Anti-mGluR5 encephalitis with mental disorders as the initial symptom： a case report / 四川精神卫生

This article reported the clinical features of a rare patient with anti-metabotropic glutamate receptor 5 （mGluR5） encephalitis with mental disorders as the initial symptom， so as to provide references for clinical diagnosis and treatment. The patient was a 38-year-old male， developed pharyngeal pain as prodromal symptoms， and the main clinical manifestations included rapidly progressive memory loss， anxiety and depression， and psychomotor excitement symptoms including irritability and impulsive behaviors. The disease had a progressive deterioration. In the most severe state， the patient became unconscious in a shallow coma， with further cognitive decline， hallucinations and delusions， and lack of self-awareness. Both cerebrospinal fluid and serum anti-mGluR5 antibody were strongly positive （1∶100）. After two sessions of hormone shock therapy， the patient showed significantly improvement in consciousness， cognitive， emotional and psychiatric dimensions.

Study of two kinds of Alzheimer's disease tree shrew models / 中国药理学通报

Aim To explore the optimized tree shrews model of Alzheimer's disease through comparison of the pathology changes of brain neurons between the two kinds of tree shrew models.Methods Fifty tree shrews were randomly divided into five groups with 10 in each group:control group,high dose D-galactose combined with ibotenic acid (IBO) group,low dose D-galactose combined with IBO group [intraperitoneal injection D-galactose combined with IBO injection into bilateral basal nucleus of Meynert (BNM)],high dose Aβ25-35 combined with IBO group,and low dose Aβ25-35 combined with IBO group (injection into bilateral BNM).Hematoxylin and eosin (HE) staining was used to observe the morphological changes of brain neurons.The expressions of choline acetyltransterase (ChAT) and synaptophysin(SYP) in the brains were detected by immunohistochemical staining.Western blot was used to detect the expression of amyloid beta 1-42 (Aβ1-42),amyloid precursor protein (APP) and phosphorylated tau protein (p-tau).Results The HE staining showed there were different degrees of morphological changes in the brains of model groups.The changes in the high dose D-galactose and high dose Aβ25-35 combined with IBO group were more obvious than those in low dose D-galactose and Aβ25-35 combined with IBO group.Immunohistochemical staining revealed that the levels of ChAT and SYP in the model groups decreased compared with control group,and the decline in high dose Aβ25-35 combined with IBO group was more marked than that in low dose Aβ25-35 combined with IBO group(P ＜0.01).Western blot revealed that the levels of Aβ1-42,APP,p-tau in the model groups increased compared with control group,and the rise in high dose Aβ25-35 combined with IBO group was more apparent than that in low dose Aβ25-35 combined with IBO group (P ＜ 0.05 or P ＜ 0.01).Conclusion The method of modeling by Aβ25-35 combined with IBO injection into bilateral BNM is more suitable for the establishment of Alzheimer's disease model.

Regional homogeneity in mild cognition impairment patients with amnestic and non-amnesic: a resting-state fMRI study / 中国神经精神疾病杂志

Objective To investigate regional spontaneous brain activity in mild cognition impairment (MCI) patients with amnesic (aMCI) and non-amnesic (naMCI).Methods Twenty-five aMCI patients,21 naMCI patients and 15 normal controls (NC) underwent resting-state functional magnetic resonance imaging.The regional homogeneity (ReHo) map of the whole brain was obtained by calculating the similarity of each voxel with its nearest 26 voxel time series.The differences of ReHo map across the whole brain among three groups were compared.Results In aMCI group,ReHo values were lower in right frontal lobe and higher in left middle temporal gyrus and left cerebellum compared with NC (P＜0.05,Alphasim correction).In naMCI group,ReHo values were higher in anterior cingulate cortex and right middle frontal gyrus and lower in right parahippocampa gyrus,right middle temporal gyrus as well as right precuneus compared with NC (P＜0.05,Alphasim correction).Compared with naMCI,the ReHo values were significantly higher in left prefrontal gyrus,left middle temporal gyrus and lower in right cerebellum (P＜0.05,Alphasim correction).Conclusion There are differences in spontaneous brain activity of left prefrontal gyrus,left middle temporal gyrus and right cerebellum between aMCI and naMCI,which may be used to differentiate brain function between aMCI and naMCI patients.

