Modification of Enteromorpha prolifera with dielectric barrier discharge plasma to enhance malachite green adsorption.

Dyes, a kind of visible chemical, have severe deleterious effects on human health and ecological environment. In this work, batch biosorption experiments were carried out under various experimental conditions such as pH value and agitation time to optimize the potentiality of Enteromorpha prolifera for the removal of malachite green (MG) dye from aqueous solution (70·7%). Then, the algal biomass was treated with a dielectric barrier discharge (DBD) in helium for 4 and 10 min to enhance MG removal efficiency (84·7 and 96·6%). In addition, Fourier-transform infrared spectroscopy in combination with scanning electron microscopy was employed to monitor the chemical and physical changes of algal cells treated by DBD. This study illustrates that DBD may serve as an effective tool to activate the functional groups on the cell wall surface for dye binding, and it even offers an alternative new technique to improve the adsorption properties of native biosorbents for the removal of toxic dyes from wastewater.

[Effects of the resin base and retention depth on fracture resistance of molars restored with nano-ceramic endocrowns].

OBJECTIVE: To compare the effects of resin base and different retention depth on the fracture resistance of mandibular molars restored with nano-ceramic endocrowns. METHODS: Forty mandibular molars selected and randomly divided into 5 groups: ① The control group which was consisted of intact teeth, ② the non-resin base group, ③ the 2 mm retention depth group, ④ the 3 mm retention depth group, ⑤ the 4 mm retention depth group, respectively. After tooth preparation, in vitro root canal therapy was conducted, which was followed by endocrown design, production and adhesive of groups ②-⑤. All the samples were under load (N) of the universal mechanical testing machine after embedding. The fracture pattern of each sample was observed under stereomicroscope. Then the microstructure of the fracture surface was observed by scanning electron microscopy. RESULTS: The fracture loads of each group were respectively: the control group fracture load was (3 069.34±939.50) N; experimental groups: fracture load of (2 438.04±774.40) N for the group without resin base; fracture load of (3 537.18±763.65) N for the group with 2 mm retention depth. The fracture load of the retention depth 3 mm group was (2 331.55±766.39) N; the fracture load of the retention depth 4 mm group was (2 786.98±709.24) N. There was statistical significance in the effect of resin base and different retention depth on the fracture loads of molars restored with nano-ceramic endocrown (P < 0.05). Repairable fractures in each group were as follows: control group 2/8, non-resin base group 1/8, retention depth of 2 mm group 1/8, retention depth of 3 mm group 2/8, and retention depth of 4 mm group 0/8. The effects of the retention depth and the presence of resin base on the fracture resistance of the resin nano-ceramic endocrowns were statistically significant (P < 0.05). Scanning electron microscopy showed more arrest lines and small twist hackles on the fracture surface of the restorations with resin base (retention depths of 2 mm, 3 mm, and 4 mm), with cracks extending towards the root. In addition to the characteristics above, more transverse cracks parallel to the occlusal surface, pointing outwards from the center of the pulp cavity retention, were also observed on the fracture surface of the non-resin base restorations. CONCLUSION: When molar teeth with nano-ceramic endocrowns are restored, resin base and the retention depth of 2 mm help the teeth to obtain optimal fracture strength.

[Role of hepatic surgery in colorectal cancer multiple liver metastasis].

Colorectal cancer liver metastasis can be categorized as initially resectable and initially unresectable liver metastasis. Patients with initially resectable colorectal cancer liver metastases may benefit from hepatic surgery significantly,while those with initially unresectable metastases also have an opportunity to be treated radically by liver surgery after conversion therapy,so as to have a prolonged survival time. It is crucial to choose the right time and right way of surgical intervention. The timing depends on determination of tumor resectability,controlling of pre-operative systemic therapy and evaluation of liver function after systemic treatment. The selection of right way contains the election between synchronous operation and staged operation, resection margin and using of technologies such as laparoscope and associating liver partition and portal vein ligation for staged hepatectomy. This paper aims to explore the optimal timing for operation and the approaches of surgical method based on the research progress worldwide for prolonging the survival time of patients with colorectal cancer multiple liver metastases.

[Distribution of the anterior corneal astigmatism in over 140 000 patients with age-related cataract].

