[Comparison of clinical efficacy of robotic, laparoscopic and open surgery in the treatment of severe rectal prolapse].

Objective: To analyze and compare the efficacy of robotic, laparoscopic and open dorsal mesh rectopexy in the treatment of severe rectal prolapse. Methods: A retrospective cohort study was performed. Patients who had a full-thickness rectum pulled out of the anus before surgery and the length was greater than 8 cm, and underwent transabdominal dorsal mesh rectopexy were enrolled in the study. Those who had urinary or sexual dysfunction before surgery, could not perform sexual function scores due to lack of a fixed sexual partner or sexual activity after surgery, underwent laparotomy again during the perioperative period, were transferred to laparotomy during robotic or laparoscopic surgery, or had no complete information, were excluded. A total of 61 patients with severe rectal prolapse in the First Affiliated Hospital of Zhengzhou University from 2014 to 2018 were enrolled and divided into robotic group (20 cases), laparoscopic group (20 cases) and open group (21 cases) according to the operative procedure based on patients' will. Perioperative parameters were compared among the 3 groups. The International Prostatic Symptoms Score Scale (IPSS, higher score indicates more severe urinary dysfunction), the International Index of Erectile Function questionnaire (IIEF-15, lower score indicates more severe male sexual dysfunction) and the Female Sexual Function Index (FSFI-19, lower score indicates more severe female sexual dysfunction) were used to evaluate and compare the urinary and sexual function before and after operation. Results: There were no significant differences in baseline data among the 3 groups (all P>0.05). In the robotic, laparoscopic and open groups respectively, the operative time was (176.3±13.8) minutes, (160.2±12.1) minutes and (134.2±12.1) minutes; intraoperative blood loss was (58.5±18.9) ml, (67.9±15.7) ml and (114.2±8.4) ml; the first time to ambulation was (19.9±6.8) hours, (24.0±8.9) hours and (37.7±11.4) hours; the first time to gas passage was (31.8±6.8) hours, (35.7±8.9) hours and (49.2±11.2) hours; the hospitalization time was (11.0±1.4) days, (11.4±1.4) days and (13.3±2.1) days; whose differences among 3 groups were all significant (all P<0.001). While no significant differences in morbidity of complication and recurrence among 3 groups were observed (all P>0.05). In the robotic, laparoscopic and open groups respectively, the preoperative IPSS score was (4.2±1.7), (4.4±1.3), and (4.7±1.8); the IPSS score at postoperative 3-month was (8.5±2.5), (9.9±1.7), and (12.2±3.1); IPSS score at postoperative 12-month was (4.3±1.6), (5.8±1.3), and (6.3±1.5), respectively. Compared to preoperative score, postoperative IPSS score increased obviously, then decreased gradually (P<0.001). Preoperative male IIEE score was (22.8±1.8), (22.1±2.1), and (22.6±1.5). In the robotic, laparoscopic and open groups respectively, male IIEE score at postoperative 6-month was (19.6±2.1), (17.1±2.1), and (15.0±2.1); male IIEE score at postoperative 12-month was (22.4±1.6), (19.9±1.5), (17.9±1.8), respectively. Preoperative female FSFI score was (26.4±3.4), (26.6±3.2), and (26.6±3.0); female FSFI score at postoperative 6-month was (21.5±3.3), (18.9±2.9), (17.0±2.6); female FSFI score at postoperative 12-month was (26.1±2.7), (22.7±3.2), and (21.2±2.3), respectively. Postoperative male IIEE score and female FSFI score decreased significantly and then increased gradually with time, whose differences were all significant (all P<0.05). Postoperative IPSS, IIEE, and FSFI scores in the robotic group were superior to those in the laparoscopic and open groups (all P<0.05). Conclusion: Robotic surgery is safe and effective in the treatment of severe rectal prolapse, and is more advantageous in preserving urinary function and sexual function.

Effect of miR-26a on diabetic rats with myocardial injury by targeting PTEN.

