[Comparative study of gap balancing and measured resection technique in patients receiving staged bilateral total knee arthroplasty].

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

Action of the insecticide cyfluthrin on Ca2+ signal transduction and cytotoxicity in human osteosarcoma cells.

Cyfluthrin is a pyrethroid insecticide and common household pesticide. The effect of cyfluthrin on Ca2+-related physiology in human osteosarcoma is unclear. This study investigated the effect of cyfluthrin on cytosolic-free Ca2+ concentrations ([Ca2+]i) and viability in MG63 human osteosarcoma cells. Cyfluthrin concentration-dependently induced [Ca2+]i rises. Cyfluthrin-induced Ca2+ entry was confirmed by the Mn2+-induced quench of fura-2 fluorescence. Cyfluthrin at concentrations of 10-100 µM induced [Ca2+]i rises. Ca2+ removal reduced the signal by approximately 50%. Cyfluthrin (100 µM) induced Mn2+ influx suggesting Ca2+ entry. Cyfluthrin-induced Ca2+ entry was inhibited 50% by protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate) and inhibitor (GF109203X) and also by three inhibitors of store-operated Ca2+ channels: nifedipine, econazole, and SKF96365. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (TG) completely inhibited cyfluthrin-evoked [Ca2+]i rises. Conversely, treatment with cyfluthrin abolished TG-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with 1-[6-[((17ß)-3-methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dion abolished cyfluthrin-induced [Ca2+]i rises. Cyfluthrin at 25-65 µM decreased cell viability, which was not reversed by pretreatment with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester. Together, in MG63 cells, cyfluthrin induced [Ca2+]i rises by evoking PLC-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via PKC-sensitive store-operated Ca2+ entry. Cyfluthrin also caused Ca2+-independent cell death.

Ca2+ movement and cytotoxicity induced by the pyrethroid pesticide bifenthrin in human prostate cancer cells.

Bifenthrin, a commonly used pyrethroid pesticide, evokes various toxicological effects in different models. However, the effect of bifenthrin on cytosolic-free Ca2+ level ([Ca2+]i) and cytotoxicity in human prostate cancer cells is unclear. This study examined whether bifenthrin altered Ca2+ homeostasis and cell viability in PC3 human prostate cancer cells. [Ca2+]i in suspended cells were measured using the fluorescent Ca2+-sensitive dye fura-2. Cell viability was examined by 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] water soluble tetrazolium-1 assay. Bifenthrin (100-400 µM) concentration-dependently induced [Ca2+]i rises. Ca2+ removal reduced the signal by approximately 30%. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) abolished bifenthrin-evoked [Ca2+]i rises. Conversely, treatment with bifenthrin abolished BHQ-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 significantly inhibited bifenthrin-induced [Ca2+]i rises. Mn2+ has been shown to enter cells through similar mechanisms as Ca2+ but quenches fura-2 fluorescence at all excitation wavelengths. Bifenthrin (400 µM)-induced Mn2+ influx implicates that Ca2+ entry occurred. Bifenthrin-induced Ca2+ entry was inhibited by 30% by protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate) and inhibitor (GF109203X) and three inhibitors of store-operated Ca2+ channels: nifedipine, econazole, and SKF96365. Bifenthrin at 175-275 µM decreased cell viability, which was not reversed by pretreatment with the Ca2+ chelator 1,2-bis(2-aminophenoxy) ethane-N,N,N',N'-tetra acetic acid-acetoxymethyl ester. Together, in PC3 cells, bifenthrin-induced [Ca2+]i rises by evoking PLC-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via PKC-sensitive store-operated Ca2+ entry. Bifenthrin also caused Ca2+-independent cell death.

Phorbol ester-potentiated liposomal transfection to monocytic PLB-985 cells.

