High MCM6 expression promotes proliferation and correlates with poor prognosis in triple-negative breast cancer.

OBJECTIVE: Triple-negative breast cancer (TNBC) is an aggressive subtype with a poor prognosis. Minichromosome maintenance genes (MCM2-7) crucial for DNA replication are significant biomarkers for various tumor types; however, their roles in TNBC remain underexplored. MATERIALS AND METHODS: We utilized four TNBC-related GEO databases to examine MCM2-7 gene expression and predict its prognosis in TNBC, performing single-cell analysis and GSEA to discover MCM6's potential function. The Cancer Dependency Map gene effect scores and CCK8 assay were used to assess MCM6's impact on TNBC cell proliferation. The correlations between MCM6 expression, immune infiltrates, and immune cells were also analyzed. WGCNA and LASSO Cox regression built a risk score model predicting TNBC patient survival based on MCM6-related gene expression. RESULTS: MCM2-7 gene expression was higher in TNBC tissues compared to adjacent normal tissues. High MCM6 expression correlated with shorter TNBC patient survival time. GSEA and single-cell analysis revealed a relationship between elevated MCM6 expression and the cell cycle pathway. MCM6 knockdown inhibited TNBC cell proliferation. A risk model featuring MCM6, CDC23, and CCNB1 effectively predicts TNBC patient survival. CONCLUSIONS: MCM6 overexpression in TNBC links to a worse prognosis and reduced cell proliferation upon MCM6 knockdown. We developed a risk score model based on MCM6-related genes predicting TNBC patient prognosis, potentially assisting future treatment strategies.

Laser vibrational excitation of radicals to prevent crystallinity degradation caused by boron doping in diamond.

Pursuing high-level doping without deteriorating crystallinity is prohibitively difficult but scientifically crucial to unleashing the hidden power of materials. This study demonstrates an effective route for maintaining lattice integrity during the combustion chemical vapor deposition of highly conductive boron-doped diamonds (BDDs) through laser vibrational excitation of a growth-critical radical, boron dihydride (BH2). The improved diamond crystallinity is attributed to a laser-enabled, thermal nonequilibrium suppression of the relative abundance of boron hydrides (BH), whose excessive presence induces boron segregation and disturbs the crystallization. The BDDs show a boron concentration of 4.3 × 1021 cm-3, a film resistivity of 28.1 milliohm·cm, and hole mobility of 55.6 cm2 V-1 s-1, outperforming a commercial BDD. The highly conductive and crystalline BDDs exhibit enhanced efficiency in sensing glucose, confirming the advantages of laser excitation in producing high-performance BDD sensors. Regaining crystallinity with laser excitation in doping process could remove the long-standing bottlenecks in semiconductor industry.

Urinary microRNAs miR-15b and miR-30a as novel noninvasive biomarkers for gentamicin-induced acute kidney injury.

MicroRNAs serve as potential biomarkers in various pathological models, and are stable and detectable in biofluids. We investigated the urinary microRNA expression profile in a gentamicin-induced acute kidney injury canine model using RNA sequencing. A total of 234 differentially expressed microRNAs were screened after 12 consecutive days of gentamicin administration (P < 0.05). Six candidate microRNAs (miR-15b, -15b-3p, -16, -30a, -30a-3p, and -30c-2-3p) were selected according to a set criterion, and validated by real-time quantitative PCR. The diagnostic values of these six candidate microRNAs were better than the traditional serum biomarkers (all P < 0.05). Further, using receiver operating characteristic curve analysis, we found that miR-15b and -15b-3p were superior to urinary kidney injury molecule-1 (both P < 0.05). Moreover, miR-15b and -30a levels in the urine samples significantly correlated with their respective levels in the kidney tissue samples (r=0.512 and 0.505, respectively, both P < 0.05). Our data concluded that miR-15b and -30a may be promising biomarkers for renal toxicity.

[Analysis on the cluster epidemic of coronavirus disease 2019 in Guangdong Province].