OBJECTIVE: To investigate the effect of micro ribonucleic acid (miR)-26a on diabetes-induced myocardial injury in rats by targeting the gene of phosphate and tension homology detected on chromosome ten (PTEN). MATERIALS AND METHODS: Male Wistar rats aged 8-9 weeks old were divided into the control group (n=10), GK group (n=10), and miR-26a agomir group (n=10) according to the body weight. MiRanda and TargetScan target gene prediction software were used to predict and analyze the target gene of miR-26a-5p. The expressions of miR-26a and PTEN in the myocardial tissues of the diabetic rats were detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Hematoxylin-eosin (HE) staining was adopted to observe the pathological changes in the myocardial tissues. In addition, the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was conducted to detect myocardial apoptosis, while the expression of PTEN protein was detected via immunohistochemistry, and the protein expressions of PTEN, b-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteinyl aspartate specific proteinase-3 (Caspase-3) were tested by Western blotting. RESULTS: TargetScan database analysis results showed that miR-26a-5p and PTEN 3'UTR had 6 pairs of complementary bases with the same sequence. Compared with those in the control group, the messenger RNA (mRNA) expression of PTEN in the GK group was notably increased (p<0.05), while that of miR-26a was substantially reduced (p<0.05). In comparison with those in the GK group, the mRNA expression of PTEN was significantly decreased, but that of miR-26a was significantly raised in miR-26a agomir group (p<0.05). Through observation under an optical microscope, it was manifested that in the control group, the myocardial fibers were intact with clear texture but no fracture, and the solid necrosis did not appear in myocardial cells. In the GK group, the myocardial fibers were disorderedly arranged and incomplete with an unclear edge and burrs. The myocardial fibers in the miR-26a agomir group were more regular, with less breakage and solid necrosis. According to TUNEL staining results, the TUNEL-stained brown granules in rats in the GK group were remarkably increased, relative to the control group (p<0.05). Compared with the GK group, miR-26a agomir group had markedly decreased the TUNEL-stained brown particles (p<0.05). It was found in immunohistochemical results that PTEN protein was in lighter color after staining in the control group, with a clear myocardial cell stripe structure. Compared with that in control group, PTEN protein in the GK group was in deeper color after staining, and in comparison with that in the GK group, the color of PTEN protein in miR-26a agomir group became significantly lighter. Moreover, the Western blotting results demonstrated that, compared with those in the control group, the Caspase-3 and Bax protein expressions in the GK group were significantly raised, while Bcl-2 protein expression was notably reduced (p<0.05). Besides, in comparison with the GK group, miR-26a agomir group evidently elevated Caspase-3 and Bax protein expressions and a notably increased Bcl-2 protein expression (p<0.05). CONCLUSIONS: We showed that miR-26a can protect against myocardial injury in diabetic rats by regulating PTEN.

Gene expression in major depressive disorder.

The search for genetic variants underlying major depressive disorder (MDD) has not yet provided firm leads to its underlying molecular biology. A complementary approach is to study gene expression in relation to MDD. We measured gene expression in peripheral blood from 1848 subjects from The Netherlands Study of Depression and Anxiety. Subjects were divided into current MDD (N=882), remitted MDD (N=635) and control (N=331) groups. MDD status and gene expression were measured again 2 years later in 414 subjects. The strongest gene expression differences were between the current MDD and control groups (129 genes at false-discovery rate, FDR<0.1). Gene expression differences across MDD status were largely unrelated to antidepressant use, inflammatory status and blood cell counts. Genes associated with MDD were enriched for interleukin-6 (IL-6)-signaling and natural killer (NK) cell pathways. We identified 13 gene expression clusters with specific clusters enriched for genes involved in NK cell activation (downregulated in current MDD, FDR=5.8 × 10(-5)) and IL-6 pathways (upregulated in current MDD, FDR=3.2 × 10(-3)). Longitudinal analyses largely confirmed results observed in the cross-sectional data. Comparisons of gene expression results to the Psychiatric Genomics Consortium (PGC) MDD genome-wide association study results revealed overlap with DVL3. In conclusion, multiple gene expression associations with MDD were identified and suggest a measurable impact of current MDD state on gene expression. Identified genes and gene clusters are enriched with immune pathways previously associated with the etiology of MDD, in line with the immune suppression and immune activation hypothesis of MDD.