Preliminary establishment of integration of Alzheimer's disease and blood stasis syndrome tree shrew model and evaluation of intervention of Panax notoginseng saponins / 中国中药杂志

To establish the integration of Alzheimer's disease(AD) and blood stasis syndrome tree shrew model. Panax notoginseng saponins (PNS) was used to intervene the model to testify the stability of the model. The level of blood stasis of each group in the tree shrew model was evaluated by analyzing five traditional Chinese medicine(TCM) characterizations, four blood coagulation indexes, plasma nitric oxide (NO) level, plasma superoxide dismutase (SOD) level in each group. Hematoxylin and eosin(HE) staining was used to observe the morphological changes of brain hippocampal neuron cell of each group. Immunohistochemical staining was used to assay the ChAT and SYP levels in brain hippocampus of each group.The blood stasis characterization of the integration of disease and syndrome group was more obvious than the AD group, and that of the drug administration group was lower than that of the integration of disease and syndrome group. Aβ1-42, APP, P-Tau, ChAT and SYP level of AD group were lower than those in the blank group, which were further reduced in the model of integration of disease and syndrome. However, the administration of PNS relieved the reduction, indicating that the AD and blood stasis integration syndrome tree shrew model is stable.

Changing of the orbital volume after endoscopic transethmoid medial wall decompression combined with the endoscopic transethmoid intraconal fat-removal orbital decompression in thyroid associated ophthalmopathy / 中国内镜杂志

Objective To present the changing of the parameters of orbital volume after endoscopic transethmoid medial orbital wall decompression combined with the endoscopic transethmoid intraconal fat-removal orbital decompression in thyroid associated ophthalmopathy (TAO).Methods A retrospective chart was reviewed in 11 patients (20 eyes) receiving orbital decompression for the treatment of exophthalmos secondary to TAO from September 2014 to August 2015. All patients diagnosed TAO were in stable and inactive phase at least for 6 months. High-resolution computed tomography (HRCT) scan were performed in all patients before and 3-month after surgery. CT scan of orbit and computer-aided measurement software were used to measure the exophthalmos. Changing of the parameters of orbital volume were recorded for analysis its relationship with the amount of proptosis reduction.Results The exophthalmos was signiifcantly decreased after surgery. Medial rectus volume, fat volume and orbital volume were larger postoperatively. The changing of rectus volume and fat volume both had negative correlationship with the amount of proptosis.Conclusions The thickening of medial rectus 3-month postoperative may due to the removing of medial wall of orbit during the surgery. The stimulation of the surgery may be another reason. The changing of rectus volume and fat volume both had negative correlationship with the amount of proptosis.

Non-invasive treatment to grade 1 essential hypertension by percutaneous laser and electric pulse to acupoint with music: A randomized controlled trial / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study a non-drug therapy for hypertension disease by combining percutaneous laser and electric pulse stimulation to acupoint with music, and to test the efficiency of the combining treatment to grade 1 essential hypertension.</p><p><b>METHODS</b>A total of 174 patients with grade 1 essential hypertension were randomly assigned to 3 groups with a random number table after Chinese medicine (CM) syndrome differentiation: the photoelectric and musical treatment group (Group 1, with a self-developed multi-mode audio frequency pulse photoelectric therapeutic apparatus), acupuncture group (Group 2), and oral placebo group (Group 3), 58 cases per group. The curative effect of each group was evaluated by the changes of blood pressure and CM syndrome integral before and after treatment.</p><p><b>RESULTS</b>Compared with Group 3, there were significant decrease of blood pressure and CM syndrome integral in Group 1 and Group 2 (P<0.01). Compared with Group 2, Group 1 showed the highest decrease in systolic pressure (P<0.017). The total effective rate of anti-hypertension in Group 1 (91.38%, 53/58) was significantly higher than that in Group 2 (74.13%, 43/58) and Group 3 (18.97%, 11/58, P<0.05 or P<0.01); and that in Group 2 was also significantly higher than that in Group 3 (P<0.01). There were significant difference in the total effective rate of CM syndrome integral in both Group 1 (93.10%, 54/58) and Group 2 (84.48%, 49/58) as compared with Group 3 (17.24%, 10/58, P<0.01), while there was no significant difference between Group 1 and Group 2 (P>0.05).</p><p><b>CONCLUSIONS</b>The multi-mode audio frequency pulse photoelectric therapeutic apparatus, combining music, laser and electric pulse stimulation, is clinically useful for grade 1 essential hypertension. This "three in one" therapy method is non-invasive, easy and simple to handle. It is expected to be popularized as a new alternative treatment.</p>