Objective: Analysis of clustering characteristics of coronavirus disease 2019 (COVID-19) in Guangdong Province. Methods: The COVID-19 cases in Guangdong Province onset from January 1 to February 29, 2020 were collected from Chinese information system for disease control and prevention and Emergency Public Reporting System. Obtain the epidemiological survey data of the cluster epidemic situation, and clarify the scale of cluster epidemic situation, the characteristics of the index cases, family and non-family subsequent cases. Calculate serial interval according to the onset time of the index cases and subsequent cases, secondary attack rate based on the close contacts tracking results, the characteristics of different cases in the clustered epidemic were compared. Results: A total of 283 cluster were collected, including 633 index cases, 239 subsequent cases. Families are mainly clustered, the total number involved in each cluster is in the range of 2-27, M (P25, P75) are 2.0 (2.0, 4.0). During January 15 to February 29, the secondary attack rate is 2.86% (239/8 363) in Guangdong Province, the family secondary attack rate was 4.84% (276/3 697), and the non-family secondary attack rate was 1.32% (61/4 632). According to the reporting trend of the number of cases in Guangdong Province, it can be divided into four stages, the rising stage, the high platform stage, the descending stage and the low level fluctuation period. The secondary attack rate of the four stages were 3.5% (140/3 987), 2.3% (55/2 399), 2.6% (37/1 435), 1.3% (7/542), respectively. The difference was statistically significant (P=0.003). Conclusion: COVID-19 cluster mainly occurs in families in Guangdong Province. The scale of the clustered epidemic was small; the serial interval was short; and the overall secondary attack rate was low.

[Clinical outcomes of COVID-19 cases and influencing factors in Guangdong province].

Objective: To analyze the clinical courses and outcomes of COVID-19 cases and the influencing factors in Guangdong province and provide basis for the formulation or adjustment of medical care and epidemic control strategy for COVID-19. Methods: We collected demographic data, medical histories, clinical courses and outcomes of 1 350 COVID-19 patients reported in Guangdong as of 4 March 2020 via epidemiological investigation and process tracking. Disease severity and clinical course characteristics of the patients and influencing factors of severe illness were analyzed in our study. Results: Among 1 350 cases of COVID-19 cases in Guangdong, 72 (5.3%) and 1 049 (77.7%) were mild and ordinary cases, 164 (12.1%) were severe cases, 58 (4.3%) were critical cases and 7 (0.5%) were fatal. The median duration of illness were 23 days (P(25), P(75): 18, 31 days) and the median length of hospitalization were 20 days (P(25), P(75): 15,27 days). For severe cases, the median time of showing severe manifestations was on the 12(th) day after onset (P(25), P(75): 9(th) to 15(th) days), and the median time of severe manifestation lasted for 8 days (P(25), P(75): 4, 14 days). Among 1 066 discharged/fetal cases, 36.4% (36/99) and 1.0% (1/99) of the mild cases developed to ordinary cases and severe cases respectively after admission; and 5.2% (50/968) and 0.6% (6/968) of the ordinary cases developed to severe cases, and critical cases respectively after admission. In severe cases, 11.4% developed to critical cases (10/88). The influencing factors for severe illness or worse included male (aHR=1.87, 95%CI: 1.43-2.46), older age (aHR=1.67, 95%CI: 1.51-1.85), seeking medical care on day 2-3 after onset (aHR=1.73, 95%CI: 1.20-2.50) pre-existing diabetes (aHR=1.75, 95%CI: 1.12-2.73) and hypertension (aHR=1.49, 95%CI: 1.06-2.09). Conclusions: The course of illness and length of hospitalization of COVID-19 cases were generally long and associated with severity of disease clinical outcomes. The severe cases were mainly occurred in populations at high risk. In the epidemic period, classified management of COVID-19 cases should be promoted according to needs for control and prevention of isolation and treatment for the purpose of rational allocation of medical resources.

[Correlation between CTL and Th1 cell proportion in peripheral blood of liver transplantation recipients and the success of hepatitis B vaccination].