Objective: To analyze the distribution characteristics of the anterior corneal astigmatism in 140 000 cataract patients from 18 hospitals in China. Methods: Retrospective study. A total of 143 889 patients (143 889 right eyes) over the age of 40 years with age-related catarac were admitted to 18 Aier eye hospitals in China from July 2015 to October 2018. The average values of the three measurements of the magnitude of anterior corneal astigmatism, the meridian of corneal astigmatism, anterior chamber depth, corneal refractive power, and axial length measured by IOLMaster 500 were obtained. The data acquisition method of each sub-center was to collect and analyze the electronic case data in accordance with the inclusion and exclusion criteria, and to provide them for the sponsor Wuhan Aier Eye Hospital. Non-normal distribution data are presented as M (P25, P75). Mann-Whitney test, Kruskal-Wallis test, Chi-square test were used to analyze the distribution differences of the magnitude of corneal astigmatism and the meridian of corneal astigmatism in gender, age, anterior chamber depth, corneal refractive power and axial length. Results: Among the 143 889 patients, 84 319 were females and 59 570 were males, the median age was 72 (65, 78) years old, the median corneal astigmatism was 0.84 (0.51, 1.33) D; the corneal astigmatism was ≥0.75 D in 80 895 patients (56.22%) and was ≥1.00 D in 57 304 patients (39.83%). The median corneal astigmatism was 0.87 (0.53, 1.37) D in women and 0.82 (0.50, 1.29) D in men; with statistical difference (U=-14.891; P<0.05). The proportion of with-the-rule (WTR) astigmatism was 33.26% (28 046/84 319) for women and 34.26% (20 408/59 570) for men; The proportion of against-the-rule (ATR) astigmatism was 49.08% (41 385/84 319) for women and 46.91% (27 945/59 570) for men, with statistical difference (χ²=70.913; P<0.05). With the increase of age, the magnitude of corneal astigmatism first decreased from 0.94 (0.57, 1.48) D to 0.75 (0.46, 1.18) D, and then increased to 1.19 (0.74, 1.79) D, with statistical difference (H=1 263.438; P<0.05), and the change was at 61 to 70 years old. With the increase of age, the proportion of WTR astigmatism decreased from 77.50% (396/511) to 12.50% (3/24), the proportion of ATR astigmatism increased from 11.15% (57/511) to 79.07% (34/43), and the proportion of oblique astigmatism changed little from 17.02% (16/94) to 19.92% (245/1 230), the distribution difference was significant (χ²=10 174.496; P<0.05). As the anterior chamber became shallow, the magnitude of corneal astigmatism significantly increased from 0.82 (0.51, 1.31) D to 1.05 (0.61, 1.56) D, and the proportion of ATR astigmatism increased from 47.32% (60 207/127 227) to 51.69% (184/356) (H=409.961, χ²=120.995, both P<0.05). With the corneal refractive power rising, the magnitude of corneal astigmatism increased from 0.80 (0.49, 1.33) D to 0.95 (0.58, 1.53) D, the proportion of ATR astigmatism decreased from 52.84% (4 963/9 392) to 39.97% (9 023/22 577); the difference was significant (H=808.562, χ²=752.147, both P<0.05). When the axial length was>25.00 mm, the magnitude of corneal astigmatism was highest [1.04 (0.62, 1.65) D], and the proportion of ATR astigmatism was also highest [49.00% (10 964/22 376)]; the difference was significant (H=2 071.198, χ²=131.130, all P<0.05). Conclusions: The meridian of corneal astigmatism in middle-aged and elderly cataract patients is mainly ATR astigmatism. With the increasing of age, the magnitude of corneal astigmatism decreases first and then increases. The turning point from the proportion of WTR astigmatism to the proportion of ATR astigmatism is 65 years old. The shallower the anterior chamber is, the more the magnitude of corneal astigmatism and the proportion of ATR astigmatism increase. When the axial length is>25.00 mm, both the magnitude of corneal astigmatism and the proportion of ATR astigmatism reach the peak. (Chin J Ophthalmol, 2021, 57: 56-62).

[Comparative analysis of the clinical application of two femtosecond laser-assisted cataract operation platforms].

