[Screening results and genetic analysis of neonatal tetrahydrobiopterin deficiency in Hainan Province from 2007 to 2019].

A total of 1 295 516 dried blood spots were collected from newborns in Hainan Province from 2007 to 2019 who participated in the screening of neonatal diseases, and 43 cases of hyperphenylalaninemia were diagnosed. Among the 43 cases, 8 cases were confirmed to have tetrahydrobiopterin deficiency (4 males and 4 females). The incidence of tetrahydrobiopterin deficiency among newborns in Hainan Province was 6.2/1 million. Six mutations in the PTS gene were detected among 7 cases; the mutations were as follows: c.317C>T, c.286G>A, c.259C>T, c.155A>G, c.84+291A>G and c.83+1777T>G. A homozygous mutation at c.41T>C site of QDPR gene was detected in one case. Overall, it's found that the incidence of tetrahydrobiopterin deficiency in newborn populations in Hainan Province is low, and PTS gene mutations account for the largest proportion of cases of tetrahydrobiopterin deficiency within the study population.

Cone Beam Computed Tomography in observing the presence and location of canalis sinuosus.

OBJECTIVE: Cone Beam Computed Tomography (CBCT) was used to observe and describe the distribution of canalis sinuosus (CS) in the Chinese population and the location of CS in the maxillary alveolar bone, so as to help oral surgeons evaluate the intraoperative risk and prognosis before maxillary surgery and reduce the complications caused by the injury of this structure in anterior surgery. PATIENTS AND METHODS: CBCT images of 600 patients admitted from 2021 to 2022 were collected to observe the anatomical structure of CS in the maxillary region. The following parameters were recorded: age, sex, number of CS, left and right distribution of CS, CS diameter, and location. Statistical analysis was performed on all of the collected data. RESULTS: The discovery rate of CS in this study was 59.75%, and it is commonly found in the lateral incisor area (64.82%). No significant difference can be found in the presence and number of CS in different gender and age groups (p>0.05). CONCLUSIONS: The use of high-resolution CBCT before implantation is of irreplaceable significance in the diagnosis and analysis of CS, which is conducive to reducing implantation complications and failure rate. The incidence of CS was independent of age or sex, while the location of CS was statistically significant.

Clinical analysis of head and neck cancer cases in south-west China 1953 - 2002.

The occurrence of head and neck cancers varies worldwide. This study retrospectively analysed the incidence and types of 8646 cases of head and neck cancers in south-west China that were treated at the West China Stomatology Hospital of Sichuan University between 1953 and 2002. Overall mean patient age was 50.3 years and the overall male:female ratio was 2.38:1; mean age increased and the male:female ratio decreased over the study period. Peak incidence occurred between the ages of 40 and 60 years. Primary tumours most frequently developed in the tongue, followed by the bucca and gingiva. Histologically, squamous cell carcinomas were most frequently recorded. The parotid gland and palate were the most common locations for salivary gland tumours. Over the study period the incidence of head and neck cancers increased with time and the rate of increase was greater in females than males. The frequency of histological types and topography were similar to previous reports.

Immunization against hepatitis B virus by mucosal administration of antigen-antibody complexes.

Antigen-antibody complexes have been shown to enhance immune responses against several antigens given by parenteral immunization. Herein, we have evaluated the potential of administering such immunostimulatory complexes by a mucosal route. Hepatitis B surface antigen (HBsAg) complexed with antibodies against HBsAg (anti-HBs) (HBsAg/Ab) was administered to BALB/c mice by intranasal inhalation. HBsAg by itself did not induce immune responses, whereas with HBsAg/Ab complexes, both systemic and mucosal immune responses were observed and these could be modulated by adjuvants. With HBsAg/Ab (1 or 10 microg), anti-HBs antibodies induced were predominantly of the IgG1 isotype (Th2-like). In contrast, anti-HBs induced by HBsAg/Ab plus cholera toxin (CT) or oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs (CpG) (1 microg each) were predominantly IgG2a (Th1-like). Results from this study indicate that HBsAg/Ab complexes can induce strong humoral immune responses when delivered by a noninvasive route, whether used alone or in combination with other mucosal adjuvants.