Objective: To investigate the correlation between the proportion of CTL and Th1 cells in peripheral blood of liver transplant recipients and the success of hepatitis B vaccination. Methods: The subjects of this study were liver transplantation recipients with chronic HBV-related liver diseases in Organ transplantation institute of the third medical center of PLA general hospital. Subjects were randomly divided into two groups for prospective study. In the rapid group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2and 3 months, and one dose of 20 µg hepatitis B vaccine was administered at 4, 5 and 6 months. In the rapid-enhanced group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2 and 3 months, and one dose of 60 µg hepatitis B vaccine was administered at 4, 5 and 6months. Compare and analyze the success rate of inoculation, the titer of hepatitis B surface antibody (anti-HBs), the proportion of CTL cells in CD8(+)T cells and Th1 cells in CD4(+)T cells. Correlation analysis was performed for CTL and Th1 cells and anti-HBs, Observe the safety of vaccination. Results: The inoculation success rate, anti-HBs growth rate, CTL cell percentage increase and Th1 cell percentage increase in the rapid enhancement group were all higher than those in the rapid enhancement group, and the differences were statistically significant, they were 38.3% (23/60) vs 21.7% (13/60) (P=0.046), 91.3(72.5,124.2) vs 22.1(12.4, 31.6) (P=0.001), 1.4(0.8,1.9) vs 0.4(0.2,1.4) (P=0.001) and 7.4±2.6 vs 5.6±3.7 (P=0.001) respectively. The percentage increase of CTL cells and Th1 cells in the successful group was greater than that in the non-successful group, and the difference was statistically significant. They were 1.9(1.4,2.5) vs 0.1(0.0,1.1) (P=0.024) and 9.6±3.1 vs 2.4±2.0 (P<0.001). There was no significant correlation between anti-HBs increase (105.5±37.1) and CTL increase 1(0,3) (P=0.099), while there was significant positive correlation with Th1 increase 7(2,11) (P<0.001). No rejection reaction occurred during the study period, and there was no special abnormal change in the safety index. Conclusion: Reasonable increase of vaccine dose can up-regulate Th1 cell expression and promote the generation of anti-HBs.

[Overexpression of protein phosphatase 2 regulatory subunit B''α gene effect on proliferation and invasion of hepatoma cells].

Objective: To study the overexpression of protein phosphatase 2 regulatory subunit B''α gene effects on the proliferation and invasion of hepatoma cells. Methods: Immunohistochemistry method was used to analyze the expression of PPP2R3A in cancerous and paracancerous tissues. Hepatocellular carcinoma cell lines (Huh-7 and HepG2) with stably overexpressing PPP2R3A were constructed by lentiviral vector. Biological behavioral transition in hepatocellular carcinoma cell proliferation, cell cycle, apoptosis, invasion and metastasis were detected by cell counting kit-8 assay (CCK-8), flow cytometry, and transwell assay. A subcutaneous nude tumor mice model was constructed to validate the growth of hepatoma cells. Two independent sample t-tests were used to compare the groups. Results: The expression of PPP2R3A gene in human hepatocarcinoma tissues was higher than paracancerous tissues. The absorbance (A value) of hepatoma cells was increased (P < 0.05) after overexpression of PPP2R3A gene. The transition from G1-to-S phase was significantly increased i.e., the G1 phase of the cell cycle was reduced (Huh-7: t = 3.04, P = 0.0384; HepG2: t = 4.06, P = 0.0153), while the S phase was increased (Huh-7: t = 3.47, P = 0.0255; HepG2: t = 4.46, P = 0.0112). Early apoptotic rate was decreased (Huh-7: t = 7.34, P = 0.0018; HepG2: t = 4.06, P = 0.0153). The number of Huh-7 cells migrating to the lower chamber was increased (t = 3.18, P = 0.0334), and after the use of matrigel the number of cells reaching to the lower chamber was also increased (t = 2.84, P = 0.0464). The results of animal experiments showed that the subcutaneous tumor growth (t = 4.31, P = 0.0035) was significantly overexpressed in nude mice group. The results of Western blot showed that the expression of PARP and P53 protein in the spliced ​​forms decreased, while the accumulation of ß-catenin protein in the liver cancer cells was increased. Conclusion: Overexpressed PPP2R3A gene may promote proliferation, migration and invasion ability, inhibit apoptosis, induce G1/S phase transition, and participate in the biological behavior of hepatoma cells.