Objective: To evaluate the operability and clinical application effects of femtosecond laser-assisted cataract surgery systems of LenSx and LenSAR. Methods: This was a randomized controlled study. A total of 86 patients (90 eyes) who underwent femtosecond laser-assisted cataract surgery in Wuhan Aier Eye Hospital from April 2018 to November 2018 were enrolled and divided into two groups randomly, including 44 patients (45 eyes) in the LenSx group and 42 patients (45 eyes) in the LenSAR group. During the operation, the following observation indexes were obtained. Operational indicators included the number of docking attempts, anterior capsulotomy time, nucleus pre-treatment time, total femtosecond laser emission time, and total vacuum suction duration. Clinical outcome indicators included changes in the patient's intraocular pressure during femtosecond laser surgery, the rate of subconjunctival hemorrhage, capsulotomy integrity (yes/no), roundness and centricity of the anterior capsule opening (yes/no), the rate of anterior capsule opening tear, and the rate of posterior capsule rupture. The t-test, rank-sum test or chi-square test were used for statistical analysis. Results: There were no significant differences between groups in the age and the lens density (both P>0.05). The number of docking attempts in the LenSx group was 1 (1 to 4) and in the LenSAR group was 1 (1 to 2); there was statistically significant difference (Z =-2.23, P<0.05). The difference in the anterior capsulotomy time between the two groups was statistically significant [13.00 (10.00 to 22.00) s compared with 3.00 (1.00 to 3.00) s, Z=-8.71, P<0.05]. The femtosecond laser pre-nucleation time and total femtosecond laser emission time of the LenSx group were (16.67±3.36) s and (30.49±3.53) s, and those of the LenSAR group were (12.38±4.36) s and (15.36±4.29) s, respectively; the differences between the two groups were statistically significant (t=-5.23, -18.26; both P<0.05). The total vacuum suction duration in the LenSx group was (97.23±19.96) s, shorter than that in the LenSAR group [(123.76±16.81) s] (t=6.82, P<0.05). The intraocular pressure after femtosecond laser surgery in both groups was higher than that before surgery. The increase of intraocular pressure in the LenSAR group was (5.64±5.42) mmHg (1 mmHg=0.133 kPa), higher than that in the LenSx group [(2.99±4.66) mmHg] (t=-2.49, P<0.05). The rate of subconjunctival hemorrhage in the LenSx group was 33.3% (15/45), while it was 8.9% (4/45) in the LenSAR group; the difference between the two groups was statistically significant (χ²=6.67, P<0.05). There were no significant differences between groups in capsulotomy integrity, roundness and centricity of the anterior capsule opening, the rate of anterior capsule opening tear, and the rate of posterior capsule rupture (all P>0.05). Conclusion: The docking process of the LenSAR system is convenient, and there is less subconjunctival hemorrhage; the total vacuum suction duration of LenSx is short, and the increase of intraocular pressure is low. (Chin J Ophthalmol, 2020, 56: 530-535).

[Survey on the quadrivalent influenza vaccine intention and related factors of health care workers in the Pearl River Delta region from 2015 to 2017].

Objective: To analyze the quadrivalent influenza vaccine intention of 718 health care workers (HCWs) in the Pearl River Delta region from 2015 to 2017. Method: In May 2018, 718 HCWs from the department related to the diagnosis and treatment of influenza in 17 hospitals (6 tertiary hospitals, 5 secondary hospitals and 6 primary hospitals) from Guangzhou, Jiangmen, Zhuhai and Dongguan were selected by using stratified sampling method. Questionnaire survey and face-to-face interview were used to collect the information of influenza vaccination, the intention of the quadrivalent influenza vaccine, the acceptance of free and required vaccination policies, and recommendations for increasing influenza vaccination intentions from 2015 to 2017. The multivariate logistic regression was used to analyze factors associated with the vaccination intention. Results: A total of 718 HCWs were surveyed and 147 of them were interviewed face to face. Among them, the vaccination rate of primary hospitals [17.39%(40/230)] was higher than that of other hospitals (χ(2)=15.80, P<0.05). If the vaccine could be free, 84.82% (609/718) of HCWs would like to be vaccinated. The multivariate logistic regression showed that the factors, HCWs who were aged ≥50 years (OR=3.44, 95%CI:1.43-8.28), worked in department of prevention and health care (OR=2.35, 95%CI:1.16-4.75), learned about the quadrivalent influenza vaccine (OR=2.94, 95%CI:2.08-4.18), knowed that HCWs are priority (OR=2.33, 95%CI:1.56-3.48), and had a history of trivalent influenza vaccination from 2015 to 2017 (OR=4.70, 95%CI:3.08-7.15), were associated with the vaccination intention. Conclusion: HCWs in the Pearl River Delta region had weak inclination of getting quadrivalent influenza vaccine. HCWs who were age (≥50 years old), worked in department of prevention and health care, learned about the quadrivalent influenza vaccine, knowed that HCWs are priority, and had a history of trivalent influenza vaccination from 2015 to 2017 were factors positively associated with the vaccination intention.

[Comparison of the color difference between teeth underwent cold light whitening and two kinds of shade guides].