Comparing the immunogenicity of hepatitis B virus S gene variants by DNA immunization.

DNA immunization was used to compare the immunogenicity of hepatitis B virus S gene variants. Four recombinant plasmid DNAs containing the full-length virus genome with different S gene inserts were used to immunize BALB/c and C57/BL/6 mice. These inserts were cloned from 129L (residue 129, glutamine to leucine), 129H (residue 129, glutamine to histidine) 145R (residue 145, glycine to arginine) variants and the wild-type virus. The titer of hepatitis B virus core antibodies (anti-HBc) in immunized mice was used as the control for the efficiency of DNA immunization. Serum hepatitis B surface antibody (anti-HBs) titer and cytokines induced in splenocytes stimulated with hepatitis B surface antigen (HBsAg) were monitored as specific immune responses induced by different plasmid DNAs. 129L DNA induced significantly lower anti-HBs antibodies (IgG, IgG1, and IgG2a) and less interferon-gamma, compared to those in mice immunized with the 129H variant and the wild-type HBV DNA (p < 0.05). Computer modeling showed that a change from glutamine to leucine at 129 residue led to higher hydrophobicity and could result in decreased immunogenicity. Results indicate that DNA immunization can be used to compare the humoral and cellular immunogenicity among different HBV S variants.

Effect of plasmid DNA on immunogenicity of HBsAg-anti-HBs complex.

Hepatitis B surface antigen (HBsAg) complexed with anti-HBs is more immunogenic than HBsAg alone in mice. This complex is usually used with alum as an adjuvant, which can enhance humoral response but inhibits cell-mediated immune responses. To improve the immunogenicity of HBsAg-anti-HBs, we immunized mice with a combination of this immunogenic complex and pCMVHBs, a plasmid encoding HBsAg, or the vector pCMV. Both plasmids enhanced the anti-HBs response induced by the immunogenic complex. We found 20 microg of plasmid or vector enhanced the anti-HBs response in all mice, whereas 1 microg was less effective. Splenocytes from different immunized groups were stimulated with HBsAg in vitro, and the highest level of IL-2 detected in the supernatant was found in mice immunized with HBsAg-anti-HBs plus pCMVHBs. A plasmid (pcDNA3c191) encoding core protein of hepatitis C virus (HCV) was used as an adjuvant to the immunogenic complex. A preliminary result showed that pcDNA3c191 not only enhanced the immunogenicity of HBsAg-anti-HBs, but also induced anti-HCV core antibodies. Immunization using a plasmid DNA encoding one viral antigen in combination with antigen and antibody complex of another microbial origin could be a new approach to the development of multivalent vaccines.

Antigen-antibody complex as therapeutic vaccine for viral hepatitis B.

In a previous study, hepatitis B surface antigen (HBsAg) complexed to human anti-HBs immunoglobulins (HBIG) in excess of HBsAg was used as therapeutic vaccine to treat chronic hepatitis B patients and promising results were obtained. To study the mechanisms of this approach, mice were immunized with HBsAg or IC (immunogenic complex, i.e. HBsAg complexed with mouse polyclonal anti-HBs). Studies indicate that IC induced enhanced immune responses by increasing uptake of HBsAg through Fc receptors on antigen presenting cells and modulated HBsAg processing and presentation. This modulation led to stimulation of T cell responses, and increased production of IL-2 and IFN-gamma. Assay for antibody subclasses showed that higher ratio of IgG 2a was observed in the IC immunized group, which correlated with the production of lymphokine pattern. When alum was used as the adjuvant, though antibody response was enhanced, production of cytokines decreased. When DNA from a recombinant plasmid was added to IC as an adjuvant, the titer of anti-HBs was significantly higher than those in mice immunized only with the DNA or the IC. Since DNA immunization can induce both cellular and humoral immune responses, combined immunization using IC and DNA might serve as another type of therapeutic vaccine for viral hepatitis B.