[Overview of traditional Chinese medicine of "Xiang Qi" theory of origins].

The theory of "Xiang Qi" (inspecting Qi or inspecting pneumata) is an important method of TCM inspection.It has important value in clinical diagnosis.The theoretical framework of "Xiang Qi" has been formed from the Huangdi Neijing (The Yellow Emperor's Classic of Medicine)《》. The many ideas were improved and developed in many dynasties such as Sui, Tang, Song, Jin and Yuan. Especially the theory of "Xiang Qi" has been inherited and applied in Ming and Qing dynasties, the theory of "Xiang Qi" became an important diagnostic method for the clinical practice.Especially Wang Hong()'s Wang Zhen Zun Jing(Inspection following the classics)《》 in Qing Dynasty was published.It perfected and developed the ten kinds of methods of "Xiang Qi" from three aspects. And it contained four diagnostic methods by examining Qi to determine the disease.It has promoted the value and the clinical application of "Xiang Qi" theory, and has contributed to the development of "Xiang Qi" theory.

[Biomechanical study of different kinds of internal fixation for the typeâ Hangman fracture, type â ¡ odontoid fracture and the C(2/3) disc injury: a finite-element analysis].

Objectives: To analyze the biomechanical stability of four kinds of internal fixation for the type Ⅰ Hangman fracture, type Ⅱ odontoid fracture and the C(2/3) disc injury by finite element (FE) analysis. Methods: Thin-section spiral computed tomography (0.5 mm) was performed on C(1) to C(3) region of cervical vertebra in healthy male volunteers.A three-dimensional hexahedral FE model of upper cervical spine was established by software (Mimics, GEOMAGICS, Pro/E and Ansys). Then the weakening of the strength of grid was performed to simulate the FE model of the type Ⅰ Hangman fracture, type Ⅱ odontoid fracture and the C(2/3) disc injury (FE/Fracture), the four internal fixation models: anterior cervical plate+ odontoid screw+ cage (FE/ACP+ OS+ cage), affixing rods from pedicle screws in C(2) to lateral mass screws in C(3)+ odontoid screw + cage (FE/C(2)PS+ C(3)LMS+ OS+ cage), affixing rods from pedicle screws in C(1) to pedicle screws in C(2) and lateral mass screws in C(3) (FE/C(1)PS+ C(2)PS+ C(3)LMS), anterior odontoid screw plate fixation system (FE/AOSP) were simulated on the FE/Fracture model.Flexion, extension, lateral bending and axial rotation were imposed on the FE/Intact, FE/Fracture and the four fixation models respectively. Results: The intact model of upper cervical spine (C(1)-C(3)) was established successfully, consisting of 259 641 nodes and 403 674 units.There was no significant difference among the FE/ACP+ OS+ cage, the FE/ C(2)PS+ C(3)LMS+ OS+ cage and the FE/AOSP of ROMC(1/2).During flexion, extension, left axial rotation and right axial rotation of ROMC(2)-C(3), the FE/AOSP decreased 70.7%, 74.4%, 38.9%, 41.1% respectively compared with the FE/C(1)PS+ C(2)PS+ C(3)LMS.The ROMC(2)-C(3) during flexion, extension, left lateral bending, right lateral bending, left axial rotation and right axial rotation in the FE/AOSP decreased for 82.2%, 82.8%, 73.2%, 64.8%, 72.2%, 81.5% respectively when compared with those in FE/ACP+ OS+ cage.The ROMC(2)-C(3) during flexion, extension, left lateral bending, right lateral bending, left axial rotation and right axial rotation in the FE/AOSP decreased 88.2%, 81.2%, 47.6%, 41.2%, 38.9%, 39.0% respectively when compared with those in FE/C(2)PS+ C(3)LMS+ OS+ cage.The stress concentrated on the connection between plate and screw in the FE/ACP+ OS+ cage, the FE/C(2)PS+ C(3)LMS+ OS+ cage and the FE/C(1)PS+ C(2)PS+ C(3)LMS, while it distributed evenly in the FE/AOSP. Conclusion: Anterior odontoid screw plate fixation system can be used to treat the type Ⅰ Hangman fracture, type Ⅱ odontoid fracture, and the C(2/3) disc injury and can reserve the function of atlanto-axial joint.