OBJECTIVE: To investigate which shade guide, Vitapan Classical or Vita Bleachedguide 3DMaster, is better matched with the color of teeth in judging whitening effect, by comparing the color difference between shade tabs and corresponding teeth underwent cold light tooth whitening. METHODS: A total of 60 patients underwent Beyond cold light tooth whitening from May 2014 to April 2016. The patients were divided into two experimental groups according to the shade guide used. Vitapan Classical shade guide was used to judge whitening effect in one group, and Vita Bleachedguide 3DMaster shade guide was used in another. Shade matching was carried out before and after whitening in both the two groups, and the results were recorded by digital photographs. Shade matching procedures were carried out by two doctors independently. If they chose the same tab, it would be seen as the shade matching result; While if they chose different tabs, another doctor would be invited to make a decision. Photographs were taken in preset conditions: intraoral photos of the full dentition in the front, and the proportion of shooting was 1:3; aperture was F22; shutter speed was 1/200; intensity of flash was M/8; ISO value was 200. The photographs were analyzed by Photoshop software. Chromatic values were measured, and color difference values were calculated. Measuring of chromatic values was carried out by three doctors independently, and all the photos were measured twice by each doctor. Six measure results of each photo were recorded, and the maximum and the minimum were excluded, then the mean was seen as the final result. The color difference values were compared by independent-sample t test. Besides, changes of shade tabs after whitening in the two groups were recorded. RESULTS: Color difference value was 5.06±1.71 in Vitapan Classical group, and 3.39±1.36 in Vita Bleachedguide 3D-Master group. There was statistically significant difference between the two groups (t=4.68,P<0.001). Change of shade tabs was 3.63±1.75 in Vitapan Classical group, and 2.23±1.01 in Vita Bleachedguide 3DMaster group. CONCLUSION: Vita Bleachedguide 3D-Master is better matched with the color of teeth, so it is preferred in judging the effect of cold light tooth whitening.

Temporal and spatial expression profiles of Frizzled 3 in the ovary during the estrous cycle.

Frizzled 3 is an important receptor in the Wnt/ß-catenin pathway, a conserved signaling pathway that regulates gene expression and controls diverse developmental processes. However, the role of this protein during follicular development in the adult ovary is not known. The present study was designed to investigate the expression and localization of Frizzled 3 mRNA and protein during the estrous cycle in the mouse ovary through in situ hybridization (ISH), real-time quantitative polymerase chain reaction, immunohistochemistry and western blot. ISH results showed that in proestrus, high expression of Frizzled 3 was found in the granulosa and stroma with weak levels in the corpus luteum. In estrus and diestrus, the stroma had high Frizzled 3 expression, but levels were low in granulosa cells and corpus luteum. In the metestrus, moderate expression of Frizzled 3 was found in the stroma but low to no expression was found in luteal cells and follicles. The mRNA and protein levels of Frizzled 3 were found to be the highest in proestrus and diestrus compared to estrus and metestrus (P < 0.05), confirming the ISH results. During estrus and diestrus, high Frizzled 3 expression was observed in the stroma and moderate levels in granulosa cells, and during estrus and proestrus, low expression was seen in the oocyte cell membrane. The western blot results further confirmed this change during the estrous cycle. Together, these results indicate that Frizzled 3 is involved in regulating follicular development and oocyte maturation during the estrous cycle.

Effects of dietary supplementation of synbiotics on growth performance, intestinal morphology, sIgA content and antioxidant capacities of broilers.

Today, several strategies are being used to decrease the serious effects of antibiotics abuse on broilers industry and public health, among which synbiotics are one of the most promising antibiotic alternative. This study was undertaken to assess the effects of synbiotics, which composed of probiotics (Bacillus subtilis) and prebiotics (xylooligosaccharide and mannanoligosaccharide), on growth performance, intestinal morphology, sIgA content and antioxidant parameters of broilers. Four hundred and fifty one-day-old commercial Cobb48 broilers were assigned to five treatments consisting of six replicates of 15 birds each pen. Five dietary treatments include basal diets (control), basal diets plus antibiotics (4 mg/kg Xanthomycin), basal diets plus 1 g of probiotics B. subtilis product/kg of diets (4 × 108  cfu/kg), basal diets plus 150 mg/kg xylooligosaccharide (35%) and 1 g/kg mannanoligosaccharide (75%), and basal diets plus synbiotics (1 g of probiotics B. subtilis product/kg of diets (4 × 108  cfu/kg), 150 mg/kg xylooligosaccharide (35%) and 1 g/kg mannanoligosaccharide (75%). The results demonstrated that on 21 and 42 days, dietary supplementation of the synbiotics significantly increased daily weight gain (p < 0.05), feed efficiency (p < 0.05), the villus height and villus:crypt ratio in the duodenum, jejunum and ileum (p < 0.05), as well as intestinal mucosa sIgA content (p < 0.05), serum T-SOD activity (p < 0.05) and lysozyme content (p < 0.05), comparing with control group. In conclusion, synbiotics (B. subtilis and xylooligosaccharide and mannanoligosaccharide) is one of the safe and ideal dietary supplementations to increase broilers' growth performance by improving small intestinal morphology, sIgA content and antioxidant capabilities.

The impact of plasma Epstein-Barr virus DNA and fibrinogen on nasopharyngeal carcinoma prognosis: an observational study.