Exploiting new potential targets for anti-hepatitis B virus drugs.

Based on the recent studies of HBV strains with different replication efficiency, several new potential targets for anti-HBV replication have been presented. These include the viral and cellular regulatory factors associated with HBV replication and the process for encapsidation of viral genome and budding into endoplasmic reticulum (ER). A putative regulatory domain has been reported at the carboxyl-end of reverse transcriptase (RT) and when serine is substituted for proline at residue 652 of RT, replication efficiency of HBV is decreased. Substitution of proline for threonine at the 2798 nucleotide of the terminal protein (TP) gene, renders the mutant completely replication deficient. Expression of TP blocks the interferon (IFN) pathway and inhibits the responsive state of cells to interferons ( IFN) alpha and gamma. Interference of HBV capsid assembly drastically affects the encapsidation of viral genome, a crucial process for reverse transcription and viral DNA synthesis. Small molecules (bis-ANS) have been reported to act as a "wedge" to misdirect the polymerization of capsid, resulting in inhibition of virus replication. Another new group of compounds (HAP) has been shown to inhibit virus replication and also inhibit the assembly of viral capsid (core particle). Finally the capsids containing HBV genome are enveloped by budding into endoplasmic reticulum and release from virus infected cells, and this morphogenesis and secretion of HBV is dependent on glucosidases in the ER of host cells. Competitive inhibition of these glucosidases has been suggested as strategy against HBV replication.

Detection of HPV in Japanese and Chinese oral carcinomas by in situ PCR.

Human papillomavirus (HPV) is established as the cause of almost 100% of cervical carcinomas. However, the association of HPV with oral squamous cell carcinomas (SCCs) is less well understood. We examined the prevalence of HPV in oral SCCs in samples of Japanese and Chinese populations. Using in situ polymerase chain reaction (PCR) analysis (MY09 and MY11 consensus primers), HPV was detected in the nucleus of epithelia and tumor cells in oral lesions. Analysis revealed the specific presence of HPV DNA in all cases of SCC in our Japanese (10/10) and Chinese (10/10) population samples. These results suggest that HPV infection could be one of several risk factors contributing to oral SCC in Japanese and Chinese.

Cloning and Expression of Chinese Duck Interferon-gamma Gene.

The efficacy of cytokine therapy has been demonstrated in several viral diseases. Interferon-gamma is a cytokine that has potent antiviral property and immunomodulatory activity. To investigate the role of IFN-gamma in viral clearance during natural infection and to define the antiviral mechanism, DHBV-infected ducks was used as an animal model. To clone, express, and develop the method of quantifying DuIFN-gamma gene transcription and expression, DuIFN -gamma cDNA was amplified by RT-PCR from PHA stimulated duck PBMC. Recombinant plasmid expressing DuIFN-gamma was used to transfect COS-7, and the cell culture supernatant was analyzed by CPE inhibitory assay and MTT methods to determine the antiviral titer of IFN-gamma. The GST-DuIFN-gamma fusion protein was expressed in E.coli and purified using the GST sepharose 4B. Results indicated that the supernatant collected from COS-7 cells transfected with DuIFN-gamma cDNA was able to prevent duck fibroblasts from VSV induced CPE in a dose dependent manner. An anti-DuIFN-gamma antibody neutralized this antiviral activity.

Enhanced immunogenicity in mice with hepatitis B vaccine complexed to human hepatitis B immunoglobulin.

Purified human hepatitis B immunoglobulin (HBIG) was complexed to plasma derived hepatitis B vaccine (HBVac) at different concentrations and used to immunize Balb/c mice. An enhanced humoral immune response was observed when HBVac was complexed to HBIG in excess of antigen, compared to that immunized with the vaccine alone. Proliferation of splenic lymphocytes was detected when mice were immunized with HBIG complexed to HBVac (0.2-1 microgram), whereas in mice immunized only with HBVac at one microgram, no lymphocyte proliferation was observed. The enhanced immunogenicity of HBIG: HBVac is T cell dependent. The importance of using critical ratio of HBIG and HBVac is indicated, and future application of this complex for vaccination of low- or nonresponders to the present HBVac, as well as for treatment of chronic hepatitis B patients is discussed.

