κß ligand (RANKL) mediate this process. The RANKL/OPG ratio is altered in OA chondrocytes compared with normal chondrocytes. In the pathogenesis of OA, abnormal expression levels of matrix metalloproteinase-13 (MMP-13) are secreted by chondrocytes has a vital role in the progression of cartilage erosion. In the present study, the effect of various RANKL/OPG ratios on MMP-13 expression levels was investigated in interleukin-1ß-stimulated SW1353 human chondrosarcoma cells. Cell viability was assessed by MTT assay and MMP-13 mRNA and protein expression levels were analyzed by quantitative reverse-transcription-quantitative polymerase chain reaction, ELISA and western blot analyses, respectively. The results demonstrated that an increase in MMP-13 mRNA and protein expression levels was observed with increasing RANKL/OPG ratio. These findings suggest that this mechanism may be used as a novel therapeutic strategy against OA.

[Effects of Enhanced Ultraviolet B Irradiation on Photosynthetic and Antioxidant System of Sorghum Seedlings].

The UV-B radiation on the surface of our planet has been enhanced due to gradual thinning of ozone layer. The change of solar spectrum UV-B radiation will cause damage to all kinds of terrestrial plants at certain degree. In this paper, taking breeding sorghum (Sorghum bicolor (L.Moench))variety Longza No.5 as sample, 40 µW·cm-2 UV-B radiation treatment was conducted on sorghum seedlings at two-leaf and one-heart stage and different time courses; then after a 2 d recovering, photosynthetic parameters were measured with a photosynthetic apparatus; the activities of antioxidant enzymes were detected as well. Our results revealed that, as the dosages of UV-B increasing, leaf browning injury was aggravated, plants dwarfing and significantly were reduced fresh weight and dry weight were observed; anthocyanin content was significantly increased; chlorophyll and carotenoid content significantly were reduced and net photosynthetic rate and chlorophyll fluorescence parameters were decreased. Meanwhile, with the increase in UV-B dosages, stomatal conductance, intercellular CO2 concentration and transpiration rate showed "down - up - down" trend; the activities of SOD and GR presented "down - up" changes; activities of POD and CAT demonstrated "down - up - down", and APX, GPX showed an "up - down - up" pattern. It is worth to note that, under the four-dose treatment, a sharp decline in net photosynthesis in sorghum seedlings was observed at 6 h UV-B treatment (equals to 2.4 J·m-2), and an obvious turning point was also found for other photosynthetic parameters and activities of antioxidant enzymes at the same time point. In summary, the results indicated that the enhanced UV-B radiation directly accounted for the damages in photosynthesis system including photosynthetic pigment content, net photosynthetic rate and chlorophyll fluorescence parameters of sorghum; the antioxidant system showed different responses to UV-B radiation below or above 6 h treatment: ASA-GSH cycle was more sensitive to low-dose UV-B radiation, while high-dose UV-B radiation not only undermined the photosynthesis system, but also triggered plant enzymatic and non-enzymatic antioxidant systems, resulting in leaf browning and necrosis,biomass accumulation reduction, plant dwarfing and even death.

[Alleviation of salt stress during maize seed germination by presoaking with exogenous sugar].

The maize variety Kenyu 6 was used to study the effects of exogenous glucose (Glc) and sucrose (Suc) on salt tolerance of maize seeds at germination stage under 150 mmol · L(-1) NaCl treatment. Results showed that under salt stress condition, 0.5 mmol · L(-1) exogenous Glc and Suc presoaking could promote seed germination and early seedling growth. Compared with the salt treatment, Glc presoaking increased the shoot length, radicle length and corresponding dry mass up to 1.5, 1.3, 2.1 and 1.8 times, and those of the Suc presoaking treatment increased up to 1.7, 1.3. 2.7 and 1.9 times, respectively. Exogenous Glc and Suc presoaking resulted in decreased levels of thiobarbituric acid reactive substances (TBARS) and hydrogen peroxide (H2O2) content of maize shoot under salt stress, which were lowered by 24.9% and 20.6% respectively. Exogenous Glc and Suc presoaking could increase the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione peroxidase (GPX), glutathione reductase (GR) and induce glucose-6-phosphate dehydrogenase (G6PDH) activity of maize shoot under salt stress. Compared with the salt treatment. Glc presoaking increased the activity of SOD, APX, GPX, GR and G6PDH by 66.2%, 62.9%, 32.0%, 38.5% and 50.5%, and those of the Suc presoaking increased by 67.5%, 59.8%, 30.0%, 38.5% and 50.4%, respectively. Glc and Suc presoaking also significantly increased the contents of ascorbic acid (ASA) and glutathione (GSH), ASA/DHA and GSH/GSSG. The G6PDH activity was found closely related with the strong antioxidation capacity induced by exogenous sugars. In addition, Glc and Suc presoaking enhanced K+/Na+ in maize shoot by 1.3 and 1.4 times of water soaking salt treatment, respectively. These results indicated that exogenous Glc and Suc presoaking could improve antioxidation capacity of maize seeds and maintain the in vivo K+/Na+ ion balance to alleviate the inhibitory effect of salt stress on maize seed germination.

Polymorphisms and phenotypic analysis of cytochrome P450 2D6 in the Tibetan population.