Increased RhoA/ROK mRNA expression and function in diabetic vein grafts compared with nondiabetic vein grafts and diabetic arterial grafts.

OBJECTIVE: Aim of the study was to discuss a new mechanism underlying the poor graft patency of GSV from diabetic patients and provide a rationale for selecting suitable grafts in diabetic patients. MATERIALS AND METHODS: The discarded matched RA, IMA, and GSV from 7 diabetics and 7 nondiabetic patients undergoing CABG were collected and tested for their contractile response to phenylephrine (PE) and their relaxation response to fasudil (a inhibitor of Rho-kinase) and used for immunohistochemical and mRNA detection of RhoA/ROK. RESULTS: The relaxation response to fasudil of GSV taken from diabetic patients was markedly increased but the relaxation response to fasudil of IMA and RA from diabetic patients was not. Immunohistochemistry and mRNA expression of RhoA/ROK was significantly increased in GSV from diabetic patients compared to that of IMA and RA from diabetic patients. RhoA/ROK immunohistochemistry and mRNA expression were significantly increased in GSV from diabetic patients compared with GSV from nondiabetic controls. CONCLUSIONS: RhoA/ROK expression and function in GSV from diabetic patients is significantly increased compared with IMA and RA from diabetic patients and GSV from nondiabetic patients. This contributes to a higher incidence of atherosclerosis and a lower long-term patency of GSV from diabetic patients.

Mutation L437P in the 2B domain of keratin 1 causes diffuse palmoplantar keratoderma in a Chinese pedigree.

Diffuse palmoplantar keratoderma (DPPK) is an autosomal dominant genodermatosis characterized by uniform hyperkeratosis of the palm and sole epidermis. This disorder can be caused by mutations in the genes keratin 1, keratin 9, keratin 16, desmoglein 1 and plakoglobin. Here we present a DPPK Chinese pedigree and identify the aetiology as a novel missense mutation, L437P, located in a highly conserved helix motif in domain 2B of KRT1. Functional analysis shows that overexpression of the L437P mutant in cultured cells leads to abnormal intermediate filament networks and filament aggregation. This gain-of-function mutation highlights the role of domain 2B in mediating filament assembly.

ADME evaluation in drug discovery. 5. Correlation of Caco-2 permeation with simple molecular properties.

The correlations between Caco-2 permeability (logPapp) and molecular properties have been investigated. A training set of 77 structurally diverse organic molecules was used to construct significant QSAR models for Caco-2 cell permeation. Cellular permeation was found to depend primarily upon experimental distribution coefficient (logD) at pH = 7.4, high charged polar surface area (HCPSA), and radius of gyration (rgyr). Among these three descriptors, logD may have the largest impact on diffusion through Caco-2 cell because logD shows obvious linear correlation with logPapp (r=0.703) when logD is smaller than 2.0. High polar surface area will be unfavorable to achieve good Caco-2 permeability because higher polar surface area will introduce stronger H-bonding interactions between Caco-2 cells and drugs. The comparison among HCPSA, PSA (polar surface area), and TPSA (topological polar surface area) implies that high-charged atoms may be more important to the interactions between Caco-2 cell and drugs. Besides logD and HCPSA, rgyr is also closely connected with Caco-2 permeabilities. The molecules with larger rgyr are more difficult to cross Caco-2 monolayers than those with smaller rgyr. The descriptors included in the prediction models permit the interpretation in structural terms of the passive permeability process, evidencing the main role of lipholiphicity, H-bonding, and bulk properties. Besides these three molecular descriptors, the influence of other molecular descriptors was also investigated. From the calculated results, it can be found that introducing descriptors concerned with molecular flexibility can improve the linear correlation. The resulting model with four descriptors bears good statistical significance, n = 77, r = 0.82, q = 0.79, s = 0.45, F = 35.7. The actual predictive abilities of the QSAR model were validated through an external validation test set of 23 diverse compounds. The predictions for the tested compounds are as the same accuracy as the compounds of the training set and significantly better than those predicted by using the model reported. The good predictive ability suggests that the proposed model may be a good tool for fast screening of logPapp for compound libraries or large sets of new chemical entities via combinatorial chemistry synthesis.