[Clinical analysis of de novo HBV infection after liver transplantation in non-HBV- related liver disease].

Objective: The aim of this study was to summarize and analyze the clinical features and characteristics of de novo HBV infection after liver transplantation in non-HBV-related liver disease. Methods: We retrospectively analyzed the clinical data of 13 patients with new HBV infection in 376 cases of liver transplantation patients with non-HBV related liver diseases from April 2002 to December 2013 in our hospital. Results: Among 376 patients with non-HBV-related liver disease after liver transplantation, 13 patients developed new HBV infection, and the rate of new HBV infection was 3.46%. Of the 13 cases, 5 were males and 8 were females. The follow-up time was 14.7 -128.7 months, and the average time from surgery to new HBV infection was 19.06 months. The primary diseases were as follows: 5 cases of primary biliary cholangitis, 3 cases of alcoholic liver disease, 2 cases of drug-induced liver damage, 1 cases of post-hepatitis C cirrhosis, congenital biliary atresia and congenital liver fibrosis. All patients were positive for HBsAg, HBeAg, anti-HBc, 11 were positive for HBV DNA, and 2 were negative for HBV DNA. 6 cases had abnormal liver function and 7 cases had normal liver function. All patients were treated with antiviral therapy with nucleoside (acid) analogues. HBsAg was negative in 6 patients; HBsAg remained positive in 7 cases, including HBsAg, HBeAg, anti-HBc positive in 6 cases, HBsAg, anti-HBe, anti- HBc was positive in 1 case, HBV DNA was still positive in 1 patient, and HBV DNA was negative in 6 patients; liver function was normal in all patients. Conclusion: Non-HBV- related liver transplantation are high-risk group of new HBV infection, with the highest incidence of autoimmune liver disease. It is speculated that it may be related to the long-term use of hormones after the transplantation. The prognosis of newly diagnosed HBV infection after liver transplantation is fine as long as it can be found and treated early.

Enhancement or inhibition of PLCγ2 expression in rat hepatocytes by recombinant adenoviral vectors that contain full-length gene or siRNA.

We investigated the effects of recombinant adenovirus vectors that overexpress or silence PLCγ2 on the expression of this gene during hepatocyte proliferation. Hepatocytes were isolated, identified by immunofluorescent cytochemical staining and infected by previously constructed Ad-PLCγ2 and Ad-PLCγ2 siRNA1, siRNA2 and siRNA3. Green fluorescent protein (GFP) expression was observed by fluorescence microscopy. Infection percentage was calculated by flow cytometry. mRNA and protein levels of PLCγ2 were detected by quantitative reverse transcription-PCR (qRT-PCR) and western blotting, respectively. The viability of the infected hepatocytes was measured by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. We found that nearly 97% of cells were positive for the hepatocyte marker, CK18. After infection of Ad-PLCγ2 and Ad-PLCγ2 siRNA, more than 99% of hepatocytes expressed GFP significantly, and mRNA and protein expression of PLCγ2 was up-regulated significantly in Ad-PLCγ2 infected hepatocytes, but down-regulated in Ad-PLCγ2 siRNA2 infected cells. The cell proliferation rate decreased in PLCγ2-overexpressing cells, while the rate increased in PLCγ2-silencing cells. We verified that recombinant Ad-PLCγ2 and Ad-PLCγ2 siRNA2 were constructed successfully. These two recombinant vectors promoted or decreased the expression of PLCγ2 in rat hepatocytes and affected the cell proliferation rate, which provides a useful tool for further investigation of the role of PLCγ2 in hepatocyte apoptosis.

Genome-wide analysis of salinity-stress induced DNA methylation alterations in cotton (Gossypium hirsutum L.) using the Me-DIP sequencing technology.