BACKGROUND: The impact of combining plasma fibrinogen levels with Epstein-Barr Virus DNA (EBV DNA) levels on the prognosis for patients with nasopharyngeal carcinoma (NPC) was evaluated. METHODS: In this observational study, 2563 patients with non-metastatic NPC were evaluated for the effects of circulating plasma fibrinogen and EBV DNA levels on disease-free survival (DFS), distant metastasis-free survival (DMFS), and overall survival (OS). RESULTS: Compared with the bottom biomarker tertiles, TNM stage-adjusted hazard ratios (HR, 95% confidence intervals (CIs)) for predicting DFS in fibrinogen tertiles 2 to 3 were 1.26 (1.00 to 1.60) and 1.81 (1.45 to 2.26), respectively; HR for EBV DNA tertiles 2 to 3 were 1.49 (1.12 to 1.98) and 4.24 (3.27 to 5.49), respectively. After additional adjustment for established risk factors, both biomarkers were still associated (P for trend <0.001) with reduced DFS (HR: 1.79, 95% CI, 1.43 to 2.25 for top fibrinogen tertiles; HR: 4.04, 95% CI: 3.10 to 5.27 for top EBV DNA tertiles compared with the bottom tertiles). For patients with advanced-stage disease, those with high fibrinogen levels (3.34 g l(-1)) presented with worse DFS, regardless of EBV DNA 4000 or <4000 copies ml(-1) subgroup. Similar findings were observed for DMFS and OS. CONCLUSIONS: Circulating fibrinogen and EBV DNA significantly correlate with NPC patients survival. Combined fibrinogen and EBV DNA data lead to improved prognostic prediction in advanced-stage disease.

Alternative splicing, promoter methylation, and functional SNPs of sperm flagella 2 gene in testis and mature spermatozoa of Holstein bulls.

The sperm flagella 2 (SPEF2) gene is essential for development of normal sperm tail and male fertility. In this study, we characterized first the splice variants, promoter and its methylation, and functional single-nucleotide polymorphisms (SNPs) of the SPEF2 gene in newborn and adult Holstein bulls. Four splice variants were identified in the testes, epididymis, sperm, heart, spleen, lungs, kidneys, and liver tissues through RT-PCR, clone sequencing, and western blot analysis. Immunohistochemistry revealed that the SPEF2 was specifically expressed in the primary spermatocytes, elongated spermatids, and round spermatids in the testes and epididymis. SPEF2-SV1 was differentially expressed in the sperms of high-performance and low-performance adult bulls; SPEF2-SV2 presents the highest expression in testis and epididymis; SPEF2-SV3 was only detected in testis and epididymis. An SNP (c.2851G>T) in exon 20 of SPEF2, located within a putative exonic splice enhancer, potentially produced SPEF2-SV3 and was involved in semen deformity rate and post-thaw cryopreserved sperm motility. The luciferase reporter and bisulfite sequencing analysis suggested that the methylation pattern of the core promoter did not significantly differ between the full-sib bulls that presented hypomethylation in the ejaculated semen and testis. This finding indicates that sperm quality is unrelated to SPEF2 methylation pattern. Our data suggest that alternative splicing, rather than methylation, is involved in the regulation of SPEF2 expression in the testes and sperm and is one of the determinants of sperm motility during bull spermatogenesis. The exonic SNP (c.2851G>T) produces aberrant splice variants, which can be used as a candidate marker for semen traits selection breeding of Holstein bulls.

Effect of BMPRIB gene silencing by siRNA on apoptosis and steroidogenesis of porcine granulosa cells.

Bone morphogenetic proteins (BMPs) are the key factors in maintaining cell growth and differentiation in ovaries. BMPs initiate signaling by assembling BMP receptors and activating Smads, which in turn alter the expression of target genes. However, little is known about the effect of the deletion of the Bone morphogenetic protein receptor type IB (BMPRIB) on porcine granulosa cell (GCs). The objective of this study was to determine the effects of BMPRIB gene silencing, by small interfering RNA (siRNA), on the apoptosis and steroidogenesis of porcine GCs, and the expression of cell cycle-related and apoptosis-related genes. Results indicate that the BMPRIB siRNA caused specific inhibition of BMPRIB mRNA expression after transfection. Knockdown of the BMPRIB gene significantly inhibited porcine GCs proliferation and estradiol production, while inducing apoptosis of porcine GCs. Additionally, the declined expression of the BMPRIB gene changed the expressions of CylinD2, Cdk2, Bcl-2, and Cyp19a1. These findings provide an important role of BMPRIB in the regulation of apoptosis and steroidogenesis of porcine GCs.