Association of ethylene dibromide (EDB) with mature cranberry (Vaccinium macrocarpon) fruit.

Ethylene dibromide (EDB), a potential carcinogen, has been used in gasoline mixtures to avoid the accumulation of metallic lead in engines. Ethylene dibromide is present in the environment and in groundwater. Previous analysis has shown that EDB levels have reached up to 16 microg L-1 in the groundwater at two fuel spill plumes in the vicinity of the Massachusetts Military Reservation (MMR) Base and up to 1.69 microg L-1 in the Coonamessett and Quashnet Rivers in Cape Cod, MA (U.S. Air Force IRP, Fact Sheet #98-10, 1998). Groundwater and river water from this area are used to flood some local cranberry bogs for irrigation and harvesting of cranberry fruits. The potential sorption of EDB by cranberry fruits during harvest has caused concern but information regarding its occurrence is not available. In this study, low levels of EDB (0.04-0.15 microg kg-1) were found to be associated with cranberry fruits that were exposed to EDB at levels ranging from 3 to 12 microg L-1 at 10, 20, and 30 degrees C for up to 7 days. Rinsing EDB-exposed cranberry fruits twice with deionized water or once with 0.01 M NaCl solution reduced the amount of EDB associated with the cranberry fruits by 65-72% to a level of 0.02 microg kg-1. Therefore, the EDB most likely is associated with the water residue on the surface of the cranberry fruit rather than being absorbed into the flesh of the fruit during the EDB exposure.

[Mutation characteristic of STK].

OBJECTIVE: To identify the mutation characteristic of STK(11) gene in Chinese with Peutz-Jeghers syndrome(PJS) and establish the base of the gene diagnosis of PJS. METHODS: STK(11) germline mutation was analysed by DNA sequencing in 18 unrelation patients with PJS. RESULTS: Six novel mutations of STK (11) gene were detected in six unrelation patients. These mutations will lead to production of truncated protein. CONCLUSION: STK (11) gene mutation accounts for one third of the Chinese with PJS. The content of mutation includes single base substitution or deletion and one or two bases insertion. The mutations were widely found in different regions of the whole coding sequence, and 2/3 of those concentr ate in exon 1. Mutation frequency is 66.7% in the family suffering PJS in two or more generations, and 16.7% in the disseminated cases.

Tyrosine phosphorylation of the proto-oncoprotein Raf-1 is regulated by Raf-1 itself and the phosphatase Cdc25A.

There is a growing body of evidence demonstrating that Raf-1 is phosphorylated on tyrosines upon stimulation of a variety of receptors. Although detection of Raf-1 tyrosine phosphorylation has remained elusive, genetic analyses have demonstrated it to be important for Raf-1 activation. Here we report new findings which indicate that Raf-1 tyrosine phosphorylation is regulated in vivo. In both a mammalian and baculovirus expression system, a kinase-inactive allele of Raf-1 was found to be tyrosine phosphorylated at levels much greater than that of wild-type Raf-1. The level of tyrosine phosphate on Raf-1 was markedly increased upon treatment with phosphatase inhibitors either before or after cell lysis. Cdc25A was found to dephosphorylate Raf-1 on tyrosines that resulted in a significant decrease in Raf-1 kinase activity. In NIH 3T3 cells, coexpression of wild-type Raf-1 and phosphatase-inactive Cdc25A led to a marked increase in Raf-1 tyrosine phosphorylation in response to platelet-derived growth factor. These data suggest that the tyrosine phosphorylation of Raf-1 is regulated not only by itself but also by Cdc25A.

An ABA-binding protein with nucleic acid-binding property.