In order to improve transient gene transfer into PLB-985 cells, we treated cells with 12-tetradecanoylphorbol 13-acetate (TPA) 4 h before transfection, and increased by 540-fold the reporter activity of the firefly luciferase gene transfected by TFL-01, a cationic liposome. Dioctanolyglycerol added before TPA addition inhibited the TPA-dependent increase in activity, suggesting it to be a TPA competitor for PKC binding. H7, staurosporine, GO 6976, and H8, but not GF 109203X and forskolin, inhibited the TPA-dependent increase in reporter activity when added 8 h after TFL-01/gene addition. Forskolin and GF 109203X also inhibited the increase when added before TPA. Therefore, for the potentiation of transfection by the TPA/TFL-01 method, conventional PKC activity with significant but low protein kinase A (PKA) activity are first required, and then a novel PKC activity with significant PKA activity. TPA enhanced the uptake of FITC-labeled phosphorothioated oligonucleotides and their prolonged maintenance by cells, suggesting increased TFL-01-assisted plasmid uptake and its stabilization in TPA-treated PLB-985 cells. This method was used successfully for the sensitive analysis of the promoter function of the gp91(phox) gene, implying the method to be generally useful for promoter analyses of various genes expressed in differentiated human monocytic cells.

Short- and long-term effect of interferon therapy for chronic hepatitis C.

AIM: To study the short and long-term effect of interferon therapy for chronic hepatitis C. METHODS: Sixty-eight patients with chronic hepatitis C were treated with interferon (3 × 10(6) IU, im/2 d, for a course of three months) with 1 to 5 courses of treatment and followed for 1.5 to 3 years after the therapy. RESULTS: According to antiviral effect of interferon, 76.5% (52/68) of the cases had a complete response by the end of the first therapy course, while 20.6% (14/68) and 2.9% (2/68) had a partial response or non-response. Over a half of the patients with a complete response (27/52, 51.9%) relapsed within 6 to 10 mo after the first course. Of the original cohort, nineteen patients received two courses of therapy, while one patient received three and another three received five courses of therapy. The follow-up for these patients was between 1.5 to 3 years, at which time 29 (42.7%) of the patients sustained a complete response, with four of them having HCV RNA positive serum, while the others had either a partial (37/68, 2.9%) or non-response. CONCLUSION: Interferon therapy had a high short-term complete response but a low long-term complete response in the treatment of chronic hepatitis C.

A study of the binding ability of pre-S1 and S2 proteins of hepatitis B virus to human serum albumin.

To assess the relationship between pre-S proteins of HBV and polymerized human serum albumin (PHSA), a labeled avidin biotin ELISA was used to detect pre-S1, S2 and PHSA receptor (PHSAR) activity. PHSAR activity was only present in the samples positive for pre-S1 and/or S2, but not in the samples positive only for HBsAg. Pre-S1, S2 and PHSAR could in some degree be blocked by preincubating serum with PHSA, and the blocking efficiency of PHSA with respect to pre-S2 and PHSAR was similar, suggesting that pre-S2 is the dominant site for binding PHSA in vitro. We also found that PHSAR activity was detectable in 2 cases positive only for pre-S1, but not pre-S2. Furthermore, PHSA could selectively block pre-S1 and PHSAR activity in 2 cases negative for pre-S2, revealing that pre-S1 also possesses binding ability to PHSA, at least in a small number of cases. Using sandwich ELISA, we demonstrated the existence of complexes of HBV envelope proteins and human serum albumin (HSA) in some HBV infected serum samples. The possible significance of these complexes is discussed.

Hepatitis D virus infection in liver tissues of patients with hepatitis B in China.

2,346 liver samples from 17 cities of China for intrahepatic hepatitis D antigen (HDAg) were studied by direct enzyme-labelled technique. HDAg was detected in 167 out of 1,764 samples of HBsAg positive individuals making a detection rate of 9.47%. Hepatitis D virus (HDV) infection existed in all the examined districts with no significant difference in the HDAg detection rate. It was found that the intrahepatic HDAg detection rate was related to the pathologic type of the liver disease. The HDAg detection rate in chronic liver diseases and severe hepatitis was higher than in other liver diseases. It suggests that HDV infection is associated with the progression and chronicity of the liver disease. Studies on the relationship between HDV infection and HBV replication showed that HBV replication might be suppressed by HDV infection. Both HDV and HBV, however, could replicate in the same hepatocyte simultaneously.

Antibodies to hepatitis C virus and hepatitis C virus RNA in Chinese blood donors determined by ELISA, recombinant immunoblot assay and polymerase chain reaction.