The study of plasma levels of sCD40 and sCD40L in Alzheimer’s disease and mild cognitive impairment / 中国神经精神疾病杂志

Objective To explore the plasma levels of soluble CD40 (sCD40) and soluble CD40 ligand (sCD40L) in the patients with Alzheimer’s disease (AD) and those with amnestic mild cognitive impairment (aMCI). Methods The levels of plasma sCD40 and sCD40L were measured in 20 patients with AD, 35 patients with aMCI, and 32 cognitively normal controls (NC) using commercially available ELISAs. The cognitive function of AD and aMCI patients was mea?sured by mini-mental state examination (MMSE). Results There were significant differences in plasma sCD40 among AD, aMCI and NC groups (P<0.05) as the medians (the upper and lower quartiles) of plasma levels were 123.3 (97.4, 149.5) pg/mL, 102.9 (63.6, 124.0) pg/mL and 70.66 (51.0, 90.8) pg/mL, respectively. There were significant differences in plasma sCD40L among AD, aMCI and NC groups (P<0.05) as plasma levels were 537.0 (316.0, 1134.0) pg/mL, 316.0 (190.0,546.0) pg/mL and 167.0 (107.5,478.0) pg/mL. A negative correlation between the plasma concentrations of sCD40L and the MMSE scores was found in aMCI patients (r=-0.736, P<0.001). Conclusions There are relevant chang?es of plasma sCD40 and sCD40L levels in patients with AD and aMCI. The present results suggest that plasma levels of sCD40 and sCD40L may be appropriate biomarkers for AD patients and indicate that CD40-CD40L signaling may be in?volved in AD pathophysiology.

Clinical research on electrode and laser stimulating on acupoint combined with music therapy for grade 1 essential hypertension / 中国针灸

<p><b>OBJECTIVE</b>To explore safe, effective, simple and easy non-drug treatments for grade 1 essential hypertension.</p><p><b>METHODS</b>According to TCM syndrome differentiation, 126 cases of grade 1 essential hypertension were classified into 4 types: liver-fire hyperactivity syndrome, yin-deficiency and yang-hyperactivity syndrome, excessive phlegm-dampness syndrome, yin-yang deficiency syndrome, and then the patients were randomly divided into a photoelectric combined with musical treatment group (group A), an acupuncture group(group B) and a placebo group (group C). The acupoints were selected according to TCM syndrome differentiation in group A and group B, and multi-mode audio frequency pulse photoelectric therapeutic apparatus and acupuncture were used in the two groups respectively, once daily. Taichong (LR 3) and Quchi (LI 11) were selected in liver-fire hyperactivity syndrome, Taixi (KI 3) and Sanyinjiao (SP 6) were selected yi yin-deficiency and yang-hyperactivity syndrome, Zusanli (ST 36) and Fenglong (ST 40) were selected in excessive phlegm-dampness syndrome, while Taixi (KI 3) and Guanyuan (CV 4) were selected yi yirryang deficiency syndrome. The group C was treated with oral administration of starch tablet (25 mg), one tablet each time,three times everyday. Ten days were considered as one course, totally three courses were required in the three groups. The blood pressure and scores of TCM syndromes before and after treatment were compared among the three groups.</p><p><b>RESULTS</b>The blood pressure decreased significantly after treatment in group A and group B (all P<0.01), and the decrease in systolic blood pressure was more significant in group A (P < 0.05). The total effective rate was 90.5 / (38/42) in group A, which was superior to 71. 4 (30/42, P < 0.05) in group B and 19.1% (18/34, P<0. 01) in group C. The scores of TCM syndromes were both improved in group A and group B, but without significant difference between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>The clinical effect of multi-mode audio frequency pulse photoelectric therapeutic apparatus for treatment of grade 1 essential hypertension is reliable. Meanwhile, it has the advantages of a non-invasive and simple operation.</p>