Maize cDNA expression library was screened using the antibodies against ABA-binding proteins (ABBP). From 200 000 separated phage plaques, only one cDNA clone coding for a polypeptide was obtained. It shows 60%-65% homology to the genes of nucleic acid-binding proteins in mammalia. However, about 120 cDNA clones coding maize 17s rRNA were unexpectedly obtained. In view of this, the ABBP purified by ABA affinity chromatography and the antibodies derived from the purified ABBP were examined. It is found that rRNA is indeed present in the ABBP. In addition, the obtained antibodies could recognize RNA as well as ABBP. It is suggested that the ABBP have nucleic acid-binding property and can form a complex with rRNA.

[Giant-cell tumor of bone: a review of diagnosis and treatment of 105 cases].

OBJECTIVE: To summarize the experience in the diagnosis and treatment of giant-cell tumor of bone, the selection of various operation methods and their effects were evaluated and discussed. METHODS: From 105 cases of giant-cell tumor of bone data were collected and reviewed. The clinical features were analyzed in association with radiological and histological characteristics. Prognosis was assessed by long-term follow-up. RESULTS: Among 105 cases, one had multiple giant-cell tumors and two had distant metastases. In some cases, the radiological and histological appearances didn't accord with the clinical findings. The recurrence rate was 36% in patients treated with curettage. Although no recurrence was seen in patients treated by local excision, the joint function could not be fully retained. CONCLUSION: Diagnosis should be made on the basis of integrated clinical, radiological and pathological manifestations. Early diagnosis and operation are important. The function of joint must be retained as much as possible. Pre-operative embolization of tumor of sacrum and pelvis is very helpful to ensure operative success and to minimize recurrence.

Erythropoietin activates Raf1 by an Shc-independent pathway in CTLL-EPO-R cells.

Stimulation of the erythropoietin receptor (EPO-R) or the interleukin-2 receptor (IL-2-R) by their respective ligands has been reported to activate tyrosine phosphorylation of the cytoplasmic protein, Shc. We have recently characterized a cell line, CTLL-EPO-R, that contains functional cell-surface receptors for both EPO and IL-2. Although stimulation with IL-2 or IL-15 resulted in the rapid, dose-dependent tyrosine phosphorylation of Shc, stimulation with EPO failed to activate Shc. EPO, IL-2, and IL-15 activated the tyrosine phosphorylation of the adaptor protein, Shp2, and the association of Shp2/Grb2/cytokine receptor complexes. In addition, EPO, IL-2, and IL-15 activated Raf1 and ERK2, demonstrating that the Raf1/MEK/MAP kinase pathway was activated. These results indicate that multiple biochemical pathways are capable of conferring a mitogenic signal in CTLL-EPO-R. EPO can activate the Raf1/MEK/MAP kinase pathway via Shc-dependent or Shc-independent pathways, and Shc activation is not required for EPO-dependent cell growth in CTLL-EPO-R.

The cytokine-activated tyrosine kinase JAK2 activates Raf-1 in a p21ras-dependent manner.

JAK2, a member of the Janus kinase superfamily was found to interact functionally with Raf-1, a central component of the ras/mitogen-activated protein kinase signal transduction pathway. Interferon-gamma and several other cytokines that are known to activate JAK2 kinase were also found to stimulate Raf-1 kinase activity toward MEK-1 in mammalian cells. In the baculovirus coexpression system, Raf-1 was activated by JAK2 in the presence of p21ras. Under these conditions, a ternary complex of p21ras, JAK2, and Raf-1 was observed. In contrast, in the absence of p21ras, coexpression of JAK2 and Raf-1 resulted in an overall decrease in the Raf-1 kinase activity. In addition, JAK2 phosphorylated Raf-1 at sites different from those phosphorylated by pp60v-src. In mammalian cells treated with either erythropoietin or interferon-gamma, a small fraction of Raf-1 coimmunoprecipitated with JAK2 in lysates of cells in which JAK2 was activated as judged by its state of tyrosine phosphorylation. Taken together, these data suggest that JAK2 and p21ras cooperate to activate Raf-1.

Role of protein kinase A and the serine-rich region of herpes simplex virus type 1 ICP4 in viral replication.