[Current application and limitations of augmented reality in the stomatology].

Computer-assisted technology are gradually integrated into dental education and clinical treatment. As a cutting-edge technology in computer-aided medicine, augmented reality can not only be used as an aid to dental education by presenting three-dimensional scenes for teaching demonstration and experimental skills training, but also can superimpose virtual image information of patients onto real lesion areas for real-time feedback and intraoperative navigation. This review explores the current applications and limitations of augmented reality in dentistry to provide a reference for future research.

[Clinical effects of retrograde anterolateral thigh flaps in repairing anterior knee joint wounds under the concept of precise flap surgery].

Objective: To introduce the methods of retrograde anterolateral thigh flaps in repairing anterior knee joint wounds under the concept of precise flap surgery and to explore the clinical effects. Methods: A retrospective observational study was conducted. From August 2014 to March 2022, 7 patients with anterior knee joint wounds were treated with retrograde anterolateral thigh flap under the guidance of the concept of precise flap surgery in the 920th Hospital of Joint Logistic Support Force of PLA. Among them, 6 were males and 1 was female, aged 36 to 66 years. The sizes of wounds were 7 cm×5 cm to 15 cm×11 cm after debridement. All the patients were performed with computed tomography angiography (CTA), the donor and recipient sites were evaluated according to the precise flap surgery method, and the optimal pedicle, perforator, and pivot of flaps were chosen. The flap sizes were 10 cm×6 cm to 20 cm×9 cm, and all the donor sites of flaps were sutured directly. The consistency of the intraoperative exploration with preoperative CTA was observed. The flap survival and occurrence of complications were observed after surgery. The color, appearance, texture, and occurrence of complications were followed up. At the last follow-up, the blood supply of flaps was evaluated using the blood circulation evaluation indicators of Chinese Medical Association Hand Surgery Branch's trial criteria for digital replantation function evaluation, and the function of knee joint was evaluated using knee joint scoring system of hospital for special surgery. Results: The flap condition of the intraoperative exploration was completely consistent with that of preoperative CTA. The flaps survived completely after surgery in 6 patients, while necrosis at the edge of the flap occurred in 1 patient, which healed after dressing change. All the flaps were hyperperfused after surgery, and the color of the flaps gradually became normal after 1 week. Follow-up of 7 to 44 months showed that the color, appearance, and texture were well in all the patients, while local osteomyelitis at the proximal tibia occurred in 1 patient. At the last follow-up, all the 7 patients had excellent blood circulation; the function score of knee joint was 69 to 91, which was evaluated as excellent in 3 cases, good in 3 cases, and fair in 1 case. Conclusions: The retrograde anterolateral thigh flap has large variations, and the application of precise flap surgery method can accurately understand the variations before surgery, guide the design and cutting of the flaps, thus achieving precise repair of anterior knee joint wounds, with good repair outcome.

Clinicopathological characteristics and outcomes of male lupus nephritis in China.

OBJECTIVE: The objective of this article is to assess clinicopathological characteristics and outcomes of patients with male lupus nephritis in a cohort of Chinese patients. METHODS: Clinical, pathological and outcome data of lupus nephritis patients with different gender were retrospectively analyzed and compared. RESULTS: Among 315 patients with renal biopsy-proven lupus nephritis, 45 were male and 270 were female. The average ages of disease onset of the male and female patients were comparable. The interval between presentation of lupus nephritis and diagnosis was significantly longer in the male group than in female group (p = 0.003). Clinical presentation was similar except that males had a significantly lower proportion of alopecia (p = 0.005). In laboratory data, male lupus nephritis patients had higher hemoglobin (p = 0.023) and higher serum creatinine (p < 0.001) than female patients. As for pathological classification and index, no significant difference was found between the two groups. The male patients presented with significantly lower ratios of complete remission and partial remission, and higher ratios of treatment failure and relapse than the female group. Regarding long-term survival and renal outcome, male patients had significantly worse prognosis than females (p = 0.005). CONCLUSIONS: The male lupus nephritis presented with later diagnosis, worse renal function, lower remission rate and higher relapse rate compared with female patients. The male patients had significantly higher mortality and poorer renal outcome.

Adsorption of phenanthrene by quaternary ammonium surfactant modified peat and the mechanism involved.

Removal of phenanthrene (PHE) from aqueous solution by adsorption onto quaternary ammonium surfactant modified peat was studied. The results show that surfactant modification enhanced the PHE adsorption capacity of peat. Low temperature and neutral pH favored PHE adsorption. Peat modified with long carbon chain surfactant performed better than peat modified with short carbon chain surfactant. The magnitude of PHE adsorption capacity followed the order of MP-HPB>MP-HTAB>MP-TBAB>RP, ranged from 924 to 1,228 µg g(-1). A negative trend between adsorption capacity (y) and (O+N)/C ratio of biosorbent (x) was observed (y = -1,369.6x + 2,176), which confirmed the negative effect of polarity on polycyclic aromatic hydrocarbon (PAH) removal. The study provides a guide to modify raw materials to enhance adsorption of hydrophobic organics.