Endoscopic trans-ethmoid medial orbital wall decompression combined with intraconal fat decompression for Graves' ophthalmopathy / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To present a new mini-invasive surgery for compressive optic neuropathy (CON) in Graves' ophthalmopathy (GO) by adequately decompressing the orbital apex and correcting proptosis, and to analyze its results.</p><p><b>METHODS</b>A retrospective chart was reviewed in 29 patients receiving orbital decompression for the treatment of CON secondary to GO from October 2006 to May 2011. All patients diagnosed CON were in stable and inactive phase of GO at least for 6 months. All patients received endoscopic transethmoid medial orbital wall decompression to reduce the compression on the orbital apex. In the meanwhile, an endoscopic transethmoid intraconal fat-removal orbital decompression was performed to remove parts of intraconal fat with a special aspiration/cutting instrument to further reduce the proptosis. All patients were followed up periodically.</p><p><b>RESULTS</b>of improvement of visual acuity (VA), color vision, and amount of proptosis reduction and incidence of induced diplopia 9 months after surgery was recorded for analysis its feasibility.</p><p><b>RESULTS</b>Forty-five orbits of 29 patients were included in the study. At the 9 months review, 44 of 45 eyes (97.8%) improved their VA from -0.65±0.30 (x±s) preoperatively to -0.24±0.22, with a mean improvement of 0.55±0.17 (t=-13.012, P<0.001), 23 of 29 eyes (79.3%) had improved color vision (P<0.001), and the mean reduction in proptosis was (7.07±1.59) mm (range 4-11 mm). Postoperative symmetry to within 2 mm were achieved in all patients. Except 1 patient complaining of deterioration in diplopia following surgery, no patients presented new on-set diplopia postoperatively.</p><p><b>CONCLUSION</b>The endoscopic transethmoid medial orbital wall decompression combined with the endoscopic transethmoid intraconal fat-removal orbital decompression is an effective treatment with minimal morbidity for both visional recovery and improvement of proptosis for CON in GO.</p>

Prognostic factors of 305 nasopharyngeal carcinoma patients treated with intensity-modulated radiotherapy / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Radiotherapy is effective in treating nasopharyngeal carcinoma (NPC). This study evaluated the treatment efficacy, toxicity, and prognostic factors of intensity-modulated radiotherapy (IMRT) in the treatment NPC.</p><p><b>METHODS</b>Between September 2003 and September 2006, 305 patients with NPC were treated with IMRT in Fujian Provincial Cancer Hospital. IMRT was delivered as follows: gross tumor volume (GTV) received 66.0-69.8 Gy in 30-33 fractions, high-risk clinical target volume (CTV-1) received 60.0-66.65 Gy, low-risk clinical target volume (CTV-2) and clinical target volume of cervical lymph node regions (CTV-N) received 54.0-55.8 Gy. Patients with stages III or IV disease also received cisplatin-based chemotherapy. All patients were assessed for local-regional control, survival, and toxicity.</p><p><b>RESULTS</b>With a median follow-up of 35 months (range, 5-61 months), there were 16, 8, and 39 patients who had developed local, regional, and distant recurrence, respectively. The 3-year rates of local control, regional control, metastasis-free survival, disease-free survival, and overall survival were 94.3%, 97.7%, 86.1%, 80.3%, and 89.1%, respectively. Multivariate analyses revealed that T-classification had no predictive value for local control and survival, whereas N-classification was a significant prognostic factor for overall survival (P < 0.001), metastasis-free survival (P < 0.001), and disease-free survival (P = 0.003). For stages III-IV disease, concurrent and adjuvant chemotherapy did not influence prognosis. The most severe acute toxicities included Grade III mucositis in 14 patients (4.6%), Grade III skin desquamation in 90 (29.5%), and Grades III-IV leucocytopenia in 20 (6.5%). There were 7% patients with Grade II xerostomia after 2 years of IMRT, no Grades 3 or 4 xerostomia was detected.</p><p><b>CONCLUSIONS</b>IMRT provided favorable locoregional control and survival rates for patients with NPC, even in those with locally advanced disease. The acute and late toxicities were acceptable. N-classification was the main factor of prognosis. Further study is needed on chemotherapy for patients with NPC.</p>