Efficient expression of herpes simplex virus genes requires the synthesis of functional ICP4, a nuclear phosphoprotein that contains a prominent serine-rich region between amino acids 142 and 210. Residues in this region not only are potential sites for phosphorylation but also are involved in the functions of ICP4. By comparing the growth of a virus in which this region is deleted (d8-10) with wild-type virus (KOS) in PC12 cells or PC12 cells that are deficient in cyclic AMP-dependent protein kinase (PKA), two observations were made: (i) the growth of wild-type virus was impaired by 1 to 2 orders of magnitude in the PKA-deficient cells, indicating the involvement of PKA in the growth cycle of herpes simplex virus type 1, and (ii) while the growth of d8-10 was impaired by almost 2 orders of magnitude in wild-type cells, it was not further impaired (as was that of wild-type virus) in PKA-deficient cells, implicating the region deleted in d8-10 as a possible target for cellular PKA. In trigeminal'ganglia of mice, the d8-10 mutant virus grew poorly; however, it established latency in nearly 90% of ganglia tested. Studies of the phosphorylation of wild-type and d8-10 ICP4 proteins revealed that the serine-rich region is a major determinant for phosphorylation of ICP4 in vivo and that the phosphorylation state could change as a function of the PKA activity. Consistent with this observation, the serine-rich region of ICP4 was shown to be a target for PKA in vitro. While intact ICP4 was readily phosphorylated by ICP4 in vitro, the d8-10 mutant ICP4 was not. Moreover, a synthethic peptide representing a sequence in the serine tract that is predicted to be a substrate for PKA was phosphorylated by PKA in vitro, having a Km within the physiological range. These data suggest that PKA plays a role in viral growth through phosphorylation of one or more sites on the ICP4 molecule.

Analysis of phosphorylation sites of herpes simplex virus type 1 ICP4.

The herpes simplex virus ICP4 protein is required for induction of early and late viral gene transcription as well as for repression of expression of its own gene and several other viral genes. Several electrophoretic forms of ICP4 have been observed, and phosphorylation is thought to contribute to this heterogeneity and possibly to the multiple functions of ICP4. To define the complexity of the site(s) of phosphorylation of ICP4 and to initiate mapping of this site(s), we have performed two-dimensional phosphopeptide mapping of wild-type and mutant forms of ICP4 labeled in infected cells or in vitro. Wild-type ICP4 labeled in infected cells shows a complex pattern of phosphopeptides, and smaller mutant forms of ICP4 show progressively fewer phosphopeptides, arguing that multiple sites on ICP4 are phosphorylated. The serine-rich region of ICP4, residues 175 to 198, was shown to be a site for phosphorylation. Furthermore, the serine-rich region itself or the phosphorylation of this region increases phosphorylation of all phosphopeptides. A mutant ICP4 molecule lacking the serine-rich region showed low levels of phosphorylation by protein kinase A or protein kinase C in vitro. These results suggest that there may be a sequential phosphorylation of ICP4, with phosphorylation of the serine-rich region stimulating phosphorylation of the rest of the molecule. In addition, purified ICP4 showed an associated kinase activity or an autophosphorylation activity with properties different from those of protein kinase A or protein kinase C.

Interaction of the protein kinase Raf-1 with 14-3-3 proteins.

Members of a family of highly conserved proteins, termed 14-3-3 proteins, were found by several experimental approaches to associate with Raf-1, a central component of a key signal transduction pathway. Optimal complex formation required the amino-terminal regulatory domain of Raf-1. The association of 14-3-3 proteins and Raf-1 was not substantially affected by the activation state of Raf.

[Studies of cockroachcidal toxin].

A cockroachcidal bacterial isolate CW-W-90-3 was selected by egg yolk agar plate. The isolate produced phospholipase C (PLC) which was pathogenic to the nymph of cockroach. The conditions for production of high level PLC indicated that using LB medium supplement Tween-80 or minimal medium could effectively increase the activity of PLC. The optical phase for production of PLC was in the period of 12-18 hours and below pH8.0. The activity of PLC was reduced along with the culture time until 48 hours. The PLC was resistant to heat. The partially purified PLC from the culture supernatant was assayed by using cockroach nymphes and produced 71.74% mortality.