Cytosine DNA methylation is a significant form of DNA modification closely associated with gene expression in eukaryotes, fungi, animals, and plants. Although the reference genomes of cotton (Gossypium hirsutum L.) have been publically available, the salinity-stress-induced DNA methylome alterations in cotton are not well understood. Here, we constructed a map of genome-wide DNA methylation characteristics of cotton leaves under salt stress using the methylated DNA immunoprecipitation sequencing method. The results showed that the methylation reads on chromosome 9 were most comparable with those on the other chromosomes, but the greatest changes occurred on chromosome 8 under salt stress. The DNA methylation pattern analysis indicated that a relatively higher methylation density was found in the upstream2k and downstream2k elements of the CDS region and CG-islands. Almost 94% of the reads belonged to LTR-gspsy and LTR-copia, and the number of methylation reads in LTR-gypsy was four times greater than that in LTR-copia in both control and stressed samples. The analysis of differentially methylated regions (DMRs) showed that the gene elements upstream2k, intron, and downstream2k were hypomethylated, but the CDS regions were hypermethylated. The GO (Gene Ontology) analysis suggested that the methylated genes were most enriched in cellular processes, metabolic processes, cell parts and catalytic activities, which might be closely correlated with response to NaCl stress. In this study, we completed a genomic DNA methylation profile and conducted a DMR analysis under salt stress, which provided valuable information for the better understanding of epigenetics in response to salt stress in cotton.

Quality assessment of groundwater from the south-eastern Arabian Peninsula.

Assessment of groundwater quality plays a significant role in the utilization of the scarce water resources globally and especially in arid regions. The increasing abstraction together with man-made contamination and seawater intrusion have strongly affected groundwater quality in the Arabia Peninsula, exemplified by the investigation given here from the United Arab Emirates, where the groundwater is seldom reviewed and assessed. In the aim of assessing current groundwater quality, we here present a comparison of chemical data linked to aquifers types. The results reveal that most of the investigated groundwater is not suitable for drinking, household, and agricultural purposes following the WHO permissible limits. Aquifer composition and climate have vital control on the water quality, with the carbonate aquifers contain the least potable water compared to the ophiolites and Quaternary clastics. Seawater intrusion along coastal regions has deteriorated the water quality and the phenomenon may become more intensive with future warming climate and rising sea level.

Comparison of Mizoribine and Mycophenolate Mofetil With a Tacrolimus-Based Immunosuppressive Regimen in Living-Donor Kidney Transplantation Recipients: A Retrospective Study in China.

BACKGROUND: A retrospective study was conducted to investigate the prevalence of gastrointestinal (GI) symptoms as well as the efficacy and safety of mizoribine (MZR) and mycophenolate mofetil (MMF) in Chinese living-donor kidney transplantation (LDKT). METHODS: Forty-two recipients enrolled between January 2012 and March 2014 were treated with either MZR (n = 22) or MMF (n = 20). All patients were treated in combination with a tacrolimus-based immunosuppressive regimen, besides the study drugs. RESULTS: GI symptoms were observed in 1 of 22 patients (4.5%) and 10 of 20 patients (50%) in MZR treatment group and MMF treatment group, respectively (P = .001), during the post-transplantation 1 year. No significant differences in the incidence of acid reflux, bloated stomach feeling, and constipation were observed between the two groups. No recipient developed diarrhea in the MZR treatment group, whereas 30% of the MMF treatment group developed diarrhea (P = .007). The averages of GI symptom severity total score and diarrhea score were significantly lower in the MZR treatment group compare with MMF treatment group. There were no inter-group differences in background characteristics. There were no significant differences in acute rejection rate and clinical findings between these two groups, whereas the prevalence of cytomegalovirus infection and leukopenia were significantly lower in the MZR treatment group. There was no significant difference on adverse events such as hyperuricemia or other adverse events. CONCLUSIONS: This study demonstrated a significantly lower incidence of GI symptoms for treatment with MZR compared with MMF and good efficacy and safety in Chinese LDKT with MZR.

[Risk factors for hepatitis B recurrence after liver transplantation for HBV-related liver disease].