Research Progress on the Polyphasic Taxonomy of Acetic Acid Bacteria / 微生物学通报

Acetic acid bacteria are Gram-negative,obligate aerobic bacteria that have the ability to incompletely oxidize alcohols or sugars to organic acids as end products. The taxonomy of acetic acid bacteria has undergone many changes in the last 30 years. The early classification systems for these bacteria were based on morphological and biochemical characteristics. Today,the acetic acid bacteria are classified as the consensus result of a polyphasic analysis,combining phenotypic,chemotaxonomic and genotypic data. This paper reviewed the polyphasic taxonomy of acetic acid bacteria,mainly introduced the current classification of acetic acid bacteria,then discussed the application of phenotypic,chemotaxonomic and genotypic method in the taxonomy of acetic acid bacteria.

Isolation and Identification of BLN-2 Producing Poly-?-Glutamic Acid and Studies on Its Solid-state Fermentation / 微生物学通报

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.

Multi-center phase II clinical trial of humanized anti-epidermal factor receptor monoclonal antibody h-R3 combined with radiotherapy for locoregionally advanced nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safty of the humanized anti-epidermal factor receptor monoclonal antibody h-R3 in combination with radiotherapy for locoregionally advanced nasopharyngeal carcinoma.</p><p><b>METHODS</b>Totally, 137 patients from 7 medical center around China were randomly divided into combined therapy group or control group. There was no difference in Karnofsky performance score between two groups. All patients in both groups received radical conventionally fractionated radiotherapy to the total dose of D(T) 70-76 Gy. For the combined therapy group, h-R3 was added at a dose of 100 mg i.v. weekly for 8 weeks started at the beginning of radiotherapy.</p><p><b>RESULTS</b>Of the 137 eligilbe patients, 70 were in the combined therapy group treated by h-R3 plus radiotherapy and 67 in the control group by radiotherapy alone. The intent-to-treat (ITT) population consisted of 130 patients, while the per-protocol (PP) population was composed of 126 patients. The efficacy was assessed respectively at three point of time: the end of treatment, the 5th- and 17th-week after treatment. The complete response (CR) of the combined therapy group was significantly higher than that of the control group in both ITT and PP (ITT: 65.63%, 87.50%, 90.63% versus 27.27%, 42.42%, 51.52%; PP: 67.21%, 90.16%, 93.44% versus 27.69%, 43.08%, 52.31%; P < 0.05, respectively). The most common h-R3-related adverse reactions were fever (4.3%), hypotension (2.9%), nausea (1.4%), dizziness (2.9%) and rash (1.4%), which could be reversible if treated properly. Radiotherapy combined with 100 mg h-R3 i. v. weekly was tolerable and did not aggravate the side effects of radiation. The quality of life in the combined therapy group was comparable to that in the control group.</p><p><b>CONCLUSION</b>This phase 1 multicenter clinical trial shows that h-R3 in combination with radiotherapy is effective and well-tolerated for the treatment of locoregionally advanced nasopharyngeal carcinoma.</p>

Identification and Simulation Mutation of a High-productive Strain of Poly(?-glutamic acid) Independent of Glutamic Acid / 微生物学通报

A high productive poly ?-glutamic acid(?-PGA) strain PGA-N in a culture medium containing no L-glutamine was isolated from fermentation products.With the following identifications of colony mor-phology,physiological and biochemistry experiments,and genetics,the strain PGA-N was classified as a Bacillus licheniformis.According to the product environment,the base culture medium having no L-glutamine was simulated.In order to enhance the production of the strain PGA-N,the fermentation condi-tions,such as carbon source,nitrogen source,were optimized and the ?-glutamic acid production reached 5.16 g/L after getting the optimum formulation of this culture medium.PGA-N was mutagenized with com-bination of NTG and UV.A mutant PGA-N-C10 was screened which PGA production was increased from 5.16 g/L to 8.82 g/L.The study also investigated the effects of agitation speed on the cell biomass,?-PGA production and the ?-PGA molecular weight.The ?-PGA yield of PGA-N-C10 was as high as 11.00 g/L when the agitation speed was 400 r/min.