Antibodies to hepatitis C virus (anti-HCV) were determined in Chinese blood donors from the city of Wuhan by ELISA screening tests and confirmatory recombinant immunoblot assay (RIBA). 281 and 222 sera were sampled under similar conditions in 1989 and 1990, respectively. The first collection of sera was sent to Sweden in lyophilized form, the second directly as fresh, unfrozen sera. A high proportion (22%) of the lyophilized sera were positive in the screening assay but only 6 (2.10%) were positive by RIBA with antibodies against both the C100-3 and 5-1-1 peptides. HCV RNA could be detected by polymerase chain reaction (PCR) analysis in 3 of the 6 sera and in one reacting with C100-3 only. In the second material of fresh sera only 3 were positive in the screening anti-HCV ELISA, but none was RIBA or PCR positive. Thus, the overall prevalence of anti-HCV among the 503 Chinese blood donors as identified by RIBA was 1.2%, and that of HCV RNA by PCR was 0.8%. The confirmed antibody prevalence is higher than that reported in the western literature.

Hepatitis B virus DNA detected by PCR in sera and liver tissues of Chinese patients with chronic liver diseases.

To investigate the HBV infection and its replication in Chinese patients with chronic liver disease, polymerase chain reaction (PCR) was employed to detect hepatitis B virus DNA in sera of 410 patients with chronic liver disease and liver samples from 188 patients. The HBV DNA detectability in all serum samples was 58%. Among them 100% HBeAg-positive and 58% anti-HBe cases were HBV DNA detectable respectively. However, HBV DNA was also found in 23% HBsAg-negative/anti-HBc positive chronic cases. Furthermore, 30% anti-HBs positive chronic cases who had neutralizing antibody against HBV infection, continued to contain HBV DNA. Our findings indicate that HBV infection and its replication are dominant cause of chronic liver disease and some HBV variants may escape from the protective antibody to induce chronic liver disease, even anti-HBs antibody circulated.

Relationship between the characteristics of immunopathological expression of hepatitis C virus antigen and hepatocytic injury.

Biopsied liver tissues from 352 cases were tested for hepatitis C virus (HCVAg) with improved PAP immunohistologic chemical method. Furthermore, corresponding seroantibody to hepatitis C virus was also tested. The total HCVAg positive rate was 9.1%. The HCVAg positive rate in chronic persistent hepatitis (CPH) was 5%. The HCVAg positive rate in chronic active hepatitis (CAH) was 11.2%. The HCVAg positive rate raised gradually along with the severity of hepatocytic injury. HCVAg may be seen in necrotic liver cells exfoliating into the liver sinus, indicating a close relationship between HCVAg and hepatocytic injury. Expression of HCVAg was mostly of the nucleus type in CPH cases and was mostly of the plasma type in CAH cases. The periphery of nucleus type-expressed positive cells generally had no marked inflammatory cell infiltration. The periphery of plasma type-expressed positive cells had a certain amount of inflammatory cell infiltration. Along with the severity of hepatocytic injury, HCVAg expressed itself in a positive correlation according to the nucleus and plasma types. The HCVAg positive cells were located mostly in the lobular peripheral band and rarely located in the venoperipheral band. It was possible that this had some relation with the lobular microcirculation of blood and blood supply. In this study, there was no obvious correlation between the HCVAg positive rate in hepatic tissues and the anti-HCV positive rate in sera. Neither the patients with HCVAg positive liver tissues nor the patients with seropositive anti-HCV had any history of blood transfusion and the use of blood products.(ABSTRACT TRUNCATED AT 250 WORDS)

Hepatocyte necrosis associated with HBV marker expression and T cytotoxic cells in situ in chronic hepatitis B.

To explore the role of HBV antigen expression and monocyte infiltrate in situ in chronic hepatitis, HBV markers and compositions and number of monocytes in the liver of chronic hepatitis B were immunohistochemically located and identified, and correlated with hepatocyte necrosis. It was found that hepatic necrosis frequently took place in the centre or boundary of membranous HBsAg and/or HBcAg expression and the majority of monocytes accumulated in the necrotic areas showed CD8+ cell token in HBeAg-positive stage of chronic active hepatitis. On the other hand, with HBeAg positivity converted into HBeAg-negativity or anti-HBe-positivity, HBV antigen expression significantly declined, CD4+ cells remarkably increased as compared to HBeAg-positive status. These findings suggest that there may be distinct pathogenesis of hepatic necrosis in different stages of chronic HBV infection.