Objective: To investigate the risk factors for hepatitis B recurrence after liver transplantation for HBV-related liver disease, and to provide a basis for effective preventive measures. Methods: A retrospective analysis was performed for the clinical data of 907 patients with HBV-related liver disease who underwent liver transplantation from April 2002 to December 2013. The chi-square test was used to determine the risk factors for hepatitis B recurrence, and multivariate Cox regression analysis was used for identifying the independent risk factors. Results: There were 907 patients with HBV-related liver disease who underwent liver transplantation. The patients were followed up for 1-144 months, with a median follow-up time of 50.9 months. Among them, 56 experienced hepatitis B recurrence, yielding a recurrence rate of 6.17%. The univariate analysis showed that positive HBeAg before surgery, positive HBV DNA before surgery, positive anti-HBc in liver donor, postoperative tumor recurrence, and postoperative regimen for the prevention of hepatitis B recurrence were significantly correlated with hepatitis B recurrence after liver transplantation (P < 0.05). The multivariate Cox regression analysis showed that positive HBeAg before surgery (OR = 1.891, 95% CI 1.064-3.360, P < 0.05), positive anti-HBc in liver donor (OR = 3.128, 95% CI 1.591-6.151, P < 0.01), and postoperative tumor recurrence (OR = 4.365, 95% CI 2.152-8.857, P < 0.01) were independent risk factors for hepatitis B recurrence. Conclusion: Positive HBeAg before surgery, postoperative tumor recurrence, and positive anti-HBc in liver donor are independent risk factors for hepatitis B recurrence after liver transplantation. For the patients who plan to undergo liver transplantation, antiviral therapy should be given before surgery to reduce HBV viral load, and effective preventive measures after surgery are the key to the prevention and reduction of postoperative hepatitis B recurrence.

[Zheng Meijian's Sha yu yi yu (Random Medical Talks in Leisure Time) and his new ideas on pulse-taking].

Zheng Meijian, a doctor of Xin'an district in the reign of Qianlong Emperor of the Qing Dynasty, left his manuscript Sha yu yi yu (Random Medical Talks in Leisure Time), and was handed down in his family for generations. Sha yu yi yu was published in the Anhui Yixue of Issue 20, 1959, with horizontal layout and punctuation. The book records Zheng Meijian's understanding for the medicine, especially his new ideas on the cunkou pulse, with its underlying corresponding relationship among cun-guan-chi and zang-fu which should be combined with meridian and collateral system, claiming that the pulse taken superficially was corresponding to fu, while that taken deeply, to zang. He classified the strength of the fingers in pulse-taking into 8 levels, and advocated flexible application of pulse-taking. All his opinions had an important reference value for theoretical research and clinical application of pulse-taking.

Construction of recombinant adenovirus Ad-rat PLCg2-shRNA and successful suppression of PLCg2 expression in BRL-3A cells.

Phospholipase Cg2 (PLCg2) induces apoptosis of immune and tumor cells; however, it remains unclear whether PLCg2 promotes hepatocyte apoptosis during liver regeneration (LR). Therefore, to establish a framework for further exploring the function of PLCg2, we generated recombinant adenoviruses carrying a template encoding short hairpin (sh)-RNA targeting PLCg2 (Ad-PLCg2-shRNA), which were used to silence the expression of PLCg2 in BRL-3A cells. First, three pairs of PLCg2-shRNAs were designed, synthesized, and cloned into a shuttle vector, pHBAd-U6-GFP, after annealing. The recombinant shuttle plasmids were co-transfected with the backbone vector pHBAd-BHG into HK293 cells to package the recombinant Ad-PLCg2-shRNAs used to infect BRL-3A cells. Infection efficiency was monitored by observing the number of GFP-positive cells under a fluorescent microscope. To determine the recombinant adenoviruses with the highest silencing efficiency, levels of PLCg2 mRNA were evaluated by qRT-PCR. DNA sequencing confirmed that the correct shRNA coding sequences were inserted into the shuttle vectors and adenoviral plasmids. The titers of three recombinant adenoviruses were at least 1 x 10(10) PFU/mL. The most effective adenoviral construct, with interference efficiency of 77%, was determined by qRT-PCR. These results show that a recombinant adenovirus, Ad-PLCg2-shRNA, was developed and was effective at silencing the rat PLCg2 gene. This construct may contribute to the study of PLCg2 in hepatocyte apoptosis during LR.