The Developments of Microbial Resistance to Industrial Antiseptics and Disinfectants / 微生物学通报

These years with industrial antiseptics and disinfectants widely used, especially unscientifically used, the resistance of microorganisms is more and more serious, which brought much more difficulties to industrial producing. The developments on the resistance of microorganisms to industrial antiseptics and disinfectants, their resistant mechanisms and the control strategies are reviewed, which aids us to reasonably use industrial organic antiseptics and disinfectants in existence and provides academic and scientific basis.

The Microorganism Constitutes Analysis of Soy Sauce Liquor from High-Salt-Level Watery State Fermentation / 微生物学通报

A system analysis about the microbial flora of normal and abnormal soybean sauce liquor from the high-salt-level watery state fermentation was made and the dominant bacteria and yeasts were identified.On the other hand,a discussion of effect of temperature on microbial flora was made.The results indicated that there were no obvious differences about the count of aerobe,spore-producing bacteria,enterobacteriaceae,lactic acid bacteria and anaerobe between the normal and abnormal soybean sauce liquor and there were marked differences about the count of yeasts and salt-tolerant bacteria.The predominant yeasts in normal soybean sauce were Torulopsis and Saccharomyces,accounting for 55.9% and 35.3% of the total yeasts separately,and in abnormal soybean sauce were Pichia,candida and Saccharomyces,accounting for 62.8%,17.9% and 9.0% respectively.

Screening and Identification of an Independent-glutamic Acid Strain Producing Poly (?-glutamic acid) / 微生物学通报

17 strains of bacterium that produced a large amount of ?-PGA when it was grown aerobically in a culture medium containing ammonium salt and sugar as sources of nitrogen and carbon respectively,were isolated from bean products.With the following identifications of colony morphology,physiological and biochemistry experiments,and genetics,the strain PGA-O-7 was classified as a Bacillus subtilis.The PGA production 2.8 (mg/mL) was obtained when it was grown in a medium containing 3% ammonium sulfate and 4% glucose at 30℃ for 72h with sharking.

The Concentration and Variation of Airborne Microbe in Guangzhou City / 微生物学通报

One year survey on the concentrations and monthly or seasonal variations of airborne microbe in Guangzhou city were analysed and studied with JWL-IIB airborne microbial sampler. The results showed that the yearly average airborne microbe content of outdoor was 2, 298 cfu/m3, and that of indoor was 1,792 cfu/m3 in Guangzhou city. The monthly variation range of outdoor airborne microbe was from 1,073 to 4,096 cfu/m3, the highest content was 4,096 cfu/m3 in March, and the lowest content was 1,073 cfu/m3 in October. The outdoor airborne bacteria and fungi counts were the highest in spring, next in summer, lower in winter and the lowest in autumn in the four seasons . The yearly average concentrations of outdoor airborne microbe at the Garbage compression station, the business walk street, the key traffic route, the residential area, the industrial district and the garden were 4, 573, 3, 835, 1, 580, 1,413, 1, 197 and 1, 187 cfu/m3respectively; and Ones of indoor at the key traffic route, the tourist three star-route hotel and the subway station were 2,511, 1,699and 1,167 cfu/m3 respectively . The study on airborne microbe can be used for the research of health prevention and environment control measures in Guangzhou .

Isolation and Identification of Osmotolerant Yeast from "Swollen Can" Soy Sauce / 微生物学通报

Three osmotolerant yeasts were isolated from three batches of "swollen can" soy sauce produced by a Guangdong condiment plant. These strains grew faster in the media containing 50%～60% glucose or 15% NaCl than in common yeast media. The three yeasts were identified as Pichia etchellsii by using morphological characteristics, physiological and biochemical tests.