Analysis of intrahepatic HBVDNA in serologic HBV markers-negative or HBsAg-negative patients with chronic hepatitis.

To explore HBV infection status in a group of serologic HBV markers-negative or HBsAg-negative patients with chronic hepatitis, in situ hybridization to hepatocellular HBVDNA was carried out in combination with detection of HBsAg and HBcAg in the liver tissues. It was found that prevalence of intrahepatic HBVDNA, HBsAg and HBcAg was 43%, 39% and 17% respectively, and 15 out of 23 cases studied were sure to bear one or more positive marker(s) of intrahepatic HBVDNA, HBsAg and HBcAg. These findings suggest that more than half of the patients with chronic hepatitis were still undergoing HBV infection, despite serologic HBV markers or HBsAg negative. Furthermore, we found that hepatocytes containing HBVDNA or surface or core antigen were often close to hepatic necrosis foci, indicating that HBV replication and its antigen(s) expression in hepatocytes could account for chronic, active and necrotic inflammation occurring in the liver of the patients mentioned above.

Molecular cloning and characterization of major histocompatibility complex class I cDNAs from woodchuck (Marmota monax).

The full-length cDNAs of woodchuck major histocompatibility complex (MHC) class I (MhcMamo-I or Mamo-I) genes were cloned by using cellular mRNA isolated from the peripheral blood mononuclear cells and liver tissues of woodchucks. DNA sequence analysis of Mamo-I cDNAs revealed that the coding regions of Mamo-I genes were about 1,080 bp long, encoding 359 amino acid residues. The deduced amino acid sequences of Mamo-I showed structural features like leader, alpha1, alpha2, alpha3, transmembrane and cytoplasmic domains, similar to their homologues in human and other mammals. Analysis of five full-length clones from unrelated woodchucks indicated a polymorphism within the alpha1 and alpha2 domains of Mamo-I heavy chain and a high conservation within the alpha3 and the transmembrane/cytoplasmic domains. Amino acid residues of the alpha2 and alpha3 domains that are supposed to be involved in the binding of MHC class I to CD8 molecule, were largely conserved in Mamo-I genes. Phylogenetic comparison of MHC class I genes of woodchuck and other mammals indicated a close evolutionary relationship between woodchuck and squirrel MHC class I. We tentatively named this region the locus A of Mamo-I genes (Mamo-A). Sequence analysis of 101 clones of alpha1 and alpha2 regions derived from 14 woodchucks revealed that at least 14 different alleles within Mamo-A exist. Among these 14 alleles identified so far, Mamo-A*01 and Mamo-A*09 were of the highest frequency of about 21.5% and 14.5%, respectively. Our results indicate that Mamo-I genes are of a similar molecular structure to those of human and other mammals.

Intrahepatic expression of pre-S proteins of hepatitis B virus and its possible relation to liver cell necrosis.

To assess the significance of intrahepatic expression of pre-S1 and S2 proteins of hepatitis B virus (HBV) in patients with HBV infection, an indirect immunoperoxidase technique employed monoclonal antibodies to pre-S proteins was used to detect pre-S1 and -S2 proteins in 80 liver specimens. The frequency of pre-S1 and -S2 proteins was 61.3% and 51.3%, respectively, and the co-expression of pre-S and HBsAg occurred in most specimens. The preferential expression of pre-S1 and -S2 in HBcAg-positive specimens suggests that pre-S proteins are associated with HBV replication. Membranous expression of both pre-S1 and -S2 is associated with inflammatory activity and liver cell necrosis. Furthermore, our results show that T cells, not NK or B cells, were the predominantly infiltrating cells in necrotic foci with pre-S expression. Almost all of these T cells may express HLA-DR antigen simultaneously; therefore, they are activated. In conjunction with these data, we conclude that, as the essential components of HBV envelope proteins, pre-S proteins may play an important role in resulting in liver cell necrosis.