Elimination of immune tolerance to hepatitis virus in an animal model.

Four-day old ducklings were infected with duck hepatitis B virus to simulate perinatal transmission of hepatitis B virus. Immune tolerance was characterized as persistent viremia, antigenemia, without detection of anti-DHBs or anti-DHBc. A synthetic peptide, P125-146, mimicking one of the epitopes of the native DHBV Pre-S protein was used to cross-link to tetanus toxoid or phytohemagglutinin. These 'novel' antigens were used to immunize immune tolerant ducks, aimed at bypassing T cell tolerance. After 5 injections, though no anti-DHBV Pre-S was detected, around 50% of the immunized ducks showed seroconversion to DHBV DNA negative.

Location of myocardial infarction and its surface cardioelectric representation. An experimental study.

In order to explore the relationship between the different location of myocardial infarction (MI) and reflections of pathognomonic Q potentials on the body surface, an experimental study was carried out in 28 dogs. The results were (1) in 27 of a total 28 dogs after MI formation, abnormal Q maps appeared; (2) right ventricular MI, posterior wall MI and some lateral and apical MI are prone to miss abnormal Q potentials in the conventional 12-lead ECG; and (3) the MI location and relative size diagnosed through electrocardiographic peak mapping (EPM) correlated to acute MI in dog ventricles fairly well. The unexpected Q map reflections of right ventricular MI provide new clues for improving our automatic diagnosis system after further correlative studies. The results strongly support the feasibility of the application of EPM in clinical diagnosis.

Recombinant vaccinia virus expressing Pre-S/S protein of duck hepatitis B virus and its preliminary use for treatment of persistent infection.

The envelope (Pre-S/S) gene of duck hepatitis B virus (DHBV) was amplified by polymerase chain reaction (PCR) and cloned into plasmid pGJP5, under the control of vaccinia virus promoter P(7.5). By recombination in cell culture, and screened in human TK- 143 cells in the presence of 5-bromouracil deoxyriboside (5-BUdR), a recombinant vaccinia virus, bearing the envelope gene of DHBV (pGDHBV-5) which could replicate in cell cultures was constructed. DHBV surface antigen (DHBsAg) was detected in pGDHBV-5-infected cell lysate by dot enzyme immunoassay (EIA). After multiple-site intradermal injections of pGDHBV-5, DHBsAg could be detected in the serum of immunized adult ducks. This indicated that the recombinant virus replicated and expresed DHBsAg in ducks. The recombinant virus was used as a therapeutic vaccine to immunize persistently DHBV-infected ducks. After immunization, a transient significant decrease of serum DHBsAg was observed.

[Study on cisplatin albumin microspheres for neck external artery embolization].

In this paper the technique of emulsion chemical-crosslinking was used to prepare cisplatin albumin microsphere for jaw squamous cancer by neck external artery embolization. It was yellow powder with yield 80 +/- 5%, mean size 56.3 microns, cisplatin concentration 14.02-14.20%, loading rate 97.08-97.95%. The release characteristics in vitro, sterilization, stability and recipe of disperse solvent of cisplatin albumin microsphere were investigated. Animal test showed that cisplatin albumin microsphere may plug the branches of neck external artery well and remain in local tissue.

[Studies on the hepatitis B virus in cell culture: I. Expression of cloned HBV DNA in the HEP G2 cell line].

To study the interactions between HBV and host cells, cloned adr subtype HBV DNA dimer was used to transfect Hep G2 cell line. Five days after transfection, HBeAg could be detected in the supernatant and reached its peak on the 11th day. It was still positive on the 25th day. In contrast, HBsAg was only positive on the 8th day, and gradually increased until the 25th day when the amount of HBsAg was still rising. Supernatant of HBeAg positive samples was collected and after ultracentrifugation, was checked by electron microscopy. HBV like particles of 42 nm in diameter were detected. The prospect of employing this transient cell expression system for studies of HBV replication, inhibitory effects of drugs and immunological factors are discussed.