Long non-coding RNAs and small nucleolar RNA host gene 6 contribute to the depression-like behavior of hypothyroid mice by promoting methylation of the brain-derived neuropathic factor promoter that is mediated by DNA methyltransferase 1.

Long non-coding RNAs (lncRNAs) and small nucleolar RNA host gene 6 (SNHG6) have attracted extensive attention due to their involvement in various pathological processes. However, the functional role of lncRNA SNHG6 in depression-like behavior induced by hypothyroidism is still largely unknown. Our study was designed to explore the biological role of lncRNA SNHG6 in depression-like behavior induced by hypothyroidism and the underlying mechanisms. First, the depression-like behavior of hypothyroid mice was investigated after lncRNA SNHG6 knockdown. Subsequently, the regulation of the methylation levels of the brain-derived neurotrophic factor (BDNF) promoters by lncRNA SNHG6 was evaluated. To reveal the underlying mechanisms of lncRNA SNHG6 in the methylation of the BDNF promoters, RNA pull-down assays, RNA immunoprecipitation, and co-immunoprecipitation were performed. Further experiments were also conducted to investigate the roles of DNA (cytosine-5)-methyltransferase 1 (DNMT1) in the depression-like behavior induced by hypothyroidism. In this study, elevated levels of lncRNA SNHG6 were noted in the hippocampus in hypothyroid mice. Function assays proved that lncRNA SNHG6 knockdown alleviated the depression-like behavior induced by hypothyroidism. Furthermore, a mechanistic investigation validated that lncRNA SNHG6 stabilized DNMT1 by blocking UHRF1-mediated DNMT1 ubiquitination, which increased the methylation levels of the BDNF promoters. Moreover, DNMT1 was found to be involved in depression-like behavior in hypothyroid mice. Our findings revealed a novel mechanism by which lncRNA SNHG6 promotes the methylation levels of the BDNF promoters by stabilizing DNMT1 and sheds light on potential therapeutic strategies for depression-like behavior induced by hypothyroidism.

[Preparation of CD33 targeted bispecific- and trispecific-T cell engagers and their cytotoxicity on leukemia cells].

Objective: To investigate the effect of CD33-targeted bi-specific and tri-specific T-cell engagers on T-cell proliferation and explore their cytotoxicity on leukemia cells. Methods: The CD33-targeted bi-specific T-cell engager (CD33-BiTE) and tri-specific T-cell engager (CD33-TriTE) expression vectors were successfully constructed and expressed through a eukaryotic cell expression system. CD33-BiTE and CD33-TriTE were purified by affinity chromatography. The effects of CD33-BiTE and CD33-TriTE on T cells were analyzed through in vitro experiments. Results: ① CD33-BiTE and CD33-TriTE were successfully constructed and purified and could compete with flow cytometry antibodies for binding to the target cells. ② After 12 days of co-culture with CD33-BiTE and CD33-TriTE, the number of human T cells were expanded to 33.89±19.46 and 81.56±23.62 folds, respectively. CD33-TriTE induced a stronger proliferation of T cells than CD33-BiTE (P<0.05) . ③ Both CD33-BiTE and CD33-TriTE induced specific dose-dependent cytotoxicity on CD33(+) leukemia cells. ④ Compared to CD33-TriTE, leukemia cells were prone to express PD-L1 when co-cultured with T cells and CD33-BiTE. CD33-TriTE induced powerful cytotoxicity on leukemia cells with high PD-L1 expression. Conclusion: CD33-BiTE and CD33-TriTE expression vectors were constructed, and fusion proteins were expressed in eukaryotic cells. Our results support the proliferative and activating effects of BiTE and TriTE on T cells. Compared to that of CD33-BiTE, CD33-TriTE induced a stronger proliferative effect on T cells and a more powerful cytotoxicity on leukemia cells with high PD-L1 expression.

[Preparation of CD52-targeted chimeric antigen receptor-modified T cells and their anti-leukemia effects].