The cytochrome P450 2D6 enzyme (CYP2D6) metabolizes about 25% of prescribed drugs in the endoplasmic reticulum, and genetic polymorphisms in CYP2D6 can greatly affect its activity and lead to differences among individuals in drug efficacy and adverse drug reactions. To investigate genetic polymorphisms in CYP2D6 among Tibetan Chinese, we directly sequenced the whole gene in 96 unrelated, healthy Tibetans from The Tibet Autonomous Region of China and screened for genetic variants in the promoter, exons, introns, and 3'UTR. We detected fifty-one genetic polymorphisms in CYP2D6, and 16 of them are novel. The allele frequencies of CYP2D6*1, *2, *5, *10, *41, and *49 were 0.25, 0.43, 0.02, 0.29, 0.02, and 0.01, respectively. The frequency of CYP2D6*10, a putative poor-metabolizer allele, was lower in our sample population compared with that in the Han Chinese population (p<0.001). In addition, haplotype analysis allowed 15 CYP2D6 haplotypes to be classified into three groups. In conclusion, our results provide basic information about CPY2D6 alleles in Tibetans and suggest that the enzymatic activities of CYP2D6 may differ among the diverse ethnic populations of China. Our results provide a basis for safer drug administration and better therapeutic treatment among Tibetans.

Synthesis and antihistamine evaluations of novel loratadine analogues.

A series of loratadine analogues containing hydroxyl group and chiral center were synthesized. The effect of the synthesized compounds on the histamine-induced contractions of guinea-pig ileum muscles was studied. In addition, the in vivo asthma-relieving effect of the analogues in the histamine induced asthmatic reaction in guinea-pigs was determined. Most of the compounds exhibited definite H(1) antihistamine activity. The S-enantiomers, compounds 2, 4 and 8, are more potent than the R-enantiomers, compounds 1, 3 and 7. Compound 6 was the most active one among the eight synthesized compounds.

[Polymorphism of fifteen short tandem repeat loci in Tibetan of Changdu area].

OBJECTIVE: To investigate the genetic polymorphism of 15 autosomal short tandem repeat (STR) loci, i.e. D5S818, FGA, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX and D18S51, in Tibetan population of Changdu area, and to analyze the genetic relationship between this population and other Asian populations. METHODS: The 15 loci were amplified simultaneously using a multiplex PCR typing system. Amplified STR fragments were analyzed with an ABI PRISM 3100 Avant DNA Analyzer. Arlequin software version 3.1 was used to obtain allele frequencies and polymorphism parameters. RESULTS: There were 135 STR alleles in the 15 STRs of Changdu Tibetan, with allele frequencies ranging from 0.0065 to 0.5455. The average heterozygosity was 0.7340, the discrimination power was higher than 0.8 except for TPOX and TH01. The accumulative discrimination power was more than 0.9999998, and the accumulative non-parental exclusion probability was 0.99999997. CONCLUSION: The 15 STR loci of Changdu Tibetan possess characteristics of high genetic diversity. They can be employed in minority genetics investigation, and individual and paternity test in forensic medicine.

Accumulation and distribution of arsenic and cadmium by tea plants.

It is important to research the rules about accumulation and distribution of arsenic and cadmium by tea plants, which will give us some scientific ideas about how to control the contents of arsenic and cadmium in tea. In this study, by field investigation and pot trial, we found that mobility of arsenic and cadmium in tea plants was low. Most arsenic and cadmium absorbed were fixed in feeding roots and only small amount was transported to the above-ground parts. Distribution of arsenic and cadmium, based on their concentrations of unit dry matter, in tea plants grown on un-contaminated soil was in the order: feeding roots>stems approximately main roots>old leaves>young leaves. When tea plants were grown on polluted soils simulated by adding salts of these two metals, feeding roots possibly acted as a buffer and defense, and arsenic and cadmium were transported less to the above-ground parts. The concentration of cadmium in soil significantly and negatively correlated with chlorophyll content, photosynthetic rate, transpiration rate and biomass production of tea plants.

Effect of liming and seasonal variation on lead concentration of tea plant (Camellia sinensis (L.) O. Kuntze).

Tea is a widely consumed beverage. However, recent studies revealed that there were an increasing number of cases of tea products exceeding the former maximum permissible concentration (MPC) in China for Pb (2 mg kg(-1)). Tea Pb contamination is an issue affecting trade and consumer confidence. Root uptake of Pb could contribute significantly to Pb accumulation in tea leaves due to the strong acidity of many tea garden soils. We conducted pot and field experiments to evaluate the effect of liming on Pb uptake by tea plants on two highly acidic soils (pH3.6). Additions of CaCO(3) significantly increased soil pH by up to 1 unit and decreased soil extractable Pb by up to 32%. Liming resulted in a decrease in the proportion of Pb in the exchangeable and carbonate-bound fractions, with a concurrent increase in the fractions bound to Fe/Mn oxides and residues. Liming significantly decreased Pb concentrations of fine roots, stems and new shoots of tea plants in the pot experiment. In the field experiments, the effect of liming was not significant during the first year following CaCO(3) application, but became significant during the second and third years and Pb concentration in the new shoots was decreased by approximately 20-50%, indicating that liming of acidic tea garden soils is an effective way to reduce Pb contamination of tea. The study also reveals a distinct seasonal variation, with Pb concentration in the new shoots following the order of spring>autumn>summer.

Scale and causes of lead contamination in Chinese tea.

We investigated the scale and causes of Pb contamination in Chinese tea. Lead concentrations in 1,225 tea samples collected nationally between 1999 and 2001 varied from <0.2 to 97.9 mg kg(-1) dry weight (DW), with 32% of the samples exceeding the national maximum permissible concentration (MPC) of 2.0 mg kg(-1) DW and a significant difference between tea types. There was an increasing trend in tea Pb concentration from 1989 to 2000. Proximity to highway and surface dust contamination were found to cause elevated Pb concentrations in tea leaves. Furthermore, Pb concentration in tea leaves correlated significantly and positively with soil extractable Pb, and negatively with soil pH, suggesting that root uptake of Pb from soils also contributed to Pb accumulation in tea. Potential contributions to human Pb intake from drinking tea were small at the median or national MPC Pb values, but considerable at the highest concentration found in the study.