[Persistent microbial infection and therapeutic vaccine].

Microbial persistent infection usually occurs due to invasion of the immune system, microbial genomic mutation or integration of microbial DNA with host chromosomal DNA. Defective host immune responses or immune tolerance are also related to persistent infection. Therapeutic vaccine is aimed at inducing specific humoral and cellular immune responses to terminate microbial persistent infection or to prevent progression of diseases. Different categories of therapeutic vaccines have been presented and the prospect of research as well as the development of novel effective therapeutic vaccines are discussed.

ATP-mediated protein kinase B Akt/mammalian target of rapamycin mTOR/p70 ribosomal S6 protein p70S6 kinase signaling pathway activation promotes improvement of locomotor function after spinal cord injury in rats.

The protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/p70 ribosomal S6 protein kinase (p70S6K) signaling pathway, as a central controller of cell growth, proliferation, survival, and differentiation in response to extracellular signals, growth factors, nutrient availability, energy status of the cell, and stress, has recently gained attention in neuroscience. The effects of this signaling pathway on repair of spinal cord injury (SCI), however, have not been well elucidated. ATP is increasingly recognized as an important regulator of signal transduction pathways, and plays important roles in functional recovery after nervous system injury. In the present study, we examined the ATP-induced changes of the Akt/mTOR/p70S6K signaling pathway in injured spinal cord of adult rats and potential therapeutic effects of this pathway on SCI-induced locomotor dysfunction. SCI was produced by extradural weight-drop using modified Allen's stall with damage energy of 50 g-cm force. The rats were divided into four groups: SCI plus ATP, SCI plus saline, SCI plus ATP and rapamycin, and sham-operated. Using immunostaining studies, Western blot analyses and real-time qualitative RT-PCR analyses, we demonstrated that the Akt/mTOR/p70S6K signaling pathway is present in the injured spinal cord and the expression of its components at the protein and mRNA levels is significantly elevated by exogenous administration of ATP following SCI. We observed the effectiveness of the activated Akt/mTOR/p70S6K signaling pathway in improving locomotor recovery, significantly increasing the expression of nestin, neuronal nuclei (NeuN), neuron specific enolase (NSE), and neurofilament 200 (NF200), and relatively inhibiting excessive reactive astrogliosis after SCI in a rapamycin-sensitive manner. We concluded that ATP injection produced a significant activation of the Akt/mTOR/p70S6K signaling pathway in the injured spinal cord and that enhancement of rapamycin-sensitive signaling produces beneficial effects on SCI-induced motor function defects and repair potential. We suggest that modulation of this protein kinase signaling pathway activity should be considered as a potential therapeutic strategy for SCI.

The alteration of Id-1 and TSP-1 expression in mucoepidermoid carcinoma associated with its clinical features and prognosis.

Expression of Id-1 (inhibitors of DNA binding/differentiation protein 1) and TSP-1 (thrombospondin-1) in mucoepidermoid carcinoma and their relationship to pathological features and prognosis was studied. Moderately and poorly differentiated groups had significantly higher Id-1 positive expression rate (p<0.05) than well differentiated carcinoma. Stages III-IV showed significant increase of Id-1 positive expression rate (p<0.05) compared with stages I and II. Id-1 positive expression was significantly higher in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After that, patients with negative Id-1 expression had significantly higher tumor-free survival than patients with positive expression (p<0.05). Correlation between the expression of Id-1 and TSP-1 in mucoepidermoid carcinoma was negative (p<0.05). Poorly differentiated groups show significantly lower TSP-1 positive expression rate than well differentiated groups (p<0.05). No significant differences of TSP-1 positive expression were detected with clinical stage. TSP-1 positive expression was significantly lower in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After 5 years, patients with positive TSP-1 expression had significantly higher tumor-free survival than patients with negative TSP-1. Positive Id-1 expression is associated with high malignancy/poor prognosis; positive TSP-1 expression is associated with low malignancy/good prognosis. Protein expression status may help assess tumor malignancy and patient prognosis.