Objective: To construct chimeric antigen receptor (CAR) T cells targeting CD52 (CD52 CAR-T) and validate the effect of CD52 CAR-T cells on CD52-positive leukemia. Methods: A second-generation CD52-targeting CAR bearing 4-1BB costimulatory domain was ligated into a lentiviral vector through molecular cloning. Lentivirus was prepared and packaged by 293 T cells with a four-plasmid system. Fluorescein was used to label cell surface antigens to evaluate the phenotype of CD52 CAR-T cells after infection. Flow cytometry and ELISA were used to evaluate the specific cytotoxicity of CD52 CAR-T cells to CD52-positive cell lines in vitro. Results: ①A pCDH-CD52scFv-CD8α-4-1BB-CD3ζ-GFP expressing plasmid was successfully constructed and used to transduce T cells expressing a novel CD52-targeting CAR. ②On day 6, CD52-positive T cells were almost killed by CD52-targeted CAR-T post lentivirus transduction [CD52 CAR-T (4.48 ± 4.99) %, vs Vector-T (56.58±19.8) %, P=0.011]. ③T cells transduced with the CAR targeting CD52 showed low levels of apoptosis and could be expanded long-term ex vivo. ④The CD52 CAR could promote T cell differentiation into central and effector memory T cells, whereas the proportion of T cells with a CD45RA(+) effector memory phenotype were reduced. ⑤CD52 CAR-T cells could specifically kill CD52-positive HuT78-19t cells but had no killing effect on CD52-negative MOLT4-19t cells. For CD52 CAR-T cells, the percentage of residual of HuT78-19t cells was (2.66±1.60) % at an the E:T ratio of 1∶1 for 24 h, while (56.66±5.74) % of MOLT4-19t cells survived (P<0.001) . ⑥The results of a degranulation experiment confirmed that HuT78-19t cells significantly activated CD52 CAR-T cells but not MOLT4-19t cells[ (57.34±11.25) % vs (13.06± 4.23) %, P<0.001]. ⑦CD52 CAR-T cells released more cytokines when co-cultured with HuT78-19t cells than that of vector-T cells [IFN-γ: (3706±226) pg/ml, P<0.001; TNF-α: (1732±560) pg/ml, P<0.01]. Conclusions: We successfully prepared CD52 CAR-T cells with anti-leukemia effects, which might provide the foundation for further immunotherapy.

[Functional investigation of chimeric antigen receptor T cells targeting LMP1 antigen].

Objective: This study aimed to create a type of CAR-T cells that targets LMP1 antigen and study its immunotherapeutic effect on LMP1-positive hematological malignancies. Methods: To generate LMP1 CAR-T cells, a plasmid expressing LMP1 CAR was created using molecular cloning technology, and T cells were infected with LMP1 CAR lentivirus. The effects of LMP1 CAR-T cells on specific cytotoxicity against LMP1-positive tumor cell lines infected with the EB virus had been confirmed. Results: ① LMP1 protein expressing on EB virus-positive lymphoma cells surface was verified. ② The LMP1 CAR-expressing plasmid was created, and LMP1 CAR-T cells were obtained by infecting T cells with a lentivirus packaging system, with an infection efficiency of more than 80% . ③LMP1 CAR-T cells have a 4∶1 effect-to-target ratio in killing LMP1-positive lymphoma cells. The killing effect of LMP1 CAR-T cells on Raji cells was enhanced after 48 h of coculture, but there was no significant killing effect on Ramos, which are LMP1-negative lymphoma cells. ④After coculture with LMP1-positive lymphoma cells at a ratio of 1∶1 for 5 h, the degranulation effect was enhanced. The proportion of CD107a(+) T cells in the LMP1 CAR-T cell treatment group was significantly higher than that in the vector-T cell group [ (13.25±2.94) % vs (1.55±0.05) % , t=3.972, P=0.017]. ⑤After coculture with LMP1-positive lymphoma cells, the proportion of CD69(+) and CD25(+) T cells in the LMP1 CAR-T cell group was significantly higher than that in vector-T cell group [ (7.40±0.41) % vs (3.48±0.47) % , t=6.268, P=0.003; (73.00±4.73) % vs (57.67±2.60) % , t=2.842, P=0.047]. ⑥After coculture with LMP1-positive lymphoma cells, cytokine secretion in the LMP1 CAR-T cell group was higher than that in the vector-T cell group [interferon-gamma: (703±73) ng/L vs (422±87) ng/L, t=2.478, P=0.068; tumor necrosis factor-alpha: (215±35) ng/L vs (125±2) ng/L, t=2.536, P=0.064]. Conclusion: In this study, we found that the LMP1 protein is only found on the surface of the EBV-positive tumor cell. Simultaneously, we created an LMP1 CAR-expressing plasmid and obtained LMP1 CAR-T cells by infecting T cells with a lentivirus packaging system. Furthermore, we demonstrated that LMP1 CAR-T cells could specifically kill LMP1-positive tumor cells in vitro. The degranulation and activation effects of LMP1 CAR-T cells were enhanced after coculture with LMP1-positive tumor cells, indicating a potential clinical application.